Ribonucleotide reductase small subunit p53R2 facilitates p21 induction of G1 arrest under UV irradiation.
p53R2, which is one of the two known ribonucleotide reductase small subunits (the other being M2), is suggested to play an important role in supplying deoxynucleotide triphosphates (dNTP) for DNA repair during the G(1) or G(2) phase of the cell cycle. The ability of p53R2 to supply dNTPs for repairing DNA damages requires the presence of a functional p53 tumor suppressor. Here, we report in vivo physical interaction and colocalization of p53R2 and p21 before DNA damage. Mammalian two-hybrid assay further indicates that the amino acids 1 to 113 of p53R2 are critical for interacting with the NH(2)-terminal region (amino acids 1-93) of p21. The binding between p21 and p53R2 decreases inside the nucleus in response to UV, the time point of which corresponds to the increased binding of p21 with cyclin-dependent kinase-2 (Cdk2), and the decreased Cdk2 activity in the nucleus at G(1). Interestingly, p53R2 dissociates from p21 but facilitates the accumulation of p21 in the nucleus in response to UV. On the other hand, the ribonucleotide reductase activity increases at the corresponding time in response to UV. These data suggest a new function of p53R2 of cooperating with p21 during DNA repair at G(1) arrest.


[A phytosociological interpretation of vegetation from sandy hills of the Peruvian desert].
The vegetation of the sandy hills ("lomas") constitutes the main originality of the Peruvian and Chilean desert with a high number of endemics that shapes the vicarious associations. In this work, a phytosociological view of sandy environments of the Peruvian coastal desert is presented. According to the Braun-Blanquet method, we have made up 32 phytosociological inventories and added 138 ones from others authors. In each inventory, we have analyzed its floristic composition and ecological parameters, as altitude, soil and geomorphology. All releves were synthesized in a table to deduce the different associations, higher phytosociological units, and the distribu tion of its flora along the Peruvian coast and the Andean Cordillera. Using the Shannon-Wiener diversity index, the diversity of this flora is discussed making a comparison with historical data about the use of the territory with livestock during pre-Inca and Inca cultures, and Spanish invasion. As a result, two associations from Southern Peru -Nolanetum scaposo-spathulatae and Palauetum camanensis-weberbaueri-, two alliances -Nolanion humifusae from central Peru, and Nolanion spathulatae from the Southern Peru- and a new order -Tetragonio crystallinae-Plantaginetalia limensis- are described. In Nolanetum scaposo-spathulatae, Dictyophragnus englerianus, Leptoglossis lomana, Nolana scaposa, N. spathulata, Palaua velutina and Tetragonia vestita are the main characteristics, while in Palauetum camanensis-weberbaueri association N. scaposa and P. velutina are replaced by Palaua camanensis and P. weberbaueri. Nolanion humifusae alliance integrates species as Geranium limae, Hymenocallis amancaes, Nolana humifusa, N. latipes, Palaua rhombifolia or Villanova oppositifolia. Likewise, Cistanthe weberbaueri, Cryptantha parviflora, Hoffmannseggia miranda, Lupinus mollendoensis, Nolana confinis, N. pallidula, N. scaposa, N. spathulata, Palaua camanensis, P. velutina, P. weberbaueri, Tetragonia vestita and Weberbauerella brongniartioides are the characteristic species of Nolanion spathulatae alliance. The Tetragonio crystallinae-Plantaginetalia limensis order presents characteristic plants don't linked with eutrophic soils, as Calandrinia alba, Cryptantha limensis, Dyschoriste repens, Monnina macrostachya, Oxalis lomana, Palaua malvifolia, Pectocarya lateriflora, Plantago limensis or Tetragonia crystallina, with a distribution that claps the geographical area of the new alliances. On the other hand, the vegetation of the desert ravines is discussed in the context of the coastal river plant communities and its disturbance by the dunes. After the application of the Shannon-Wiener diversity index on the synthetic table columns, we can deduce that an increase in Andean and European ruderal species is linked to an intensive livestock activity. The transhumance between the Andes and the coast from pre-Inca times until now, produces the plant dispersion of high Andean plants toward the coast; the Spanish colonization was the origin of the presence of European plants in the "lomas" vegetation of Peru.


Multiple myeloma in the older adult: better prospects, more challenges.
Multiple myeloma (MM) is disproportionately diagnosed in older adults; with the aging of the population, the number of older adults diagnosed with MM will increase by nearly 80% in the next two decades. Duration of survival has improved dramatically over the last 20 years, but the improvements in older adults have not been as great as those in younger adults with MM. In this article, we address treatment approaches in older adults who are eligible for and those ineligible for high-dose therapy with autologous stem-cell transplantation as well as supportive care considerations and the potential role for geriatric assessment in facilitating decision making for older adults with MM. The evidence from recent studies demonstrates that combinations of novel and conventional antimyeloma agents result in improved response rates and, in some cases, improved progression-free and overall survival. However, some older adults are particularly vulnerable to toxicities of therapy and discontinuation of therapy and, consequently, they have poorer survival. In addition, older adults may prioritize other outcomes of therapy, such as quality of life, over more conventional end points such as disease response and duration of survival. Geriatric assessment can facilitate risk-stratification of older adults at greater risk for adverse events from therapy and aid in personalizing therapy for vulnerable or frail older adults. Survival in older adults with MM is improving with novel therapeutics, but efficacy must be balanced with risk of toxicity of therapy and maintenance of quality of life. Novel instruments such as geriatric assessment tools may facilitate these aims.


Self-rated health and interleukin-6: Longitudinal relationships in older adults.
Both self-rated health (SRH) and inflammation are implicated in chronic diseases and premature mortality. Better SRH is associated with lower proinflammatory cytokines, but there is little evidence about whether this relationship is more stable or dynamic. To study the between- and within-person associations between SRH and IL-6. Older adults (N=131; Mage=75 years) rated their health and provided blood samples for analysis of IL-6 at separate occasions every 6 months over a period up to 5 years. Age, sex, BMI, neuroticism, and statin use were examined as covariates in multilevel models. In bivariate models, better SRH, lower BMI, younger age, and female sex correlated with lower IL-6. In multilevel models, stable SRH (between-person differences; p<.001) but not dynamic SRH (within-person changes; p=.93) correlated with IL-6. The stable relationship persisted with demographic and health covariates in the model. Better stable SRH but not dynamic SRH was robustly associated with lower IL-6 among older adults, lending support to previous cross-sectional findings on the relation between inflammatory markers and SRH. The findings suggest that trait-like mechanisms, rather than changes over a time scale of 6-month waves, govern this association. To further investigate the mechanisms behind the SRH-IL-6 association, studies with different measurement frequencies, higher within-person variability, and experimental approaches are warranted.


Cancer biomarkers--an invitation to the table.
The allure of the emerging genomic technologies in cancer is their ability to generate new biomarkers that predict how individual cancer patients will respond to various treatments. However, productive implementation of cancer biomarkers into patient care will require fundamental changes in how we consider approvals for cancer indications and how we track patient responses. Here we briefly describe ongoing efforts to identify and to validate cancer biomarkers, discuss the technological hurdles that lie ahead, and then focus on the more pressing political and cultural issues that, if left unheeded, could derail many of the anticipated benefits of biomarker research.


A combination of molecular markers and clinical features improve the classification of pancreatic cysts.
The management of pancreatic cysts poses challenges to both patients and their physicians. We investigated whether a combination of molecular markers and clinical information could improve the classification of pancreatic cysts and management of patients. We performed a multi-center, retrospective study of 130 patients with resected pancreatic cystic neoplasms (12 serous cystadenomas, 10 solid pseudopapillary neoplasms, 12 mucinous cystic neoplasms, and 96 intraductal papillary mucinous neoplasms). Cyst fluid was analyzed to identify subtle mutations in genes known to be mutated in pancreatic cysts (BRAF, CDKN2A, CTNNB1, GNAS, KRAS, NRAS, PIK3CA, RNF43, SMAD4, TP53, and VHL); to identify loss of heterozygozity at CDKN2A, RNF43, SMAD4, TP53, and VHL tumor suppressor loci; and to identify aneuploidy. The analyses were performed using specialized technologies for implementing and interpreting massively parallel sequencing data acquisition. An algorithm was used to select markers that could classify cyst type and grade. The accuracy of the molecular markers was compared with that of clinical markers and a combination of molecular and clinical markers. We identified molecular markers and clinical features that classified cyst type with 90%-100% sensitivity and 92%-98% specificity. The molecular marker panel correctly identified 67 of the 74 patients who did not require surgery and could, therefore, reduce the number of unnecessary operations by 91%. We identified a panel of molecular markers and clinical features that show promise for the accurate classification of cystic neoplasms of the pancreas and identification of cysts that require surgery.


Pharmacologic characteristics of potentiation of 5-HT3 receptors by alcohols and diethyl ether in NCB-20 neuroblastoma cells.
We have examined the actions of alkanols, halogenated ethanol derivatives and diethyl ether on ion current mediated by 5-HT3 receptors in NCB-20 neuroblastoma cells. The alcohols and diethyl ether potentiated 5-HT3 receptor-mediated ion current at concentrations that had no effect on membrane current when applied in the absence of agonist. The potency of alcohols increased with increasing hydrophobicity. However, the maximal efficacy of alcohols was unrelated to hydrophobicity. Interactions between different drugs applied simultaneously to cells were examined to determine whether these compounds compete for a distinct modulatory site associated with the 5-HT3 receptor. Analysis of interactions observed at different drug concentrations indicated a variety of interactions between different compounds, ranging from negative to positive allosteric interactions. Interactions between trichloroethanol (TCEt) and isopentanol exhibited characteristics that might indicate competition for a single site of action. However, further examination of interactions between these two drugs indicated that although isopentanol altered the efficacy of co-applied TCEt, TCEt did not have a similar effect with respect to isopentanol. Furthermore, isopentanol did not alter the potency of TCEt for potentiation of receptor function. The absence of competitive interactions among alcohols indicates that a single "alcohol receptor" cannot be defined using established pharmacologic approaches. Our findings are most consistent with the idea that alcohols interact with several hydrophobic sites associated with the 5-HT3 receptor.


Predicting response originality through brain activity: An analysis of changes in EEG alpha power during the generation of alternative ideas.
Growing neurophysiological evidence points to a role of alpha oscillations in divergent thinking (DT). In particular, studies have shown a consistent EEG alpha synchronization during performance on the Alternative Uses Task (AUT), a well-established DT task. However, there is a need for investigating the brain dynamics underlying the production of a sequence of multiple, alternative ideas at the AUT and their relationship with idea originality. In twenty young adults, we investigated changes in alpha power during performance on a structured version of the AUT, requiring to ideate four alternative uses for conventional objects in distinct and sequentially balanced time periods. Data analysis followed a three-step approach, including behaviour aspects, physiology aspects, and their mutual relationship. At the behavioural level, we observed a typical serial order effect during DT production, with an increase of originality associated with an increase in ideational time and a decrease in response percentage over the four responses. This pattern was paralleled by a shift from alpha desynchronization to alpha synchronization across production of the four alternative ideas. Remarkably, alpha power changes were able to explain response originality, with a differential role of alpha power over different sensor sites. In particular, alpha synchronization over frontal, central, and temporal sites was able to predict the generation of original ideas in the first phases of the DT process, whereas alpha synchronization over centro-parietal sites persistently predicted response originality during the entire DT production. Moreover, a bilateral hemispheric effect in frontal sites and a left-lateralized effect in central, temporal, and parietal sensor sites emerged as predictors of the increase in response originality. These findings highlight the temporal dynamics of DT production across the generation of alternative ideas and support a partially distinct functional role of specific cortical areas during DT.


Determinants of Rejection Rate for Coronary CT Angiography Fractional Flow Reserve Analysis.
Background Coronary artery fractional flow reserve (FFR) derived from CT angiography (FFTCT) enables functional assessment of coronary stenosis. Prior clinical trials showed 13%-33% of coronary CT angiography studies had insufficient quality for quantitative analysis with FFRCT. Purpose To determine the rejection rate of FFRCT analysis and to determine factors associated with technically unsuccessful calculation of FFRCT. Materials and Methods Prospectively acquired coronary CT angiography scans submitted as part of the Assessing Diagnostic Value of Noninvasive FFRCT in Coronary Care (ADVANCE) registry (https://ClinicalTrials.gov: NCT02499679) and coronary CT angiography series submitted for clinical analysis were included. The primary outcome was the FFRCT rejection rate (defined as an inability to perform quantitative analysis with FFRCT). Factors that were associated with FFRCT rejection rate were assessed with multiple linear regression. Results In the ADVANCE registry, FFRCT rejection rate due to inadequate image quality was 2.9% (80 of 2778 patients; 95% confidence interval [CI]: 2.1%, 3.2%). In the 10 621 consecutive patients who underwent clinical analysis, the FFRCT rejection rate was 8.4% (n = 892; 95% CI: 6.2%, 7.2%; P < .001 vs the ADVANCE cohort). The main reason for the inability to perform FFRCT analysis was the presence of motion artifacts (63 of 80 [78%] and 729 of 892 [64%] in the ADVANCE and clinical cohorts, respectively). At multivariable analysis, section thickness in the ADVANCE (odds ratio [OR], 1.04; 95% CI: 1.001, 1.09; P = .045) and clinical (OR, 1.03; 95% CI: 1.02, 1.04; P < .001) cohorts and heart rate in the ADVANCE (OR, 1.05; 95% CI: 1.02, 1.08; P < .001) and clinical (OR, 1.06; 95% CI: 1.05, 1.07; P < .001) cohorts were independent predictors of rejection. Conclusion The rates for technically unsuccessful CT-derived fractional flow reserve in the ADVANCE registry and in a large clinical cohort were 2.9% and 8.4%, respectively. Thinner CT section thickness and lower patient heart rate may increase rates of completion of CT fractional flow reserve analysis. Published under a CC BY 4.0 license. Online supplemental material is available for this article. See also the editorial by Sakuma in this issue.


Race, biochemical disease recurrence, and prostate-specific antigen doubling time after radical prostatectomy: results from the SEARCH database.
Whether black men are at increased risk for biochemical disease recurrence after radical prostatectomy (RP) is debatable. Once black men have developed disease recurrence, it is unknown whether they have more aggressive disease than white men. To address this issue, the authors examined racial differences in pathologic features, time to disease recurrence, and prostate-specific antigen (PSA) doubling time (PSADT) among a cohort of patients treated with RP. The authors analyzed 953 white and 659 black men who were treated at 5 medical centers comprising the Shared Equal Access Regional Cancer Hospital (SEARCH) Database between 1988 and 2006. The association between race, adverse pathologic features, and biochemical disease recurrence was examined. Among those patients who developed disease recurrence, time to recurrence and PSADT were compared between the races. Black men were on average 2.1 years younger (P < .001) and had higher median preoperative PSA levels (7.6 ng/mL vs 7.0 ng/mL; P < .001), yet presented with a lower clinical stage of disease (T1: 62% vs 44%; P < .001) and similar biopsy Gleason scores (P = .59). After adjusting for multiple clinical characteristics, black men were found to be as likely as white men to have adverse pathologic features (Gleason score >or=7, positive surgical margins, and seminal vesicle invasion) in the RP specimen and were less likely to have extracapsular extension (P = .03). Black men were more likely to have a biochemical disease recurrence (hazards ratio [HR] of 1.28; 95% confidence interval [95% CI, 1.07-1.54 [P = .006]). This increased risk was reduced slightly after adjustment for multiple clinical and pathologic features, and no longer achieved statistical significance (HR of 1.19; 95% CI, 0.97-1.45 [P = .09]). Among men who developed disease recurrence, the median PSADT was found to be similar among black men (17.0 months) and white men (14.6 months) (P = .26). Despite presenting with earlier clinical stage and similar pathologic features at RP, black men were found to be at a slightly increased risk for biochemical disease recurrence. However, these recurrences appear to be no more aggressive than those found in white men.


An in vitro method for nephrotoxicity evaluation using HK-2 human kidney epithelial cells combined with biomarkers of nephrotoxicity.
The kidney is one of the major target organs for drug-induced toxicity. During drug development, the traditional markers of nephrotoxicity indicate only severe and late damage, which leads to high costs. The new biomarkers are needed for a more sensitive and reliable evaluation of nephrotoxicity, especially for the regulatory accepted and validated in vitro model. We developed an in vitro model based on the HK-2 cell using the biomarkers of nephrotoxicity as endpoints for the evaluation of nephrotoxicity. The predictive performance of the biomarkers including LDH, GGT, KIM-1, clusterin, CysC, NGAL, TIMP-1, GSTπ and osteopontin was evaluated with 22 well characterized compounds. The area under the curve (AUC) values of KIM-1, clusterin, CysC and osteopontin ranged between 0.79 and 0.84. The combination of clusterin, KIM-1 and/or osteopontin improved the AUC value (ranging between 0.88 and 0.95) compared to one biomarker. Taken together, these results suggest that the model based on the HK-2 cell using clusterin, osteopontin, CysC and KIM-1 as endpoints would allow the prediction of nephrotoxicity at early preclinical stages.


The interplay of Western diet and binge drinking on the onset, progression, and outlook of liver disease.
Non-alcoholic fatty liver disease and alcoholic liver disease, the two most prevalent liver diseases worldwide, share a common pathology but have largely been considered disparate diseases. Liver diseases are widely underestimated, but their prevalence is increasing worldwide. The Western diet (high-fat, high-sugar) and binge drinking (rapid consumption of alcohol in a short period of time) are two highly prevalent features of standard life in the United States, and both are linked to the development and progression of liver disease. Yet, few studies have been conducted to elucidate their potential interactions. Data shows binge drinking is on the rise in several age groups, and poor dietary trends continue to be prevalent. This review serves to summarize the sparse findings on the hepatic consequences of the combination of binge drinking and consuming a Western diet, while also drawing conclusions on potential future impacts. The data suggest the potential for a looming liver disease epidemic, indicating that more research on its progression as well as its prevention is needed on this critical topic.


Altered social behavior in mice carrying a cortical Foxp2 deletion.
Genetic disruptions of the forkhead box transcription factor FOXP2 in humans cause an autosomal-dominant speech and language disorder. While FOXP2 expression pattern are highly conserved, its role in specific brain areas for mammalian social behaviors remains largely unknown. Here we studied mice carrying a homozygous cortical Foxp2 deletion. The postnatal development and gross morphological architecture of mutant mice was indistinguishable from wildtype (WT) littermates. Unbiased behavioral profiling of adult mice revealed abnormalities in approach behavior towards conspecifics as well as in the reciprocal responses of WT interaction partners. Furthermore mutant mice showed alterations in acoustical parameters of ultrasonic vocalizations, which also differed in function of the social context. Cell type-specific gene expression profiling of cortical pyramidal neurons revealed aberrant regulation of genes involved in social behavior. In particular Foxp2 mutants showed the downregulation of Mint2 (Apba2), a gene involved in approach behavior in mice and autism spectrum disorder in humans. Taken together these data demonstrate that cortical Foxp2 is required for normal social behaviors in mice.


Parasite-associated growth enhancement in a fish-cestode system.
Parasites impose an energetic cost upon their hosts, yet, paradoxically instances have been reported in which infection is associated with enhanced, rather than diminished, host growth rates. Field studies of these parasite effects are problematic, since the pre-infection condition of the hosts is generally unknown. Here, we describe a laboratory experiment in which the growth rate and body condition of 76 laboratory-reared three-spined stickleback fishes were examined before, during and after each fish was fed the infective stage of the parasitic cestode Schistocephalus solidus. Twenty-one of these fishes went on to become infected by the cestode. Fishes were individually housed and provided with an abundant food supply to eliminate the potentially masking effects of variable competitive ability. Infection occurred independently of fish gender, size, body condition or pre-exposure growth rate. After exposure to the cestode, infected fishes grew faster (excluding parasite weight) and maintained a similar or better body condition compared with uninfected fishes, despite developing enlarged spleens. The accelerated growth could not be explained by reduced gonadal development. This result, one of few demonstrations of parasite-associated growth enhancement in fishes, is discussed with respect to other such parasite systems.


[Development of software for three-dimensional reconstruction and automatic quantification of intravascular ultrasound images. Initial experience].
Quantification of intravascular ultrasound (IVUS) images is essential in ischemic heart disease and interventional cardiology. Manual analysis is very slow and expensive. We describe an automated computerized method of analysis that requires only minimal initial input from a specialist. This study was carried out by interventional cardiologists and biomedical engineers working in close collaboration. We developed software in which it was necessary only to identify the media-adventitia boundary in a few images taken from the whole sequence. A three-dimensional reconstruction was then generated from each sequence, from which measurements of areas and volumes could be derived automatically. In total, 2300 randomly selected images from video sequences of 11 patients were analyzed. Results obtained using the proposed method differed only minimally from those obtained with the manual method: for vessel area measurements, the variability was 0.08 (0.07) (mean absolute error [standard deviation] normalized to the actual value; this corresponds to an error of 0.08 mm(2) per mm(2) of vessel area); for lumen area, 0.11 (0.11) (normalized), and for plaque volume, 0.5 (0.3) (normalized). Regions with severe lesions (<4 mm(2)) were correctly identified in more than 90% of cases. Specialist time needed for each reconstruction was 10 (8) minutes (vs 60 [10] minutes for manual analysis; P< .0001). The computerized method used dramatically reduced the time and effort needed for IVUS sequence analysis, and the automated measurements obtained were very promising.


Assessment of Safety and Efficacy of Safinamide as a Levodopa Adjunct in Patients With Parkinson Disease and Motor Fluctuations: A Randomized Clinical Trial.
Although levodopa remains the most effective oral pharmacotherapy for Parkinson disease (PD), its use is often limited by wearing off effect and dyskinesias. Management of such complications continues to be a significant challenge. To investigate the efficacy and safety of safinamide (an oral aminoamide derivative with dopaminergic and nondopaminergic actions) in levodopa-treated patients with motor fluctuations. From March 5, 2009, through February 23, 2012, patients from academic PD care centers were randomized (1:1 ratio) to receive double-blind adjunctive safinamide or placebo for 24 weeks. All patients had idiopathic PD with "off" time (time when medication effect has worn off and parkinsonian features, including bradykinesia and rigidity, return) of greater than 1.5 hours per day (excluding morning akinesia). Their pharmacotherapy included oral levodopa plus benserazide or carbidopa in a regimen that had been stable for 4 weeks or longer. During screening, each patient's regimen was optimized to minimize motor fluctuations. Study eligibility required that after 4 weeks of optimized treatment, the patients still have more than 1.5 hours per day of off time. Adverse events caused the premature study discontinuation of 12 individuals (4.4%) in the safinamide group and 10 individuals (3.6%) in the placebo group. Patients took safinamide or placebo as 1 tablet daily with breakfast. If no tolerability issues arose by day 14, the starting dose, 50 mg, was increased to 100 mg. The prespecified primary outcome was each treatment group's mean change from baseline to week 24 (or last "on" treatment value) in daily "on" time (relief of parkinsonian motor features) without troublesome dyskinesia, as assessed from diary data. At 119 centers, 549 patients were randomized (mean [SD] age, 61.9 [9.0] years; 334 male [60.8%] and 371 white [67.6%]): 274 to safinamide and 275 to placebo. Among them, 245 (89.4%) receiving safinamide and 241 (87.6%) receiving placebo completed the study. Mean (SD) change in daily on time without troublesome dyskinesia was +1.42 (2.80) hours for safinamide, from a baseline of 9.30 (2.41) hours, vs +0.57 (2.47) hours for placebo, from a baseline of 9.06 (2.50) hours (least-squares mean difference, 0.96 hour; 95% CI, 0.56-1.37 hours; P < .001, analysis of covariance). The most frequently reported adverse event was dyskinesia (in 40 [14.6%] vs 15 [5.5%] and as a severe event in 5 [1.8%] vs 1 [0.4%]). The outcomes of this trial support safinamide as an effective adjunct to levodopa in patients with PD and motor fluctuations to improve on time without troublesome dyskinesia and reduce wearing off. clinicaltrials.gov Identifier NCT00627640.


Does early introduction of solid feeding lead to early cessation of breastfeeding?
Mixed milk feeding increases the likelihood of breastfeeding cessation, but it is not known if solid feeding (SF) has the same effect. We have identified 10,407 infants breastfed for at least 8-10 weeks from three large U.K. studies (Avon Longitudinal Study of Parents and Children [ALSPAC; born 1990-1991], Southampton Woman's Survey [SWS; 1998-2008], and Infant Feeding Survey 2010 [IFS 2010]) to investigate the associations between early SF and breastfeeding cessation. In the earliest study (ALSPAC), 67% had started SF before the age of 4 months, but in the latest (IFS), only 23% had started before 4 months. Solid food introduction before 4 months was associated with stopping breastfeeding before 6 months in all three cohorts, with little effect of adjustment for maternal sociodemographic characteristics (Poisson regression, adjusted prevalence ratios: ALSPAC 1.55, [95% confidence interval 1.4, 1.8], SWS 1.13 [1.0, 1.3], IFS 1.10 [1.1, 1.3]). Using Cox regression, adjusted hazard ratios for breastfeeding cessation compared with SF after 5 months were 2.07 (1.8, 2.4) for SF before 4 and 1.51 (1.3, 1.8) at 4-5 months for ALSPAC and 1.25 (1.1, 1.5) and 1.15 (1.0, 1.3) for SWS. Earlier introduction of solids was associated with a shorter duration of breastfeeding, particularly in cohorts where earlier introduction of solids was the norm, with a dose-response relationship, which was not explained by background social characteristics. As mothers most commonly introduced solids in the month prior to the then recommended age, continuing to recommend deferring solids to the age of 6 months is important to support sustained breastfeeding.


Functional dissection of Reelin signaling by site-directed disruption of Disabled-1 adaptor binding to apolipoprotein E receptor 2: distinct roles in development and synaptic plasticity.
The Reelin signaling pathway controls neuronal positioning in human and mouse brain during development as well as modulation of long-term potentiation (LTP) and behavior in the adult. Reelin signals by binding to two transmembrane receptors, apolipoprotein E receptor 2 (Apoer2) and very-low-density lipoprotein receptor. After Reelin binds to the receptors, Disabled-1 (Dab1), an intracellular adaptor protein that binds to the cytoplasmic tails of the receptors, becomes phosphorylated on tyrosine residues, initiating a signaling cascade that includes activation of Src-family kinases and Akt. Here, we have created a line of mutant mice (Apoer2 EIG) in which the Apoer2 NFDNPVY motif has been altered to EIGNPVY to disrupt the Apoer2-Dab1 interaction to further study Reelin signaling in development and adult brain. Using primary neuronal cultures stimulated with recombinant Reelin, we find that normal Reelin signaling requires the wild-type NFDNPVY sequence and likely the interaction of Apoer2 with Dab1. Furthermore, examination of hippocampal, cortical, and cerebellar layering reveals that the NFDNPVY sequence of Apoer2 is indispensable for normal neuronal positioning during development of the brain. Adult Apoer2 EIG mice display severe abnormalities in LTP and behavior that are distinct from those observed for mice lacking Apoer2. In Apoer2 EIG slices, LTP degraded to baseline within 30 min, and this was prevented in the presence of Reelin. Together, these findings emphasize the complexity of Reelin signaling in the adult brain, which likely requires multiple adaptor protein interactions with the intracellular domain of Apoer2.


Sampling for global epidemic models and the topology of an international airport network.
Mathematical models that describe the global spread of infectious diseases such as influenza, severe acute respiratory syndrome (SARS), and tuberculosis (TB) often consider a sample of international airports as a network supporting disease spread. However, there is no consensus on how many cities should be selected or on how to select those cities. Using airport flight data that commercial airlines reported to the Official Airline Guide (OAG) in 2000, we have examined the network characteristics of network samples obtained under different selection rules. In addition, we have examined different size samples based on largest flight volume and largest metropolitan populations. We have shown that although the bias in network characteristics increases with the reduction of the sample size, a relatively small number of areas that includes the largest airports, the largest cities, the most-connected cities, and the most central cities is enough to describe the dynamics of the global spread of influenza. The analysis suggests that a relatively small number of cities (around 200 or 300 out of almost 3000) can capture enough network information to adequately describe the global spread of a disease such as influenza. Weak traffic flows between small airports can contribute to noise and mask other means of spread such as the ground transportation.


Randomized clinical trial of ischaemic preconditioning in major liver resection with intermittent Pringle manoeuvre.
Vascular inflow occlusion is effective in avoiding excessive blood loss during hepatic parenchymal transection but may cause ischaemic damage to the remnant liver. Intermittent portal triad clamping (IPTC) is superior to continuous hepatic pedicle clamping as it avoids severe ischaemia-reperfusion (IR) injury in the liver remnant. Ischaemic preconditioning (IPC) before continuous Pringle manoeuvre may protect against IR during major liver resection. This RCT assessed the impact of IPC in major liver resection with intermittent vascular inflow occlusion. Patients undergoing major liver resection with intermittent vascular inflow occlusion were randomized, during surgery, to receive IPC (10 min inflow occlusion followed by 10 min reperfusion) or no IPC (control group). Data analysis was on an intention-to-treat basis. The primary endpoint was serum alanine aminotransferase (ALT) level on the day after surgery. Eighty four patients were enrolled and randomized to IPC (n = 41) and no IPC (n = 43). The groups were comparable in terms of demographic data, preoperative American Society of Anesthesiologists grade and extent of liver resection. Intraoperative morbidity and postoperative outcomes were also similar. ALT levels on the day after operation were not decreased by IPC (mean(s.d.) 537·6(358·5) versus 525·0(400·6) units/ml in IPC and control group respectively; P = 0·881). Liver biochemistry tests in the week after operation showed the same pattern in both groups. IPC did not reduce liver damage in patients undergoing major liver resection with IPTC. NCT00908245 (http://www.clinicaltrials.gov).


Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine.
We have developed a procedure for simultaneous flow cytometric measurement of cellular DNA content and amount of BrdUrd incorporated into cellular DNA. Propidium iodide was used as a fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used to fluorescently label the bound anti-BrdUrd probe. Bivariate DNA/BrdUrd distributions measured for Chinese hamster ovary cells labeled for 30 min with BrdUrd clearly show the G1-and G2M-phase cells to have low BrdUrd-linked fluorescence and the S-phase cells to have high BrdUrd-linked fluorescence. Cell cycle traverse rates were estimated for Chinese hamster ovary cells from bivariate distributions measured for samples taken periodically after pulse labeling with BrdUrd. Bivariate DNA/BrdUrd distributions were also applied in the analysis of the response of C3H murine bone marrow cells to treatment in vivo with 1-beta-D-arabinofuranosylcytosine (araC). Bivariate distributions were measured for bone marrow cells taken from mice that were pulse labeled with BrdUrd at various times after treatment with araC. The resulting DNA/BrdUrd sequences show the kinetics of recovery from araC and allow discrimination of the araC sterilized cells.


Epigenetic alterations and an increased frequency of micronuclei in women with fibromyalgia.
Fibromyalgia (FM), characterized by chronic widespread pain, fatigue, and cognitive/mood disturbances, leads to reduced workplace productivity and increased healthcare expenses. To determine if acquired epigenetic/genetic changes are associated with FM, we compared the frequency of spontaneously occurring micronuclei (MN) and genome-wide methylation patterns in women with FM (n = 10) to those seen in comparably aged healthy controls (n = 42 (MN); n = 8 (methylation)). The mean (sd) MN frequency of women with FM (51.4 (21.9)) was significantly higher than that of controls (15.8 (8.5)) (χ (2) = 45.552; df = 1; P = 1.49 × 10(-11)). Significant differences (n = 69 sites) in methylation patterns were observed between cases and controls considering a 5% false discovery rate. The majority of differentially methylated (DM) sites (91%) were attributable to increased values in the women with FM. The DM sites included significant biological clusters involved in neuron differentiation/nervous system development, skeletal/organ system development, and chromatin compaction. Genes associated with DM sites whose function has particular relevance to FM included BDNF, NAT15, HDAC4, PRKCA, RTN1, and PRKG1. Results support the need for future research to further examine the potential role of epigenetic and acquired chromosomal alterations as a possible biological mechanism underlying FM.


Conductometric Soot Sensors: Internally Caused Thermophoresis as an Important Undesired Side Effect.
Particulate matter sensors are of interest for application in the exhaust of any combustion processes, especially for automotive aftertreatment systems. Conductometric soot sensors have been serialized recently. They comprise planar interdigital electrodes (IDE) on an insulating substrate. Between the IDEs, a voltage is applied. Soot deposition is accelerated by the resulting electric field due to electrophoresis. With increasing soot deposition, the conductance between the IDE increases. The timely derivative of the conductance can serve as a sensor signal, being a function of the deposition rate. An increasing voltage between the IDE would be useful for detecting low particle exhausts. In the present study, the influence of the applied voltage and the sensor temperature on the soot deposition is investigated. It turned out that the maximum voltage is limited, since the soot film is heated by the resulting current. An internally caused thermophoresis that reduces the rate of soot deposition on the substrate follows. It reduces both the linearity of the response and the sensitivity. These findings may be helpful for the further development of conductometric soot sensors for automotive exhausts, probably also to determine real driving emissions of particulate matter.


Regulation of synthesis and activity of the PLSTIRE protein (cyclin-dependent kinase 6 (cdk6)), a major cyclin D-associated cdk4 homologue in normal human T lymphocytes.
The PLSTIRE protein (cyclin-dependent kinase 6 (cdk6)), which shares extensive sequence homology (approximately 70%) with cdk4, was identified as the earliest inducible member of the cdk family of proteins in human T lymphocytes induced to proliferate in vitro by stimulation either with phorbol 12,13-dibutyrate and ionomycin (PDB/I) or PHA. The p40cdk6 protein was present in resting cells and increased amounts were detected 6 h after stimulation. It increased in amount throughout the first cell cycle but was present in reduced amounts at later times. Activity of the kinase, determined by in vitro phosphorylation of recombinant truncated retinoblastoma tumor suppressor gene (Rb) protein (p60Rb), paralleled p40cdk6 protein amounts. Cyclins D2 and D3 were the major cyclins associated with p40cdk6, with D2 predominating in early G1 phase. Both PDB and ionomycin were required for maximal accumulation of p40cdk6, but either agent alone stimulated some increase in amount and activity of the protein. p40cdk6 also increased in amount in cells activated in the presence of cyclosporin A or FK506, drugs that inhibit production of IL-2 and cell proliferation, suggesting that initial induction occurred independently of IL-2-mediated cell cycle progression. Furthermore, increased accumulation of p40cdk6 protein and activity occurred in cells rendered "competent" (responsive to IL-2) by a brief treatment with PDB/I. Thus, increased accumulation of the protein and its activity begin before IL-2/IL-2 receptor interaction, suggesting that the cdk6-cyclin D2 complex might be involved in acquisition of the competent state in human T lymphocytes.


Exploring resistance pathways for first-generation NS3/4A protease inhibitors boceprevir and telaprevir using Bayesian network learning.
Resistance-associated variants (RAVs) have been shown to influence treatment response to direct-acting antivirals (DAAs) and first generation NS3/4A protease inhibitors (PIs) in particular. Interpretation of hepatitis C virus (HCV) genotypic drug resistance remains a challenge, especially in patients who previously failed DAA therapy and need to be retreated with a second DAA based regimen. Bayesian network (BN) learning on HCV sequence data from patients treated with DAAs could provide insight in resistance pathways against PIs for HCV subtypes 1a and 1b, in a similar way as applied before for HIV. The publicly available 'Rega-BN' tool chain was developed to study associative analyses for various pathogens. Our first analysis, comparing sequences from PI-naïve and PI-experienced patients, determined that NS3 substitutions R155K and V36M arise with PI-exposure in HCV1a infected patients, and were defined as major and minor resistance-associated variants respectively. NS3 variant 174H was newly identified as potentially related to PI resistance. In a second analysis, NS3 sequences from PI-naïve patients who cleared the virus during PI therapy and from PI-naïve patients who failed PI therapy were compared, showing that NS3 baseline variant 67S predisposes to treatment-failure and variant 72I to treatment success. This approach has the potential to better characterize the role of more RAVs, if sufficient therapy annotated sequence data becomes available in curated public databases. In addition, polymorphisms present in baseline sequences that predispose patients to therapy failure can be identified using this approach.


Role of cysteine residues in the function of human UDP glucuronosyltransferase isoform 1A1 (UGT1A1).
Bilirubin glucuronidation, catalysed by UGT1A1 [UGT (UDP glucuronosyltransferase) isoform 1A1, EC 220.127.116.11], is critical for biliary elimination of bilirubin. UGT1A1 deficiency causes CN-1 (Crigler-Najjar syndrome type 1), which is characterized by potentially lethal unconjugated hyperbilirubinaemia. Nucleotide sequence analysis of UGT1A1 in two CN-1 patients revealed that patient A was homozygous for a nt 530 G-->A (where nt 530 G-->A means guanine to adenine transition at nucleotide 530) mutation, predicting a C177Y substitution, and patient B had a nt 466 T-->C mutation on one allele and a nt 1070 A-->G mutation on the other, predicting a C156R and a Q357R substitution respectively. All 11 cysteine residues of mature human UGT1A1 are highly conserved in other human UGT isoforms and in rat, mouse and Rhesus monkey UGT1A1, suggesting their functional importance. Expression of mutagenized UGT1A1 plasmids showed that substitution of any of the seven cysteine residues located within the endoplasmic reticulum cisternae (including those mutated in patients A and B) abolished UGT1A1 activity or markedly increased its apparent K(m) for bilirubin. Substitution of the three cysteine residues within the C-terminal cytosolic tail had minimal effect on basal UGT1A1 activity, but prevented UGT1A1 activation by UDP-GlcNAc. N-Ethylmaleimide did not inhibit UGT1A1 activity in native microsomes, but prevented UGT1A1 activation by UDP-GlcNAc and inhibited the activity in digitonin-permeabilized microsomes. Dithiothreitol did not affect UGT1A1 activity in human liver microsomes. Together, the results suggested that free thiol groups, but not disulphide bonding, of seven cysteine residues within the intracisternal region of human UGT1A1 are important for its catalytic activity, while cysteine residues in the cytosolic domain may be involved in its physiological activation by UDP-GlcNAc.


Primary and secondary variants of juvenile psychopathy differ in emotional processing.
Accumulating research suggests that psychopathy can be disaggregated into low-anxious primary and high-anxious secondary variants, and this research may be important for understanding antisocial youths with callous-unemotional traits. Using model-based cluster analysis, the present study disaggregated 165 serious male adolescent offenders (M age = 16) with high scores on the Youth Psychopathic Traits Inventory into primary and secondary variants based on the presence of anxiety. The results indicated that the secondary, high-anxious variant was more likely to show a history of abuse and scored higher on measures of emotional and attentional problems. On a picture version of the dot-probe task, the low-anxious primary variant was not engaged by emotionally distressing pictures, whereas the high-anxious secondary variant was more attentive to such stimuli (Cohen d = 0.71). Although the two groups differed as hypothesized from one another, neither differed significantly in their emotional processing from a nonpsychopathic control group of offending youth (n = 208). These results are consistent with the possibility that the two variants of psychopathy, both of which were high on callous-unemotional traits, may have different etiological pathways, with the primary being more related to a deficit in the processing of distress cues in others and the secondary being more related to histories of abuse and emotional problems.


Vestibular-Evoked Cerebral Potentials.
The human vestibular cortex has mostly been approached using functional magnetic resonance imaging and positron emission tomography combined with artificial stimulation of the vestibular receptors or nerve. Few studies have used electroencephalography and benefited from its high temporal resolution to describe the spatiotemporal dynamics of vestibular information processing from the first milliseconds following vestibular stimulation. Evoked potentials (EPs) are largely used to describe neural processing of other sensory signals, but they remain poorly developed and standardized in vestibular neuroscience and neuro-otology. Yet, vestibular EPs of brainstem, cerebellar, and cortical origin have been reported as early as the 1960s. This review article summarizes and compares results from studies that have used a large range of vestibular stimulation, including natural vestibular stimulation on rotating chairs and motion platforms, as well as artificial vestibular stimulation (e.g., sounds, impulsive acceleration stimulation, galvanic stimulation). These studies identified vestibular EPs with short latency (<20 ms), middle latency (from 20 to 50 ms), and late latency (>50 ms). Analysis of the generators (source analysis) of these responses offers new insights into the neuroimaging of the vestibular system. Generators were consistently found in the parieto-insular and temporo-parietal junction-the core of the vestibular cortex-as well as in the prefrontal and frontal areas, superior parietal, and temporal areas. We discuss the relevance of vestibular EPs for basic research and clinical neuroscience and highlight their limitations.


Preliminary descriptions of transient-evoked and distortion-product otoacoustic emissions from graduates of an intensive care nursery.
Transient-evoked (TEOAE) and distortion-product otoacoustic emissions (DPOAE) were measured in 51 graduates of an intensive care nursery and compared to data obtained from 80 normal-hearing children and adults. All infants had click-evoked auditory brainstem responses (ABR) at 30 dB nHL or less while the older subjects had pure-tone thresholds of 20 dB HL or less for octave frequencies from 250 to 8000 Hz. OAE data were collected using commercially available devices. All data were analyzed in terms of emission amplitude, emission-to-noise ratio, and response reproducibility as a function of frequency. DPOAEs were measured at three points per octave between f2 frequencies of approximately 500 and 8000 Hz. TEOAEs were elicited by clicks and were analyzed in both octave and 1/3-octave bands centered at frequencies from 500 to 4000 Hz, as well as in the broadband condition. In addition, stimulus amplitudes for the clicks used to elicit TEOAEs were analyzed within octave and 1/3-octave bands to determine whether any age-related differences in responses can be accounted for on the basis of stimulus differences. Both emission amplitude and noise amplitude were greater in neonates than adults, although there was variability across frequency. Emission-to-noise ratio and response reproducibility were more similar between groups. For TEOAEs, high-frequency emission-to-noise ratios were larger in neonates compared to older subjects, while the reverse was true in the lower frequencies. Less obvious frequency effects were observed for DPOAEs. These findings are discussed in relation to the potential use of OAEs as screening measures for neonatal hearing loss.


Safety and efficacy of pramipexole in early Parkinson disease. A randomized dose-ranging study. Parkinson Study Group.
Monotherapy with dopamine agonists may be useful in early Parkinson disease. To evaluate dose-response relationships for tolerability, safety, and efficacy of the synthetic dopamine agonist pramipexole. Multicenter, multidosage, parallel-group, double-blind, placebo-controlled, randomized clinical trial. University or academically based movement disorder clinics. A total of 264 patients with early Parkinson disease (PD) who were not requiring or receiving levodopa or other dopamine agonists were enrolled. Subjects were randomized to 1 of 5 treatment groups: pramipexole doses of 1.5 mg/d, 3.0 mg/d, 4.5 mg/d, and 6.0 mg/d, or matching placebo. A 6-week dosage escalation period was followed by a 4-week maintenance period and a 1-week period during which active treatment was withdrawn. The primary measure of tolerability was the proportion of subjects completing the study on the assigned treatment. The primary measure of efficacy was the change from baseline to 10 weeks in the total score on the Unified Parkinson's Disease Rating Scale (UPDRS). Pramipexole was generally safe and well tolerated in this 10-week study. The proportion of subjects completing the study on the originally assigned dosage was 98% for placebo and 81% for the 1.5-mg/d, 92% for the 3.0-mg/d, 78% for the 4.5-mg/d, and 67% for the 6.0-mg/d treatment groups. There was a trend toward increased frequency of adverse experiences, particularly somnolence, in the 6.0-mg/d group. After 10 weeks of treatment, pramipexole-treated subjects showed a 20% improvement in total UPDRS scores, with mean improvements in scores ranging from 5.9 to 7.0 units among active treatment groups, compared with 0.9 units for the placebo group (P<.005 for each comparison with placebo). There was also evidence that the treatment effects were more pronounced in subjects with worse UPDRS scores at baseline. Pramipexole is safe and effective as short-term monotherapy in patients with early PD who are not receiving levodopa. Further study is warranted to determine the long-term impact of pramipexole on the progression of disability in PD and its value in comparison with levodopa therapy and other dopamine agonists.


Voluntary consumption of ethanol in 15 inbred mouse strains.
To determine genetic differences in ethanol consumption, 15 commonly used inbred strains of mice were given ad libitum two-bottle choice between ethanol, 0.2% saccharin, or ethanol plus saccharin in one bottle versus tap water in the other bottle. Three different concentrations of ethanol were used: 3%, 6% and 10% (v/v). Of the 15 strains, the C57BL/6J, C57BR/cdJ and C57L/J strains showed the most consistent higher intake of ethanol either with or without 0.2% saccharin. In marked contrast, the DBA/1J and DBA/2J strains consistently showed the lowest intake. Consumption of 3% ethanol without saccharin was highly genetically correlated with saccharin consumption (r = 0.77), suggesting that low concentrations of ethanol may have a sweet taste that affects voluntary consumption. Most strains showed very different patterns of response to ethanol with or without saccharin. Three patterns of strain responses were identified. Some strains avoided higher concentrations of ethanol whether in water or saccharin; some appeared to be sensitive to the ability of saccharin to mask the odor of ethanol; and some may have reduced consumption only when ethanol concentrations were high enough to produce aversive postingestional effects. Whereas earlier studies generally attempted to explain strain differences in consumption by invoking a single mechanism, our results demonstrate that more than one mechanism is necessary to explain the preferential ethanol intake of all strains studied.


Comparing core stability and traditional trunk exercise on chronic low back pain patients using three functional lumbopelvic stability tests.
It is a matter of controversy whether core stability exercise is preferred to other types of exercise for chronic low back pain. Lumbopelvic stability is an important element in low back pain. No study was found using lumbopelvic stability tests in comparing core stability and other exercises. The single leg squat, dip test, and runner pose test appear to be suitable as tests for lumbopelvic stability. The aim of this study was to compare "core stability" and "traditional trunk exercise" using these tests and also the Oswestry disability questionnaire and pain intensity. Twenty-nine non-specific chronic low back pain subjects were alternately allocated in one of the two exercise groups. For both groups, a 16-sessions exercise program was provided. Before and after training: (1) video was recorded while subjects performed the tests; (2) Oswestry disability questionnaire was completed; and (3) pain intensity was measured by visual analogue scale. The test videos were scored by three physiotherapists. Statistical analysis revealed a significant improvement in stability test scores (p = 0.020 and p = 0.041) and reduction in disability (p < 0.001) and pain (p < 0.001) within each group. No significant difference was seen between two groups in the three outcomes p = 0.41, p = 0.14, and p = 0.72. Insignificant differences between the two groups may indicate either non-specificity of CSE to increase lumbopelvic stability or equal effectiveness of TTE and CSE on improving LPS. The non-significant differences may also be attributable to the lack of sensitivity of our tests to assess stability change in two groups after training given the relatively small sample size.


Neuromuscular block and the electroencephalogram during sevoflurane anaesthesia.
The effect of neuromuscular block on the anaesthetic depth of hypnosis is an elusive question. We simultaneously investigated the influence of neuromuscular block on the bispectral index, a measure of hypnosis during general anaesthesia, and on the electroencephalogram. Patients were anaesthetized with sevoflurane. Noxious tetanic electrical stimulation was applied on two occasions: before and after profound neuromuscular block achieved with rocuronium. Neuromuscular block significantly attenuated the effect from noxious stimulation on electroencephalogram power and synchrony in the gamma band (P<0.05), and the corresponding effect on bispectral index (P<0.02). These findings are probably due to the reduced arousing afferent input from paralysed muscles, and not to changes in the frontal electromyogram.


Radiocontrast-induced nephropathy is attenuated by autophagy through regulation of apoptosis and inflammation.
Radiocontrast-induced nephropathy (RCN) is the third most common cause of acute renal failure among inpatients. Although the number of patients undergoing exams using radiocontrast is increasing, little progress has been made for RCN treatment. The pathophysiology of RCN is known as tubular injury due to oxidative stress. As autophagy regulates cellular damage under stressful conditions, we investigated the role of autophagy in RCN. RCN was induced in male C57BL/6 J mice by intraperitoneal injection of iohexol, and 3-methyladenine (3-MA) was used as an autophagy inhibitor. Tubular injury caused by iohexol was also examined in vitro using rat tubular cells (NRK-52E). Increased autophagy after iohexol administration was demonstrated by the increase of light chain 3-II in the damaged kidney tubules both in vivo and in vitro. Serum creatinine and tubular injury were significantly increased at 24 h after iohexol treatment, as compared to control group. Further they worsened with autophagy inhibition by 3-MA. In vitro studies also demonstrated that decreased cell viability by iohexol was aggravated with 3-MA pretreatment. Malondialdehyde measured for oxidative stress was increased by iohexol, and it was accentuated by autophagy inhibition, which resulted in increase of cytochrome c Apoptosis, increased by iohexol treatment, was augmented with autophagy inhibition. Macrophage infiltration and increase of monocyte chemotactic protein-1 in kidneys were induced by iohexol, and it was aggravated with autophagy inhibition. This study showed that autophagy was involved with the pathophysiology of RCN, and the role of autophagy in modulation of apoptosis, oxidative stress, and inflammatory cell infiltration was supposed as mechanisms mitigating RCN.


Effect of a monocyte chemoattractant protein-1 synthesis inhibitor on fibroblasts from patients with carpal tunnel syndrome.
Carpal Tunnel Syndrome (CTS) is an idiopathic fibrotic disorder. Fibrosis in the subsynovial connective tissues (SSCT) of CTS and many other fibrotic diseases is mediated by Transforming growth factor β (TGF-β). Recently monocyte chemoattractant protein-1 (MCP-1) a cytokine involved in cellular recruitment has been suggested to regulate TGF-β activity. It is related to the onset of diseases which are caused by fibrosis, such as idiopathic pulmonary fibrosis, renal fibrosis, and systemic scleroderma. In this study, we evaluated the effect of the MCP-1 synthesis inhibitor, Bindarit, on primary cultures of fibroblasts from the SSCT of five CTS patients. Fibroblasts were treated with Bindarit (10 μM, 50 μM, 100 μM, or 300 μM). Responses to inhibitors were evaluated by regulation of CTS fibrosis-associated genes, fibrosis gene array and Smad luciferase reporter assay. We also assessed the combination effect of Bindarit and SD208, a TGF-β receptor type 1 inhibitor on TGF-β signaling. Collagen type III A1 (Col3), connective tissue growth factor (CTGF), and SERPINE1 expression were significantly down-regulated by Bindarit (300 μM) compared to vehicle control. In the fibrosis array, expression of inhibin beta E chain precursor (INHBE), beta actin (ACTB), endothelin 1 (EDN1) and hypoxanthine phosphoribosyltransferase 1 (HPRT1) were significantly down-regulated, and integrin beta-3 (ITGB3) was significantly up-regulated by Bindarit (300 μM). Smad signal transduction activation was significantly down-regulated by Bindarit (300 μM) and/or SD208 (1 μM) with TGF-β1 compared to vehicle control with TGF-β1. These results suggest that Bindarit in combination with SD208 may be beneficial as medical therapy for the SSCT fibrosis associated with CTS.


Fractal structure underlying patterns of premature heart beats.
Distinctive patterns of premature heart beats are frequently observed in clinical data, yet transitions from one pattern to another often appear to occur unpredictably. In this study we investigate the origins of this apparent lack of long-term order by processing the time intervals between successive R wave features of the electrocardiogram to obtain a symbolic description of all characteristic patterns of premature beats observed in a clinical data set. These characteristic patterns are found to be 1) associated with fixed ranges of heart rate over periods of many hours and 2) organized into an interwoven fractal structure that corresponds to the sequence of mode-locked states predicted by a nonlinear mathematical model. The apparent randomness of transitions from one pattern to another results from the normal fluctuations in heart rate which can sample many such patterns on a minute-by-minute basis.


Impact of tumor cell VEGF expression on the in vivo efficacy of vandetanib (ZACTIMA; ZD6474).
VEGF is the key player in tumor angiogenesis. In the current study, the impact of VEGF expression on the response of tumors to the VEGFR2 associated tyrosine kinase inhibitor vandetanib was evaluated. Human colon carcinoma (HT29) and murine squamous carcinoma (SCCVII) clonal cell lines expressing varying levels of VEGF were established and their response to vandetanib was assessed in tissue culture and as solid tumors. Vandetanib treatment had no effect on tumor cell clonogenic cell survival in vitro but doses >or=10 nM significantly reduced endothelial cell migration. In vivo, tumors derived from cell clones expressing high levels of VEGF displayed significantly enhanced angiogenesis and more aggressive growth. An intradermal angiogenesis assay was used to demonstrate that a 4-day treatment with vandetanib (50 mg/kg/day) was able to significantly inhibit blood vessel growth induced by both parental and high VEGF-expressing tumor cell clones. In the HT29 tumor model, treatment response to vandetanib (50 mg/kg/day, Monday-Friday for 2 weeks) was greatest in xenografts derived from the highest VEGF-expressing cell clones. A similar trend was noted in the SCCVII tumor model. The present findings indicate that vandetanib therapy effectively counteracted the aggressive feature of tumor growth resulting from VEGF over-expressing tumor cells and suggest that such tumors may be particularly well suited for anti-VEGF interventions.


Evaluation of secondary school teachers' knowledge about psychosis: a contribution to early detection.
We sought to evaluate secondary school teachers' knowledge about psychosis and their level of interest in this topic given the key role they may be able to play in the early detection of psychosis. A questionnaire survey of 268 secondary school teachers from eight secondary schools within the Azienda Sanitaria Locale Rome/E and Rome/A catchment areas (two of the five city health districts) in Rome. Teachers were asked to complete the Italian version Esperienza e Conoscenza delle Difficoltò Sociali ed Emotive dei Giovani of the Knowledge and Experience of Social Emotional Difficulties among Young people self-report questionnaire that investigates the diagnosis, age of onset, aetiology, prognosis and treatment of psychosis. They were also asked about their experience with pupils with possible psychosis. Most of teachers were able to recognize psychotic symptoms from a case vignette. Approximately 25% of the teachers had experienced a pupil with possible symptoms of psychosis. However, teachers displayed little awareness of psychiatric community services available for young people. Our study shows that teachers may play an important role in early detection and psychosis prevention strategies. The teachers also displayed a keen interest in gaining a deeper knowledge of early psychotic signs and in cooperating closely with a mental health specialist to obtain guidance and support when faced with serious mentally ill pupils.


Global Multilocus Sequence Type Analysis of Chlamydia trachomatis Strains from 16 Countries.
The Uppsala University Chlamydia trachomatis multilocus sequence type (MLST) database (http://mlstdb.bmc.uu.se) is based on five target regions (non-housekeeping genes) and the ompA gene. Each target has various numbers of alleles-hctB, 89; CT058, 51; CT144, 30; CT172, 38; and pbpB, 35-derived from 13 studies. Our aims were to perform an overall analysis of all C. trachomatis MLST sequence types (STs) in the database, examine STs with global spread, and evaluate the phylogenetic capability by using the five targets. A total of 415 STs were recognized from 2,089 specimens. The addition of 49 ompA gene variants created 459 profiles. ST variation and their geographical distribution were characterized using eBURST and minimum spanning tree analyses. There were 609 samples from men having sex with men (MSM), with 4 predominating STs detected in this group, comprising 63% of MSM cases. Four other STs predominated among 1,383 heterosexual cases comprising, 31% of this group. The diversity index in ocular trachoma cases was significantly lower than in sexually transmitted chlamydia infections. Predominating STs were identified in 12 available C. trachomatis whole genomes which were compared to 22 C. trachomatis full genomes without predominating STs. No specific gene in the 12 genomes with predominating STs could be linked to successful spread of certain STs. Phylogenetic analysis showed that MLST targets provide a tree similar to trees based on whole-genome analysis. The presented MLST scheme identified C. trachomatis strains with global spread. It provides a tool for epidemiological investigations and is useful for phylogenetic analyses.


Dietary isoflavones, urinary isoflavonoids, and risk of ischemic stroke in women.
Hormone therapy has been shown to increase risk of ischemic stroke in women. Plant-derived estrogens, particularly soy isoflavones, are known to have some estrogenic effects and have been marketed as natural alternatives to hormone therapy. Concerns have been raised about whether high isoflavone exposure may be related to ischemic stroke risk as well. We examined the dietary intake of isoflavones and the urinary excretion of isoflavonoids in relation to risk of ischemic stroke in women. A prospective cohort study was conducted in 66,832 Chinese women (aged 40-70 y) who had no cardiovascular disease or cancer at baseline. Usual dietary intakes were assessed via in-person interviews with the use of a validated food-frequency questionnaire. Incident strokes were ascertained during follow-up home visits and confirmed by medical records. We also conducted a nested case-control study in postmenopausal women who had never used hormone therapy, including 1422 incident ischemic stroke cases and 1422 controls individually matched by age, date and time of urine sample collection, time since last meal, and use of antibiotics. Urinary isoflavonoids were measured with the use of high-performance liquid chromatography coupled with mass spectrometry. During a mean follow-up of 10 y, 3110 incident ischemic strokes were verified. Dietary isoflavone intake was associated with increased risk of ischemic stroke; multivariable-adjusted HRs from lowest to highest quintiles were 1.00, 1.05, 1.10, 1.11, and 1.24, respectively (95% CI: 1.08, 1.42; P-trend = 0.002). In the case-control study, a similar positive association was observed for dietary isoflavones, but no significant associations were shown for the urinary isoflavonoid concentration [OR: 1.01 (95% CI: 0.77, 1.32) for comparison of extreme quintiles]. A habitually high intake of soy isoflavones may be associated with a modest but significant increase in risk of ischemic stroke in women. However, no association was shown for the urinary excretion of isoflavonoids.


Intranasal insulin as a treatment for Alzheimer's disease: a review of basic research and clinical evidence.
Research in animals and humans has associated Alzheimer's disease (AD) with decreased cerebrospinal fluid levels of insulin in combination with decreased insulin sensitivity (insulin resistance) in the brain. This phenomenon is accompanied by attenuated receptor expression of insulin and insulin-like growth factor, enhanced serine phosphorylation of insulin receptor substrate-1, and impaired transport of insulin across the blood-brain barrier. Moreover, clinical trials have demonstrated that intranasal insulin improves both memory performance and metabolic integrity of the brain in patients suffering from AD or its prodrome, mild cognitive impairment. These results, in conjunction with the finding that insulin mitigates hippocampal synapse vulnerability to beta amyloid, a peptide thought to be causative in the development of AD, provide a strong rationale for hypothesizing that pharmacological strategies bolstering brain insulin signaling, such as intranasal administration of insulin, could have significant potential in the treatment and prevention of AD. With this view in mind, the review at hand will present molecular mechanisms potentially underlying the memory-enhancing and neuroprotective effects of intranasal insulin. Then, we will discuss the results of intranasal insulin studies that have demonstrated that enhancing brain insulin signaling improves memory and learning processes in both cognitively healthy and impaired humans. Finally, we will provide an overview of neuroimaging studies indicating that disturbances in insulin metabolism--such as insulin resistance in obesity, type 2 diabetes and AD--and altered brain responses to insulin are linked to decreased cerebral volume and especially to hippocampal atrophy.


Cis-regulatory PLETHORA promoter elements directing root and nodule expression are conserved between Arabidopsis thaliana and Medicago truncatula.
Nodules are unique organs formed on roots of legumes by soil-borne bacteria, collectively known as rhizobium. Recently, we have shown that orthologs of the AINTEGUMENTA-like (AIL) AP2 transcription factors PLETHORA (PLT) 1 to 4, that redundantly regulate Arabidopsis thaliana root development are involved in root and nodule growth in Medicago truncatula. Hence, it is conceivable that rhizobium has co-opted these genes for nodule development. Whether this co-option requires the presence of specific cis-elements in the promoters and/or specialization of PLT protein function is not clear. Here, we analyzed the qualitative expression patterns of the Arabidopsis PLT1 to 4 promoters in Medicago roots and nodules and compared these with the described expression patterns of the Medicago PLT genes. Our studies reveal that the expression patterns of the investigated promoters and their Medicago orthologs are very similar, indicating that at least all cis-elements regulating spatial PLT expression are conserved among the Arabidopsis and Medicago PLT1 to 4 promoters.


Novel CBG Derivatives Can Reduce Inflammation, Pain and Obesity.
Interest in CBG (cannabigerol) has been growing in the past few years, due to its anti-inflammatory properties and other therapeutic benefits. Here we report the synthesis of three new CBG derivatives (HUM-223, HUM-233 and HUM-234) and show them to possess anti-inflammatory and analgesic properties. In addition, unlike CBG, HUM-234 also prevents obesity in mice fed a high-fat diet (HFD). The metabolic state of the treated mice on HFD is significantly better than that of vehicle-treated mice, and their liver slices show significantly less steatosis than untreated HFD or CBG-treated ones from HFD mice. We believe that HUM-223, HUM-233 and HUM-234 have the potential for development as novel drug candidates for the treatment of inflammatory conditions, and in the case of HUM-234, potentially for obesity where there is a huge unmet need.


Repression of CC16 by cigarette smoke (CS) exposure.
Club (Clara) Cell Secretory Protein (CCSP, or CC16) is produced mainly by non-ciliated airway epithelial cells including bronchiolar club cells and the change of its expression has been shown to associate with the progress and severity of Chronic Obstructive Pulmonary Disease (COPD). In an animal model, the lack of CC16 renders the animal susceptible to the tumorigenic effect of a major CS carcinogen. A recent population-based Tucson Epidemiological Study of Airway Obstructive Diseases (TESAOD) has indicated that the low serum CC16 concentration is closely linked with the smoke-related mortality, particularly that driven by the lung cancer. However, the study of CC16 expression in well-defined smoke exposure models has been lacking, and there is no experimental support for the potential causal link between CC16 and CS-induced pathophysiological changes in the lung. In the present study, we have found that airway CC16 expression was significantly repressed in COPD patients, in monkey CS exposure model, and in CS-induced mouse model of COPD. Additionally, the lack of CC16 exacerbated airway inflammation and alveolar loss in the mouse model. Therefore, CC16 may play an important protective role in CS-related diseases.


Curcumin blocks brain tumor formation.
Turmeric, an essential ingredient of culinary preparations of Southeast Asia, contains a major polyphenolic compound, named curcumin or diferuloylmethane, which eliminates cancer cells derived from a variety of peripheral tissues. Although in vitro experiments have addressed its anti-tumor property, no in vivo studies have explored its anti-cancer activity in the brain. Oral delivery of this food component has been less effective because of its low solubility in water.We show that a soluble formulation of curcumin crosses the blood–brain barrier but does not suppress normal brain cell viability. Furthermore, tail vein injection, or more effectively, intracerebral injection through a cannula, blocks brain tumor formation in mice that had already received an intracerebral bolus of mouse melanoma cells (B16F10).While exploring the mechanism of its action in vitro we observed that the solubilized curcumin causes activation of proapoptotic enzymes caspase 3/7 in human oligodendroglioma (HOG) and lung carcinoma (A549) cells, and mouse tumor cells N18(neuroblastoma), GL261 (glioma), and B16F10. A simultaneous decrease in cell viability is also revealed by MTT [3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide]assays. Further examination of the B16F10 cells showed that curcumin effectively suppresses Cyclin D1, P-NF-kB, BclXL, P-Akt, and VEGF, which explains its efficacy in blocking proliferation, survival, and invasion of the B16F10 cells in the brain. Taken together,solubilized curcumin effectively blocks brain tumor formation and also eliminates brain tumor cells. Therefore, judicious application of such injectable formulations of curcumin could be developed into a safe therapeutic strategy for treating brain tumors.


High frequency of lactose intolerance in a prehistoric hunter-gatherer population in northern Europe.
Genes and culture are believed to interact, but it has been difficult to find direct evidence for the process. One candidate example that has been put forward is lactase persistence in adulthood, i.e. the ability to continue digesting the milk sugar lactose after childhood, facilitating the consumption of raw milk. This genetic trait is believed to have evolved within a short time period and to be related with the emergence of sedentary agriculture. Here we investigate the frequency of an allele (-13910*T) associated with lactase persistence in a Neolithic Scandinavian population. From the 14 individuals originally examined, 10 yielded reliable results. We find that the T allele frequency was very low (5%) in this Middle Neolithic hunter-gatherer population, and that the frequency is dramatically different from the extant Swedish population (74%). We conclude that this difference in frequency could not have arisen by genetic drift and is either due to selection or, more likely, replacement of hunter-gatherer populations by sedentary agriculturalists.


Artificial Intelligence in Primary Health Care: Perceptions, Issues, and Challenges.
Artificial intelligence (AI) is heralded as an approach that might augment or substitute for the limited processing power of the human brain of primary health care (PHC) professionals. However, there are concerns that AI-mediated decisions may be hard to validate and challenge, or may result in rogue decisions. To form consensus about perceptions, issues, and challenges of AI in primary care. A three-round Delphi study was conducted. Round 1 explored experts' viewpoints on AI in PHC (n=20). Round 2 rated the appropriateness of statements arising from round one (n=12). The third round was an online panel discussion of findings (n=8) with the members of both the International Medical Informatics Association and the European Federation of Medical Informatics Primary Health Care Informatics Working Groups. PHC and informatics experts reported AI has potential to improve managerial and clinical decisions and processes, and this would be facilitated by common data standards. The respondents did not agree that AI applications should learn and adapt to clinician preferences or behaviour and they did not agree on the extent of AI potential for harm to patients. It was more difficult to assess the impact of AI-based applications on continuity and coordination of care. While the use of AI in medicine should enhance healthcare delivery, we need to ensure meticulous design and evaluation of AI applications. The primary care informatics community needs to be proactive and to guide the ethical and rigorous development of AI applications so that they will be safe and effective.


[Treatment of asthma--an analysis of clinical judgment among general practitioners].
The asthma patient's symptoms, use of drugs, and peak expiratory flow rates dictate treatment of asthma according to the guidelines. Our aim was to examine which information influenced the decisions of general practitioners. Out of 100 general practitioners attending a course on asthma management, 75 responded to several constructed patient cases on asthma treatment. They decided whether or not to give oral steroids in case of an acute exacerbation and judged the quality of pharmacological treatment. Age, symptoms, body temperature, peak expiratory flow measurement and the dose of beta-2 agonists and inhaled steroids varied from patient to patient. Their attitudes to the use of oral steroids and to the usefulness of measuring peak expiratory flow were evaluated. Among the doctors, 52% evaluated only one characteristic of the patients when deciding upon treatment with oral steroids. The use of drugs was evaluated by one quarter of the doctors when they judged the quality of treatment. Respectively, 81% and 73% were influenced by objective measurements in these decisions. The majority of doctors applied oral steroids and measured peak expiratory flow in asthmatics. This study indicates that general practitioners focus mainly on measured peak expiratory flow rate. The patient's symptoms ought to be given more consideration in the treatment of asthma. The patient's use of anti-asthmatics is emphasised in guidelines, but was of only limited importance to the doctors in this study.


The importance of a normal breathing pattern for an effective abdominal-hollowing maneuver in healthy people: an experimental study.
A normal breathing pattern while performing the abdominal-hollowing (AH) maneuver or spinal-stabilization exercise is essential for the success of rehabilitation programs and exercises. In previous studies, subjects were given standardized instructions to control the influence of respiration during the AH maneuver. However, the effect of breathing pattern on abdominal-muscle thickness during the AH maneuver has not been investigated. To compare abdominal-muscle thickness in subjects performing the AH maneuver under normal and abnormal breathing-pattern conditions and to investigate the effect of breathing pattern on the preferential contraction ratio (PCR) of the transverse abdominis. Comparative, repeated-measures experimental study. University research laboratory. 16 healthy subjects (8 male, 8 female) from a university population. A real-time ultrasound scanner was used to measure abdominal-muscle thickness during normal and abnormal breathing patterns. A paired t test was used to assess the effect of breathing pattern on abdominal-muscle thickness and PCR. Muscle thickness in the transverse abdominis and internal oblique muscles was significantly greater under the normal breathing pattern than under the abnormal pattern (P < .05). The PCR of the transverse abdominis was significantly higher under the normal breathing pattern compared with the abnormal pattern (P < .05). The results indicate that a normal breathing pattern is essential for performance of an effective AH maneuver. Thus, clinicians should ensure that patients adopt a normal breathing pattern before performing the AH maneuver and monitor transverse abdominis activation during the maneuver.


Evaluation of the focal distance of lenses by white-light Lau phase interferometry.
We used a white-light Lau phase interferometer to evaluate the focal distances of two lenses. We found that the variation in the experimentally measured value is less than +/-1% from the given values. Limitations of the method and error analysis are presented.


The implications of megatrends in information and communication technology and transportation for changes in global physical activity.
Physical inactivity accounts for more than 3 million deaths per year, most from non-communicable diseases in low-income and middle-income countries. We used reviews of physical activity interventions and a simulation model to examine how megatrends in information and communication technology and transportation directly and indirectly affect levels of physical activity across countries of low, middle, and high income. The model suggested that the direct and potentiating eff ects of information and communication technology, especially mobile phones, are nearly equal in magnitude to the mean eff ects of planned physical activity interventions. The greatest potential to increase population physical activity might thus be in creation of synergistic policies in sectors outside health including communication and transportation. However, there remains a glaring mismatch between where studies on physical activity interventions are undertaken and where the potential lies in low-income and middle-income countries for population-level effects that will truly affect global health.


Arginine vasopressin concentrations in the cerebrospinal fluid of children.
Cerebrospinal fluid (CSF) arginine vasopressin (AVP) levels are reported in a group of 22 children (median age 24 months) investigated for possible bacterial meningitis and subsequently found not to be suffering from this disease. The mean CSF AVP concentration was 0.80 +/- 0.33 pg/ml. The results obtained in patients suffering from febrile convulsions (mean 0.71 pg/ml), other convulsive disorders (mean 0.80 pg/ml) and miscellaneous infectious diseases (mean 0.85 pg/ml) did not differ significantly from one another. Our findings confirm the presence of AVP in the CSF of children and provide reference values for further investigations into the functions of CSF AVP in children.


Graphene: a partially ordered non-periodic solid.
Molecular dynamics simulations were performed to study the structural features of graphene over a wide range of temperatures from 50 to 4000 K using the PPBE-G potential [D. Wei, Y. Song, and F. Wang, J. Chem. Phys. 134, 184704 (2011)]. This potential was developed by force matching the Perdew-Burke-Ernzerhof (PBE) exchange correlation functional and has been validated previously to provide accurate potential energy surface for graphene at temperatures as high as 3000 K. Simulations with the PPBE‑G potential are the best available approximation to a direct Car-Parrinello Molecular Dynamics study of graphene. One advantage of the PBE-G potential is to allow large simulation boxes to be modeled efficiently so that properties showing strong finite size effects can be studied. Our simulation box contains more than 600,000 C atoms and is one of the largest graphene boxes ever modeled. With the PPBE-G potential, the thermal-expansion coefficient is negative up to 4000 K. With a large box and an accurate potential, the critical exponent for the scaling properties associated with the normal-normal and height-height correlation functions was confirmed to be 0.85. This exponent remains constant up to 4000 K suggesting graphene to be in the deeply cooled regime even close to the experimental melting temperature. The reduced peak heights in the radial distribution function of graphene show an inverse power law dependence to distance, which indicates that a macroscopic graphene sheet will lose long-range crystalline order as predicted by the Mermin-Wagner instability. Although graphene loses long-range translational order, it retains long range orientational order as indicated by its orientational correlation function; graphene is thus partially ordered but not periodic.


Outcomes of Cardiac Catheterization in Patients With Atrial Fibrillation on Anticoagulation in Contemporary in Practice: An Analysis of the ORBIT II Registry.
Patients with atrial fibrillation on oral anticoagulation (OAC) undergoing cardiac catheterization face risks for embolic and bleeding events, yet information on strategies to mitigate these risks in contemporary practice is lacking. We aimed to describe the clinical/procedural characteristics of a contemporary cohort of patients with atrial fibrillation on OAC who underwent cardiac catheterization. Use of bleeding avoidance strategies and bridging therapy were described and outcomes including death, stroke, and major bleeding at 30 days and 1 year were compared by OAC type. Of 13 404 patients in the Outcomes Registry for Better Informed Treatment of Atrial Fibrillation II Registry from 2013 to 2016, 741 underwent cardiac catheterization (139 with percutaneous coronary intervention) in the setting of OAC. The patients' median age was 71, 61.8% were male, white (87.2%), had hypertension (83.7%), hyperlipidemia (72.1%), diabetes mellitus (31.6%), and chronic kidney disease (28.2%); 20.2% received warfarin while 79.8% received direct acting oral anticoagulant. One third of patients underwent radial artery access, and bivalirudin was used in 4.6%. Bridging therapy was used more often in patients on warfarin versus direct acting oral anticoagulant (16.7% versus10.0%). OAC was interrupted in 93.8% of patients. Patients on warfarin versus direct acting oral anticoagulant were equally likely to restart OAC (58.0% versus 60.7%), had similar use of antiplatelet therapy (44.0% versus 41.3%) after catheterization, and had similar rates of myocardial infarction and death at 1 year, but higher rates of major bleeding (43.3 versus 12.9 events/100 patient years) and stroke (4.9 versus 1.9 events/100 patient years). In a real-world registry of patients with atrial fibrillation undergoing cardiac catheterization, most cases are elective, performed by femoral access, with interruption of OAC. Bleeding avoidance strategies such as radial artery access and bivalirudin were used infrequently and use of bridging therapy was uncommon. Nearly 40% of patients did not restart OAC postprocedure, exposing patients to risk for stroke. Further research is necessary to optimize the management of patients with atrial fibrillation undergoing cardiac catheterization.


Cell-based vaccines for renal cell carcinoma: genetically-engineered tumor cells and monocyte-derived dendritic cells.
Initial vaccine developments for renal cell carcinoma (RCC) have concentrated on cell-based approaches in which tumor cells themselves provide mixtures of unknown tumor-associated antigens as immunizing agents. Antigens derived from autologous tumors can direct responses to molecular composites characteristic of individual tumors, whereas antigens derived from allogeneic tumor cells must be commonly shared by RCC. Three types of cell-based vaccine for RCC have been investigated: isolated tumor cell suspensions, gene modified tumor cells and dendritic cells (DCs) expressing RCC-associated antigens. Approaches using genetic modification of autologous RCC have included ex vivo modification of tumor cells or modification of tumors in vivo. We have used gene-modification of allogeneic tumor cell lines to create generic RCC vaccines. More recently, emphasis has shifted to the use of DCs as cell-based vaccines for RCC. DCs have moved to a position of central interest because of their excellent stimulatory capacity, combined with their ability to process and present antigens to both naive CD4 and CD8 cells. The long impasse in identifying molecular targets for specific immunotherapy of RCC is now rapidly being overcome through the use of tools and information emerging from human genome research. Identification of candidate molecules expressed by RCC using cDNA arrays, combined with protein arrays and identification of peptides presented by MHC molecules, allow specific vaccines to be tailored to the antigenic profile of individual tumors, providing the basis for development of patient-specific vaccines.


Ursolic acid and rosiglitazone combination improves insulin sensitivity by increasing the skeletal muscle insulin-stimulated IRS-1 tyrosine phosphorylation in high-fat diet-fed C57BL/6J mice.
The aim of this present study was to investigate the effect of ursolic acid (UA) and rosiglitazone (RSG) on insulin sensitivity and proximal insulin signaling pathways in high-fat diet (HFD)-fed C57/BL/6J mice. Male C57BL/6J mice were fed either normal diet or HFD for 10 weeks, after which animals in each dietary group were divided into the following six groups (normal diet, normal diet plus UA and RSG, HFD alone, HFD plus UA, HFD plus RSG, and HFD plus UA and RSG) for the next 5 weeks. UA (5 mg/kg BW) and RSG (4 mg/kg BW) were administered as suspensions directly into the stomach using a gastric tube. The HFD diet elevated fasting plasma glucose, insulin, and homeostasis model assessment index. The expression of insulin receptor substrate (IRS)-1, phosphoinositide 3-kinase (PI3-kinase), Akt, and glucose transporter (GLUT) 4 were determined by Western blot analyses. The results demonstrated that combination treatment (UA/RSG) ameliorated HFD-induced glucose intolerance and insulin resistance by improving the homeostatic model assessment (HOMA) index. Further, combination treatment (UA/RSG) stimulated the IRS-1, PI3-kinase, Akt, and GLUT 4 translocation. These results strongly suggest that combination treatment (UA/RSG) activates IRS-PI3-kinase-Akt-dependent signaling pathways to induce GLUT 4 translocation and increases the expression of insulin receptor to improve glucose intolerance.


Discrimination ratio analysis of inflammatory markers: implications for the study of inflammation in chronic disease.
To understand the role of inflammation in chronic disease it is important to have a reliable measure of habitual inflammatory status. A number of acute-phase response markers have been used as measures of inflammatory status, but the ability of a single measure to appropriately reflect habitual inflammatory status has not been assessed. This study compares the ability of different inflammatory markers to characterize habitual inflammatory status in overweight women. A single fasting blood sample was taken from 86 overweight women (mean body mass index [BMI], 35.2 kg/m2; range, 26.2 to 47.6 kg/m2) and a number of inflammatory markers (both acute-phase response markers and cytokines) were measured. A randomly selected subpopulation of 15 women attended on 2 further occasions for further blood samples. Using the subpopulation, discrimination ratios (DRs) were calculated for each inflammatory marker to assess the within-subject variability. The DRs were then used to determine the relationship between these markers, adjusted for within-subject variability, in the whole population. In this highly controlled experimental environment, interleukin-6 (IL-6), with a DR of 3.71, was the cytokine with the greatest ability to discriminate between subjects, suggesting that it is best able to characterize habitual inflammatory status. Sialic acid was the acute-phase response marker with the highest DR (3.16), and showed stronger correlations with other inflammatory markers, including C-reactive protein (CRP), than IL-6. This study suggests that use of some inflammatory markers, such as CRP, with large within-individual variability, will underestimate the relationship between inflammation and disease, and thus relationships between inflammation and chronic disease may be stronger than previously appreciated. Future studies should consider IL-6 or sialic acid to provide a more robust measure of inflammatory status.


HSP70 and other possible heat shock or oxidative stress proteins are induced in skeletal muscle, heart, and liver during exercise.
Exercise causes heat shock (muscle temperatures of up to 45 degrees C, core temperatures of up to 44 degrees C) and oxidative stress (generation of O2- and H2O2), and exercise training promotes mitochondrial biogenesis (2-3-fold increases in muscle mitochondria). The concentrations of at least 15 possible heat shock or oxidative stress proteins (including one with a molecular weight of 70 kDa) were increased, in skeletal muscle, heart, and liver, by exercise. Soleus, plantaris, and extensor digitorum longus (EDL) muscles exhibited differential protein synthetic responses ([3H]leucine incorporation) to heat shock and oxidative stress in vitro but five proteins (particularly a 70 kDa protein and a 106 kDa protein) were common to both stresses. HSP70 mRNA levels were next analyzed by Northern transfer, using a [32P]-labeled HSP70 cDNA probe. HSP70 mRNA levels were increased, in skeletal and cardiac muscle, by exercise and by both heat shock and oxidative stress. Skeletal muscle HSP70 mRNA levels peaked 30-60 min following exercise, and appeared to decline slowly towards control levels by 6 h postexercise. Two distinct HSP70 mRNA species were observed in cardiac muscle; a 2.3 kb mRNA which returned to control levels within 2-3 h postexercise, and a 3.5 kb mRNA species which remained at elevated concentrations for some 6 h postexercise. The induction of HSP70 appears to be a physiological response to the heat shock and oxidative stress of exercise. Exercise hyperthermia may actually cause oxidative stress since we also found that muscle mitochondria undergo progressive uncoupling and increased O2- generation with increasing temperatures.(ABSTRACT TRUNCATED AT 250 WORDS)


Potential of pharmacists to help reduce the burden of poorly managed cardiovascular risk.
Rural areas require better use of existing health professionals to ensure capacity to deliver improved cardiovascular outcomes. Community pharmacists (CPs) are accessible to most communities and can potentially undertake expanded roles in prevention of cardiovascular disease (CVD). This study aims to establish frequency of contact with general practitioners (GPs) and CPs by patients at high risk of CVD or with inadequately controlled CVD risk factors. Population survey using randomly selected individuals from the Wimmera region electoral roll and incorporating a physical health check and self-administered health questionnaire. Overall, 1500 were invited to participate. The participation rate was 51% when ineligible individuals were excluded. Nine out of 10 participants visited one or both types of practitioner in the previous 12 months. Substantially more participants visited GPs compared with CPs (88.5% versus 66.8%). With the exception of excess alcohol intake, the median number of opportunities to intervene for every inadequately controlled CVD risk factor and among high risk patient groups at least doubled for the professions combined when compared with GP visits alone. Opportunities exist to intervene more frequently with target groups by engaging CPs more effectively but would require a significant attitude shift towards CPs. Mechanisms for greater pharmacist integration into primary care teams should be investigated.


Transitions in Tobacco Product Use by U.S. Adults between 2013⁻2014 and 2014⁻2015: Findings from the PATH Study Wave 1 and Wave 2.
In 2013⁻2014, nearly 28% of adults in the United States (U.S.) were current tobacco users with cigarettes the most common product used and with nearly 40% of tobacco users using two or more tobacco products. We describe overall change in prevalence of tobacco product use and within-person transitions in tobacco product use in the U.S. between 2013⁻2014 and 2014⁻2015 for young adults (18⁻24 years) and older adults (25+ years). Data from Wave 1 (W1, 2013⁻2014) and Wave 2 (W2, 2014⁻2015) of the Population Assessment of Tobacco and Health (PATH) Study were analyzed (N = 34,235). Tobacco product types were categorized into: (1) combustible (cigarettes, cigars, pipe tobacco, hookah), (2) noncombustible (smokeless tobacco, snus pouches, dissolvable tobacco), and (3) electronic nicotine delivery systems (ENDS). Transitions for individual combustible-product types, and for single- and multiple-product use, were also considered. Overall prevalence of current tobacco use decreased from 27.6% to 26.3%. Among W1 non-tobacco users, 88.7% of young adults and 95.8% of older adults were non-tobacco users at W2. Among W1 tobacco users, 71.7% of young adults transitioned, with 20.7% discontinuing use completely, and 45.9% of older adults transitioned, with 12.5% discontinuing use completely. Continuing with/transitioning toward combustible product(s), particularly cigarettes, was more common than continuing with/transitioning toward ENDS. Tobacco use behaviors were less stable among young adults than older adults, likely reflecting greater product experimentation among young adults. Relative stability of cigarette use compared to other tobacco products (except older adult noncombustible use) demonstrates high abuse liability for cigarettes.