IKBKE activity enhances AR levels in advanced prostate cancer via modulation of the Hippo pathway.
Resistance to androgen receptor (AR) targeting therapeutics in prostate cancer (PC) is a significant clinical problem. Mechanisms by which this is accomplished include AR amplification and expression of AR splice variants, demonstrating that AR remains a key therapeutic target in advanced disease. For the first time we show that IKBKE drives AR signalling in advanced PC. Significant inhibition of AR regulated gene expression was observed upon siRNA-mediated IKBKE depletion or pharmacological inhibition due to inhibited AR gene expression in multiple cell line models including a LNCaP derivative cell line resistant to the anti-androgen, enzalutamide (LNCaP-EnzR). Phenotypically, this resulted in significant inhibition of proliferation, migration and colony forming ability suggesting that targeting IKBKE could circumvent resistance to AR targeting therapies. Indeed, pharmacological inhibition in the CWR22Rv1 xenograft mouse model reduced tumour size and enhanced survival. Critically, this was validated in patient-derived explants where enzymatic inactivation of IKBKE reduced cell proliferation and AR expression. Mechanistically, we provide evidence that IKBKE regulates AR levels via Hippo pathway inhibition to reduce c-MYC levels at cis-regulatory elements within the AR gene. Thus, IKBKE is a therapeutic target in advanced PC suggesting repurposing of clinically tested IKBKE inhibitors could be beneficial to castrate resistant PC patients.


Insulin-like growth factor-1 preconditioning accentuates intrinsic survival mechanism in stem cells to resist ischemic injury by orchestrating protein kinase cα-erk1/2 activation.
To test our hypothesis that the intrinsic molecular mechanism in stem cells for adaptation to ischemia is accentuated by preconditioning with insulin-like growth factor (IGF-1). Bone marrow Sca-1(+) cells were exposed to oxygen and glucose deprivation (OGD) for up to 12 h. Erk1/2 was activated in Sca-1(+) cells under OGD which was blocked by MEK inhibitor (PD98059) and resulted in accelerated cell death. Moreover, elevated intracellular calcium with concomitant activation of protein kinase C (PKC) was observed under OGD. Pretreatment with nifedipine or dantrolene reduced cellular calcium, abrogated PKC and Erk1/2 activation, and increased cytotoxicity. Treatment with phorbol 12-myristate 13-acetate (PMA) for 30 min (short-term) activated Erk1/2, whereas 12 h (long-term) PMA treatment abrogated PKCα, reduced Erk1/2 activation and significantly increased cell death under OGD. These results were confirmed by loss-of-function studies using PKCα and Erk1/2 specific small interfering RNA. Gain-of-function studies with PKCα plasmid transfection improved cell survival under OGD. Preconditioning with 100 nM IGF-1 accentuated the intrinsic mechanism of resistance of the cells to ischemia via Erk1/2 activation and improved their survival under OGD as well as post-transplantation in an experimentally infarcted heart. Strategies to target intrinsic survival mechanism in stem cells by growth factor preconditioning to enhance their survival via activation of PKCα and Erk1/2 are innovative. Intracellular calcium elevation under OGD activated PKCα and Erk1/2 as a part of the intrinsic prosurvival mechanism that was accentuated during preconditioning with IGF-1 to protect Sca-1(+) cells from ischemic injury.


Internet use, abuse, and dependence among students at a southeastern regional university.
To assess Internet use, abuse, and dependence. 411 undergraduate students. Ninety percent of participants reported daily Internet use. Approximately half of the sample met criteria for Internet abuse, and one-quarter met criteria for Internet dependence. Men and women did not differ on the mean amount of time accessing the Internet each day; however, the reasons for accessing the Internet differed between the 2 groups. Depression was correlated with more frequent use of the Internet to meet people, socially experiment, and participate in chat rooms, and with less frequent face-to-face socialization. In addition, individuals meeting criteria for Internet abuse and dependence endorsed more depressive symptoms, more time online, and less face-to-face socialization than did those not meeting the criteria. Mental health and student affairs professionals should be alert to the problems associated with Internet overuse, especially as computers become an integral part of college life.


Sedation during Minimal Invasive Surfactant Therapy in Preterm Infants.
There is no data available whether sedation should be given during minimally invasive surfactant therapy (MIST). To compare the level of comfort of preterm infants receiving sedation versus no sedation for MIST. A retrospective study of preterm infants receiving MIST was performed in Leiden University Medical Center in 2014. Sedation (propofol 1 mg/kg) was optional and left to the discretion of the caregiver. Standardized COMFORTneo scores were compared, and COMFORTneo <14 was considered comfortable. Basic characteristics and complications were noted. In 38 infants receiving MIST, 23 received propofol and 15 were not sedated. Mean (SD) gestational age [29 (2) vs. 29 (3) weeks] and birth weight [1,312 (483) vs. 1,469 (588) g] were not different. Median (IQR) COMFORTneo was not different between the groups before [11 (9-15) vs. 10 (8-12)] and after MIST [10 (8-12) vs. 9 (8-10)], but lower in the sedated group during MIST [12 (9-17) vs. 20 (15-23)] with more often COMFORTneo <14 (56 vs. 11%). Duration of MIST [2 (2-4) vs. 3 (2-7) min] and occurrence of bradycardia (13 vs. 33%) and hypotension (21 vs. 18%) were not different. Although not significant, intubation occurred more often in the sedated group (during MIST: 9 vs. 0%, <24 h after MIST: 26 vs. 13%). During MIST, oxygen saturation <80% lasted longer in the sedated group [3 (2-4) vs. 1 (0-2) min], and nasal intermittent positive pressure ventilation was applied more (100 vs. 33%). Preterm infants receiving MIST were more comfortable when sedation was given, but needed ventilation more often. A randomized controlled trial is warranted to test whether the benefit of sedation outweighs the risks of complications.


Generation of terahertz radiation from ionizing two-color laser pulses in Ar filled metallic hollow waveguides.
The generation of THz radiation from ionizing two-color femtosecond pulses propagating in metallic hollow waveguides filled with Ar is numerically studied. We observe a strong reshaping of the low-frequency part of the spectrum. More precisely, after several millimeters of propagation the spectrum is extended from hundreds of GHz up to approximately 150 THz. For longer propagation distances, nearly single-cycle near-infrared pulses with wavelengths around 4.5 microm are obtained by appropriate spectral filtering, with an efficiency of 0.1-1%.


High-luminosity blue and blue-green gallium nitride light-emitting diodes.
Compact and efficient sources of blue light for full color display applications and lighting eluded and tantalized researchers for many years. Semiconductor light sources are attractive owing to their reliability and amenability to mass manufacture. However, large band gaps are required to achieve blue color. A class of compound semiconductors formed by metal nitrides, GaN and its allied compounds AIGaN and InGaN, exhibits properties well suited for not only blue and blue-green emitters, but also for ultraviolet emitters and detectors. What thwarted engineers and scientists from fabricating useful devices from these materials in the past was the poor quality of material and lack of p-type doping. Both of these obstacles have recently been overcome to the point where highluminosity blue and blue-green light-emitting diodes are now available in the marketplace.


Mindboggling morphometry of human brains.
Mindboggle (http://mindboggle.info) is an open source brain morphometry platform that takes in preprocessed T1-weighted MRI data and outputs volume, surface, and tabular data containing label, feature, and shape information for further analysis. In this article, we document the software and demonstrate its use in studies of shape variation in healthy and diseased humans. The number of different shape measures and the size of the populations make this the largest and most detailed shape analysis of human brains ever conducted. Brain image morphometry shows great potential for providing much-needed biological markers for diagnosing, tracking, and predicting progression of mental health disorders. Very few software algorithms provide more than measures of volume and cortical thickness, while more subtle shape measures may provide more sensitive and specific biomarkers. Mindboggle computes a variety of (primarily surface-based) shapes: area, volume, thickness, curvature, depth, Laplace-Beltrami spectra, Zernike moments, etc. We evaluate Mindboggle's algorithms using the largest set of manually labeled, publicly available brain images in the world and compare them against state-of-the-art algorithms where they exist. All data, code, and results of these evaluations are publicly available.


Observer variability in mesothelioma tumor thickness measurements: defining minimally measurable lesions.
Single time-point unidimensional tumor thickness measurements define measurable disease for clinical trial inclusion and also constitute a field in the International Association for the Study of Lung Cancer prospective mesothelioma staging database. The modified Response Evaluation Criteria in Solid Tumors (RECIST) guidelines for mesothelioma did not alter the 10-mm minimum tumor measurement recommendation. However, as computed tomography technology has advanced, we sought to examine whether interobserver agreement was acceptable at smaller tumor thickness in mesothelioma. The primary observer selected 170 discrete measurement sites from 105 thoracic computed tomography scans from 50 consenting patients with pleural mesothelioma. Sites represented a range of tumor thickness, lesion morphology, and location. The outer (parietal) tumor margin was marked at each site and presented to five additional observers, who then selected the visceral margin of the tumor to create a line segment that captured tumor thickness. Relative differences among the observer measurements were estimated using a random-effects analysis of variance model to identify the smallest tumor thickness at which linear measurements could be made reliably. Systematic bias was observed, with some observers consistently measuring larger or smaller thicknesses than the thickness measured by others. The mean range across all 170 sites was 15.1% of the mean per-site measurement (SD = 9.1%; median range, 13.1%). Measurements acquired at sites with mean tumor thickness less than 7.5 mm demonstrated interobserver variability with a 75th percentile included 20% of the tumor thickness. The 95% confidence interval for relative interobserver measurement differences obtained for mean measurement lengths in the range 5 to 7.5 mm does not exceed the RECIST thresholds. This study has implications for the definition of minimally measurable tumor adopted by clinical trial and staging protocols. Although the statistical findings suggest that a reduction in "minimally measurable disease" from 10 mm to 5 or 7.5 mm might be warranted, clinical factors may ultimately dictate the most appropriate definition.


A History of United States Cannabis Law.
Perhaps the best way to understand early-Twenty-First Century state and federal cannabis law in the United States is to examine the relevant history. Justice Oliver Wendell Holmes, Jr.'s statement is apropos: "[A] page of history is worth a volume of logic." This article begins by discussing the early history of cannabis and its uses. Next, the article examines the first state and federal marijuana laws. After a brief comparison of alcohol prohibition to cannabis prohibition, this article addresses cannabis laws from the 1920s to the early 1950s. Then, the article takes up the reorganization of the federal drug regulatory bureaucracy since its inception. Addressing the current era of cannabis laws and regulations, this article recounts how marijuana became a Schedule I drug. The discussion then turns to changing social attitudes towards cannabis as reflected in presidential politics and popular culture. Starting with the late-1990s, this article describes the development of state and federal cannabis laws and policies up to the present day.


Building machines that learn and think like people.
Recent progress in artificial intelligence has renewed interest in building systems that learn and think like people. Many advances have come from using deep neural networks trained end-to-end in tasks such as object recognition, video games, and board games, achieving performance that equals or even beats that of humans in some respects. Despite their biological inspiration and performance achievements, these systems differ from human intelligence in crucial ways. We review progress in cognitive science suggesting that truly human-like learning and thinking machines will have to reach beyond current engineering trends in both what they learn and how they learn it. Specifically, we argue that these machines should (1) build causal models of the world that support explanation and understanding, rather than merely solving pattern recognition problems; (2) ground learning in intuitive theories of physics and psychology to support and enrich the knowledge that is learned; and (3) harness compositionality and learning-to-learn to rapidly acquire and generalize knowledge to new tasks and situations. We suggest concrete challenges and promising routes toward these goals that can combine the strengths of recent neural network advances with more structured cognitive models.


The plethysmograph: a review of recent literature.
There is much research to support the claim that the penile plethysmograph is a reliable and valid method of objectively measuring and assessing the erectile response in male sex offenders. The authors have reviewed some of the most recent literature dealing with this subject. The conclusion is that although the penile plethysmograph is the best objective measure of male sexual arousal and can be useful in the assessment and treatment of sex offenders, caution must be exercised because of the limitations. The limitations include using the plethysmograph as a predictive test, the lack of standards for administration and interpretation, the lack standardized norms, and the susceptibility to controlling results. Despite these limitations, the plethysmograph is often used as the single most important source of predictive data in some court rooms, boards of pardons, and for prison inmate classification systems. The plethysmograph is most effective in predictive situations when it is used in conjunction with multiple data source.


Having the rug pulled from under your feet: one project's experience of the US policy reversal on sex work.
After the election of President George W Bush in 2000, US government policy toward sexual and reproductive health changed dramatically. In May 2003, the Global AIDS Act was passed and prohibits allocation of US government funds to organizations that 'promote or advocate' legalization and practice of prostitution and sex trafficking. There are few documented examples of early impacts of this policy reversal on USAID-funded programmes already working with sex worker communities. This paper offers an anecdotal account of one programme in Cambodia that found itself caught in the ideological cross-fire of US politics, and describes consequent negative effects on the project's ability to offer appropriate and effective HIV prevention services to vulnerable migrant sex workers.


Outcome of intervention with or without low dose oral interferon alpha in thirty-two HIV-1 seropositive patients in a referral hospital.
Thirty two critically ill patients were admitted to Kenyatta National Hospital with a diagnosis of HIV-1 seropositivity on both ELISA and Western Blot between October 1989 and January 1990. Sixteen of the patients received low dose oral interferon alpha while the other 16 patients received intensive supportive management alone. Of the 16 patients who received low dose interferon alpha, 14 were discharged from the Hospital within 2 to 4 weeks of admission in a reasonable state of health while two of the patients died after 18 and 42 days from the time of admission. All the 16 patients who received intensive supportive management alone were dead within 4 weeks of entering the Hospital, their mean (+/- SD) stay after admission being 8.6 +/- 6.8 days. These results suggest that low dose oral interferon alpha may be of value in the care of critically ill HIV-1 seropositive patients.


Estimating Willingness-to-Pay for health insurance among rural poor in India by reference to Engel's law.
Community-Based Health Insurance (CBHI) (a.k.a. micro health insurance) is a contributory health insurance among rural poor in developing countries. As CBHI schemes typically function with no subsidy income, the schemes' expenditures cannot exceed their premium income. A good estimate of Willingness-To-Pay (WTP) among the target population affiliating on a voluntary basis is therefore essential for package design. Previous estimates of WTP reported materially and significantly different WTP levels across locations (even within one state), making it necessity to base estimates on household surveys. This is time-consuming and expensive. This study seeks to identify a coherent anchor for local estimation of WTP without having to rely on household surveys in each CBHI implementation. Using data collected in 2008-2010 among rural poor households in six locations in India (total 7874 households), we found that in all locations WTP expressed as percentage of income decreases with household income. This reminds of Engel's law on food expenditures. We checked several possible anchors: overall income, discretionary income and food expenditures. We compared WTP expressed as percentage of these anchors, by calculating the Coefficient of Variation (for inter-community variation) and Concentration indices (for intra-community variation). The Coefficient of variation was 0.36, 0.43 and 0.50 for WTP as percent of food expenditures, overall income and discretionary income, respectively. In all locations the concentration index for WTP as percentage of food expenditures was the lowest. Thus, food expenditures had the most consistent relationship with WTP within each location and across the six locations. These findings indicate that like food, health insurance is considered a necessity good even by people with very low income and no prior experience with health insurance. We conclude that the level of WTP could be estimated based on each community's food expenditures, and that this information can be obtained everywhere without having to conduct household surveys.


S-adenosyl methionine (SAMe) augmentation of serotonin reuptake inhibitors for antidepressant nonresponders with major depressive disorder: a double-blind, randomized clinical trial.
Despite the progressive increase in the number of antidepressants, many patients with major depressive disorder continue to be symptomatic. Clearly, there is an urgent need to develop better tolerated and more effective treatments for this disorder. The use of S-adenosyl methionine (SAMe), a naturally occurring molecule that serves as a methyl donor in human cellular metabolism, as adjunctive treatment for antidepressant nonresponders with major depressive disorder represents one such effort toward novel pharmacotherapy development. Participants were 73 serotonin reuptake inhibitor (SRI) nonresponders with major depressive disorder enrolled in a 6-week, double-blind, randomized trial of adjunctive oral SAMe (target dose: 800 mg/twice daily). Patients continued to receive their SRI treatment at a stable dose throughout the 6-week trial. The primary outcome measure for the study was the response rates according to the 17-item Hamilton Depression Rating Scale (HAM-D). The HAM-D response and remission rates were higher for patients treated with adjunctive SAMe (36.1% and 25.8%, respectively) than adjunctive placebo (17.6% versus 11.7%, respectively). The number needed to treat for response and remission was approximately one in six and one in seven, respectively. There was no statistically significant difference in the proportion of SAMe- versus placebo-treated patients who discontinued the trial for any reason (20.6% versus 29.5%, respectively), due to adverse events (5.1% versus 8.8%, respectively), or due to inefficacy (5.1% versus 11.7%, respectively). These preliminary results suggest that SAMe can be an effective, well-tolerated, and safe adjunctive treatment strategy for SRI nonresponders with major depressive disorder and warrant replication.


Development of new hepaticoenteric collateral pathways after hepatic arterial skeletonization in preparation for yttrium-90 radioembolization.
Development of new hepaticoenteric anastomotic vessels may occur after endovascular skeletonization of the hepatic artery. Left untreated, they can serve as pathways for nontarget radioembolization. The authors reviewed the incidence, anatomy, management, and significance of collateral vessel formation in patients undergoing radioembolization. One hundred thirty-eight treatments performed on 122 patients were reviewed. Each patient underwent a preparatory digital subtraction angiogram (DSA) and embolization of all hepaticoenteric vessels in preparation for yttrium-90 ((90)Y) administration. Successful skeletonization was verified by C-arm computed tomography (CACT) and technetium-99m macroaggregated albumin ((99m)TcMAA) scintigraphy. During the subsequent treatment session, DSA and CACT were repeated before administration of (90)Y, and the detection of extrahepatic perfusion prompted additional embolization. Forty-two patients (34.4%) undergoing 43 treatments (31.2%) required adjunctive embolization of hepaticoenteric vessels immediately before (90)Y administration. Previous scintigraphy findings showed extrahepatic perfusion in only three cases (7.1%). Vessels were identified by DSA in 54.1%, by CACT in 4.9%, or required both in 41.0%. The time interval between angiograms did not correlate with risk of requiring reembolization (P = .297). A total of 19.7% of vessels were new collateral vessels not visible during the initial angiography. Despite reembolization, three patients (7.1%) had gastric or duodenal ulceration, compared with 1.3% who never had visible collateral vessels, all of whom underwent whole-liver treatment with resin microspheres (P = .038). Development of collateral hepaticoenteric anastomoses occurs after endovascular skeletonization of the hepatic artery. Identified vessels may be managed by adjunctive embolization, but patients appear to remain at increased risk for gastrointestinal complications.


Earliest stone-tipped projectiles from the Ethiopian rift date to >279,000 years ago.
Projectile weapons (i.e. those delivered from a distance) enhanced prehistoric hunting efficiency by enabling higher impact delivery and hunting of a broader range of animals while reducing confrontations with dangerous prey species. Projectiles therefore provided a significant advantage over thrusting spears. Composite projectile technologies are considered indicative of complex behavior and pivotal to the successful spread of Homo sapiens. Direct evidence for such projectiles is thus far unknown from >80,000 years ago. Data from velocity-dependent microfracture features, diagnostic damage patterns, and artifact shape reported here indicate that pointed stone artifacts from Ethiopia were used as projectile weapons (in the form of hafted javelin tips) as early as >279,000 years ago. In combination with the existing archaeological, fossil and genetic evidence, these data isolate eastern Africa as a source of modern cultures and biology.


Multiresolution 3-D reconstruction from side-scan sonar images.
In this paper, a new method for the estimation of seabed elevation maps from side-scan sonar images is presented. The side-scan image formation process is represented by a Lambertian diffuse model, which is then inverted by a multiresolution optimization procedure inspired by expectation-maximization to account for the characteristics of the imaged seafloor region. On convergence of the model, approximations for seabed reflectivity, side-scan beam pattern, and seabed altitude are obtained. The performance of the system is evaluated against a real structure of known dimensions. Reconstruction results for images acquired by different sonar sensors are presented. Applications to augmented reality for the simulation of targets in sonar imagery are also discussed.


DNA polymerase beta: pre-steady-state kinetic analysis and roles of arginine-283 in catalysis and fidelity.
DNA polymerase beta (pol beta) is the smallest and least complex DNA polymerase. The structure of the enzyme is well understood, but little is known about its catalytic properties, particularly processivity and fidelity. Pre-steady-state analysis of the incorporation of a single nucleotide into a short 25/45 oligonucleotide primer-template by pol beta was used to define the kinetic parameters of the polymerase. In addition, nucleotide analogs and site-specific mutants, along with structural analyses, were used to probe the structure-function relationship of pol beta. Several significant findings have been obtained: (i) The catalysis by pol beta is processive and displays an initial burst under pre-steady-state conditions, but the processivity is poor compared to other polymerases. (ii) The fidelity of pol beta is also low relative to other polymerases. (iii) Under pre-steady-state conditions the chemical step appears to be only partially rate-limiting on the basis of the low thio effect (4.3), defined as kpol(dNTP)/kpol(dNTP alpha S). The thio effect increases to 9 for incorporation of an incorrect nucleotide. These results are consistent with the existence of a substrate-induced conformational change that is also partially rate-limiting. (iv) A comparison between the two-dimensional NMR spectra of the wild-type and mutant enzymes indicates that the mutations at position 283 did not significantly perturb the structure of the enzyme. The conformational stability of the mutants is also unperturbed. Thus, R283 is not important to the overall structure of the enzyme. (v) The results of kinetic analyses of R283A and R283K mutants indicate that the hydrogen bond between R283 of pol beta and the template is important for catalysis. Both R283A and R283K mutants displayed decreases in catalytic efficiency by a factor of ca. 200 relative to wild-type pol beta. The mutants are also less faithful by a factor of 2-4, in terms of the T-G mispair vs the T-A correct pair. The perturbation, however, could occur at both the implied conformational step and the chemical step, since the thio effects of the mutants for both correct and incorrect nucleotides are similar to those of WT pol beta.


Investigation of the Lith1 candidate genes ABCB11 and LXRA in human gallstone disease.
Genetic susceptibility in the causation of gallbladder diseases was recognized as early as 1937. A major gallstone susceptibility locus (Lith1) was identified in 1995 by quantitative trait locus mapping in mice. Two attractive positional and functional candidate genes in LXRA and ABCB11 are located in this interval. ABCB11 is associated with progressive familial cholestasis. This study was undertaken to investigate LXRA and ABCB11 as candidate genes for gallstone disease in humans. Eight hundred and ten patients who underwent cholecystectomy for symptomatic gallstone disease (median age of onset, 50 years) were compared with 718 sex-matched control individuals. Control individuals were sonographically free of gallstones. Haplotype tagging and all known coding single nucleotide polymorphisms (SNPs) were genotyped for ABCB11 (n=29) and LXRA (n=10). The investigated high-risk patient sample provides a power of greater than 80% for the detection of odds ratios down to 1.55. No evidence of association of the two genes in the single point tagging markers, coding variants or in the sliding window haplotype analysis was detected (all nominal single-point P values>or=.08). In conclusion, in the investigated German sample, no evidence of association of ABCB11 and LXRA to gallstone susceptibility was detected. The gallstone trait is not allelic to progressive familial cholestasis at the ABCB11 locus. Systematic fine mapping of the Lith1 region is required to identify the causative genetic variants for gallstone in mice and humans.


Prediction of biogeographical ancestry in admixed individuals.
Estimation of ancestral affiliation for human genotypes is now possible for major geographic populations and has been employed for forensic casework. Prediction algorithms, such as the Snipper Bayesian classifier, have the ability to classify non-admixed BGA in African (AFR), European (EUR), East Asian (EAS), and most Amerindian (NAM) individuals, but are not always appropriate for admixed individuals. Artificial admixture was simulated for all possible admixture ratios (1:1, 3:1, 2:1:1, and 1:1:1:1) from four grandparents. The simulated genotypes were used to test the accuracy of various prediction algorithms, most successful of which were the population genetics program, STRUCTURE, and a novel genetic distance algorithm (GDA). STRUCTURE was ideal for admixed individuals with 1:1 and 3:1 ratios from AFR, EUR, EAS, and NAM reference populations. Individuals with 1:1:1:1 BGA proportions were more accurately predicted by GDA. The use of hypothetical root genotypes improved the accuracy of GDA predictions for 1:1 and 3:1 admixtures and STRUCTURE classification of 1:1:1:1 admixture. The GDA requires only allele or genotype frequency values from each reference population, which offers a simpler sampling and input formatting procedure than is required by STRUCTURE. It can also be implemented in a spreadsheet without the need for long run times.


An examination of the Clinical Impairment Assessment among women at high risk for eating disorder onset.
Identifying measures that reliably and validly assess clinical impairment has important implications for eating disorder (ED) diagnosis and treatment. The current study examined the psychometric properties of the Clinical Impairment Assessment (CIA) in women at high risk for ED onset. Participants were 543 women (20.6 ± 2.0 years) who were classified into one of three ED categories: clinical ED, high risk for ED onset, and low risk control. Among high risk women, the CIA demonstrated high internal consistency (α = 0.93) and good convergent validity with disordered eating attitudes (rs = 0.27-0.68, ps < 0.001). Examination of the CIA's discriminant validity revealed that CIA global scores were highest among women with a clinical ED (17.7 ± 10.7) followed by high risk women (10.6 ± 8.5) and low risk controls (3.0 ± 3.3), respectively (p < 0.001). High risk women reporting behavioral indices of ED psychopathology (objective and/or subjective binge episodes, purging behaviors, driven exercise, and ED treatment history) had higher CIA global scores than those without such indices (ps < 0.05), suggesting good criterion validity. These data establish the first norms for the CIA in a United States sample. The CIA is psychometrically sound among high risk women, and heightened levels of impairment among these individuals as compared to low risk women verify the relevance of early intervention efforts.


Screening for cardiovascular disease risk using traditional risk factor assessment or coronary artery calcium scoring: the ROBINSCA trial.
Screening for a high cardiovascular disease (CVD) risk followed by preventive treatment can potentially reduce coronary heart disease-related morbidity and mortality. ROBINSCA (Risk Or Benefit IN Screening for CArdiovascular disease) is a population-based randomized controlled screening trial that investigates the effectiveness of CVD screening in asymptomatic participants using the Systematic COronary Risk Evaluation (SCORE) model or coronary artery calcium (CAC) scoring. This study describes the distributions in risk and treatment in the ROBINSCA trial. Individuals at expected elevated CVD risk were randomized into screening arm A (n = 14 478; SCORE, 10-year fatal and non-fatal risk); or screening arm B (n = 14 450; CAC scoring). Preventive treatment was largely advised according to current Dutch guidelines. Risk and treatment differences between the screening arms were analysed. A total of 12 185 participants (84.2%) in arm A and 12 950 (89.6%) in arm B were screened. In total, 48.7% were women, and median age was 62 (interquartile range 10) years. SCORE screening identified 45.1% at low risk (SCORE < 10%), 26.5% at intermediate risk (SCORE 10-20%), and 28.4% at high risk (SCORE ≥ 20%). According to CAC screening, 76.0% were at low risk (Agatston < 100), 15.1% at high risk (Agatston 100-399), and 8.9% at very high risk (Agatston ≥ 400). CAC scoring significantly reduced the number of individuals indicated for preventive treatment compared to SCORE (relative reduction women: 37.2%; men: 28.8%). We showed that compared to risk stratification based on SCORE, CAC scoring classified significantly fewer men and women at increased risk, and less preventive treatment was indicated. NTR6471.


HIV testing in primary care: feasibility and acceptability of provider initiated HIV testing and counseling for sub-Saharan African migrants.
Provider-initiated HIV testing and counseling (PITC) is recommended to reduce late HIV diagnoses, common among Sub-Saharan African migrants (SAM) residing in Europe. Primary care represents an ideal entry point for PITC. To support Flemish general practitioners (GPs), we developed a culturally sensitive PITC tool. Over a 12-week period, 65 GPs implemented PITC to assess acceptability and feasibility of PITC. The qualitative evaluation showed high acceptability among physicians. Routine PITC was challenged by physicians' personal discomfort, assumptions of patients' sexual risk, perceived incoherence with reasons for consultation, and time pressure. The best opportunity for PITC was an indicated blood analysis for other medical reasons. Counseling skills improved during the implementation, but participants still advocated for reduced counseling requirements. PITC proved to be feasible in primary care settings, but the up-scaling requires a reformulation of counseling guidelines, a policy stipulating the role of GPs in the prevention-care continuum, and an investment in (continuous) training.


Use of Doppler ultrasound for non-invasive urodynamic diagnosis.
A totally non-invasive transperineal urodynamic technique using Doppler ultrasonography has been developed. Since normal urine does not have blood cells, urine was thought not to produce the Doppler effects. However, basic studies confirmed that the decrease of pressure at high velocity (Bernouilli effect) caused dissolved gas to form microbubbles, which are detected by Doppler ultrasonography. Subjects sat and the probe was advanced via remote control to achieve gentle contact with the perineal skin. The digital uroflow data signals and the color Doppler ultrasound video images were processed on a personal computer. The flow-velocity curves from two sites; the distal prostatic urethra just above the external sphincter (V1) and the sphincteric urethra (V2) were plotted against time. The parameters of both the pressure-flow studies and the Doppler ultrasound urodynamic studies were compared in men who had various degrees of obstruction. Functional cross-sectional area at prostatic urethra (A1), calculated by Q(max)/V1, was lower in the group of bladder outlet obstruction (BOO) vs. control group. Velocity ratio (VR), which was calculated by V1/V2, was the parameter having the best correlation with BOO index, though A1 had a similar correlation. This method is viable to diagnose the degree of BOO. The development of non-invasive Doppler ultrasound videourodynamics (Doppler UDS) will dramatically expand the information on voiding function.


Proteomic analysis of temporally stimulated ovarian cancer cells for biomarker discovery.
While ovarian cancer remains the most lethal gynecological malignancy in the United States, there are no biomarkers available that are able to predict therapeutic responses to ovarian malignancies. One major hurdle in the identification of useful biomarkers has been the ability to obtain enough ovarian cancer cells from primary tissues diagnosed in the early stages of serous carcinomas, the most deadly subtype of ovarian tumor. In order to detect ovarian cancer in a state of hyperproliferation, we analyzed the implications of molecular signaling cascades in the ovarian cancer cell line OVCAR3 in a temporal manner, using a mass-spectrometry-based proteomics approach. OVCAR3 cells were treated with EGF(1), and the time course of cell progression was monitored based on Akt phosphorylation and growth dynamics. EGF-stimulated Akt phosphorylation was detected at 12 h post-treatment, but an effect on proliferation was not observed until 48 h post-exposure. Growth-stimulated cellular lysates were analyzed for protein profiles between treatment groups and across time points using iTRAQ labeling and mass spectrometry. The protein response to EGF treatment was identified via iTRAQ analysis in EGF-stimulated lysates relative to vehicle-treated specimens across the treatment time course. Validation studies were performed on one of the differentially regulated proteins, lysosomal-associated membrane protein 1 (LAMP-1), in human tissue lysates and ovarian tumor tissue sections. Further, tissue microarray analysis was performed to demarcate LAMP-1 expression across different stages of epithelial ovarian cancers. These data support the use of this approach for the efficient identification of tissue-based markers in tumor development related to specific signaling pathways. LAMP-1 is a promising biomarker for studies of the progression of EGF-stimulated ovarian cancers and might be useful in predicting treatment responses involving tyrosine kinase inhibitors or EGF receptor monoclonal antibodies.


Coupled cavity terahertz quantum cascade lasers with integrated emission monitoring.
We demonstrate the on-chip generation and detection of terahertz radiation in coupled cavity systems using a single semiconductor heterostructure. Multiple sections of a terahertz quantum cascade laser structure in a double-metal waveguide are optically coupled and operate either as a laser or an integrated emission monitor. A detailed analysis of the photon-assisted carrier transport in the active region below threshold reveals the detection mechanism for photons emitted by the very same structure above threshold. Configurations with a single laser cavity and two coupled laser cavities are studied. It is shown that the integrated detector can be used for spatial sensing of the light intensity within a coupled cavity.


Normal modes of the Indian elephant bell.
The geometrical structure of the Indian elephant bell is presented and the requirements on its normal modes from group representation theory are described. These are in good agreement with the results of a finite-element model (FEM) for a specific 16-tine case. The spectrum consists of a sequence of families of modes lying on saturation curves, completely different from those of conventional bells. Physical explanations for the occurrence of these families are presented in terms of the tines behaving as a closed loop of coupled cantilevers with constraints from the dome. Each family is found to consist of modes in one of two specific sequences of symmetry types. Experimental measurements of the modes of this same 16-tine bell, using electronic speckle pattern interferometry (ESPI), have been made and are compared with the FEM predictions. Although the interpretation of the interferograms is difficult in all but the simpler cases, agreement in terms of frequencies is surprisingly good for the first few family sequences. The ESPI study also showed up numerous harmonics and subharmonics of true normal modes, showing the system to be rather non-linear and making comparisons with the FEM results tricky.


Lactate dehydrogenase-elevating virus replication persists in liver, spleen, lymph node, and testis tissues and results in accumulation of viral RNA in germinal centers, concomitant with polyclonal activation of B cells.
Lactate dehydrogenase-elevating virus (LDV) replicates primarily and most likely solely in a subpopulation of macrophages in extraneuronal tissues. Infection of mice, regardless of age, with LDV leads to the rapid cytocidal replication of the virus in these cells, resulting in the release of large amounts of LDV into the circulation. The infection then progresses into life-long, asymptomatic, low-level viremic persistence, which is maintained by LDV replication in newly generated LDV-permissive cells which escapes all antiviral immune responses. In situ hybridization studies of tissue sections of adult FVB mice revealed that by 1 day postinfection (p.i.), LDV-infected cells were present in practically all tissues but were present in the highest numbers in the lymph nodes, spleen, and skin. In the central nervous system, LDV-infected cells were restricted to the leptomeninges. Most of the infected cells had disappeared at 3 days p.i., consistent with the cytocidal nature of the LDV infection, except for small numbers in lymph node, spleen, liver, and testis tissues. These tissues harbored infected cells until at least 90 days p.i. The results suggest that the generation of LDV-permissive cells during the persistent phase is restricted to these tissues. The continued presence of LDV-infected cells in testis tissue suggests the possibility of LDV release in semen and sexual transmission. Most striking was the accumulation of large amounts of LDV RNA in newly generated germinal centers of lymph nodes and the spleen. The LDV RNA was not associated with infected cells but was probably associated with virions or debris of infected, lysed cells. The appearance of LDV RNA in germinal centers in these mice coincided in time with the polyclonal activation of B cells, which leads to the accumulation of polyclonal immunoglobulin G2a and low-molecular-weight immune complexes in the circulation.


Neurochemical characterization of nerve fibers in the porcine gallbladder wall under physiological conditions and after the administration of Salmonella enteritidis lipopolysaccharides (LPS).
Lipopolysaccharides (LPS, bacterial endotoxin) are a component of the cellular membrane of Gram-negative bacteria, which is known as an important pathological factor. In spite of many previous studies describing multidirectional negative effects of LPS on living organisms, the knowledge concerning the influence of bacterial endotoxins on the gallbladder innervation is extremely scarce. The present study, based on the immunofluorescence technique, describes the changes in the neurochemical characterization of nerves within various parts of the porcine gallbladder (neck, body and fundus) after the administration of low doses of LPS. The obtained results show that even low doses of bacterial endotoxins affect the nerve structures within the gallbladder wall and the intensity of fluctuations in immunoreactivity to particular substances clearly depends on the part of the investigated organ. The most evident changes were observed in the case of fibers exhibiting the presence of neuropeptide Y (an increase from 7.84 ± 0.17 to 14.66 ± 0.37) in the neck, substance P (an increase from 0.88 ± 0.1 to 8.4 ± 0.3) in the body and the vesicular acetylocholine transporter in the gallbladder's fundus (an increase from 4.29 ± 0.18 to 11.01 ± 0.26). The mechanisms of the observed changes still remain unclear, but probably they are connected with the pro-inflammatory and/or neurodegenerative activity of LPS.


Isolation of Mycoplasma capricolum-like strains from chickens.
Members of the genus Mycoplasma infect a wide range of hosts, but individual Mycoplasma species tend to exhibit a considerable degree of host specificity. We characterized Mycoplasma strain 700, isolated from a kidney of a layer hen in Spain and Mycoplasma strains ULB-A and ULB-B, isolated from the air sac and from the bile of stunted broiler chickens in Slovenia. The serologic examination showed that these three strains are antigenically unrelated to all of the recognized Mycoplasma species of avian origin, but closely related to the ruminant mycoplasma Mycoplasma capricolum subspecies capricolum (M. capricolum). The comparison of their 16S rRNA gene sequences with the sequence of M. capricolum (California kid) revealed 99.66% sequence identity for the strain 700 and 99.59% identity for strains ULB-A and ULB-B. Moreover, the predicted DnaK sequences of the M. capricolum-like strains, isolated from chickens, were identical to DnaK sequences of M. capricolum. Comparison of their dnaK gene sequences with M. capricolum showed 99.64% sequence identity for strain 700 and 99.27% identity for strains ULB-A and ULB-B. In the flock from which M. capricolum-like strains ULB-A and ULB-B were isolated, the majority of chickens (83% of the chickens examined) raised antibodies reacting with M. capricolum antigens. Notably, the infection of chickens with M. capricolum-like strains represents an unusual exception to the range of Mycoplasma species host specificity.


Altered phosphorylation pattern of simian virus 40 T antigen expressed in insect cells by using a baculovirus vector.
The phosphorylation pattern of simian virus 40 (SV40) large tumor (T) antigen purified from insect cells infected with a recombinant baculovirus was compared with that reported previously for T antigen from SV40-infected monkey cells. The specific activity of metabolic phosphate labeling of baculovirus T antigen was reduced, and the phosphopeptide map of the baculovirus protein, while qualitatively similar to that of lytic T, revealed several quantitative differences. The most striking difference was the prominence in the baculovirus map of peptides containing phosphothreonine 124. These peptides are known to arise from other phosphopeptides upon dephosphorylation of neighboring serines, suggesting that baculovirus T may be underphosphorylated at these serines and perhaps other sites. Functional assays used to further investigate the phosphorylation state of the baculovirus protein included SV40 DNA binding after enzymatic dephosphorylation with alkaline phosphatase and after phosphorylation by a murine homolog of cdc2 protein kinase. The results imply that baculovirus T antigen is underphosphorylated, in particular at those serine residues whose phosphorylation is responsible for down regulation of DNA-binding activity at site II in the core origin of DNA replication. In contrast, no evidence for a functionally significant underphosphorylation at threonine 124 could be found.


Comparative analysis of clinical profile: Chronic cough vs paradoxical vocal fold motion.
Ongoing contention surrounding typical demographic and clinical attributes of chronic cough (CC) and paradoxical vocal fold motion (PVFM) impedes timely diagnosis and optimum patient care. Designed to reduce preventable patient morbidity through improved recognition and differentiation, the current study aimed to determine representative clinical profiles for CC and PVFM, with identification of distinctive attributes from the general population and risk factors associated with each diagnosis. Self-reported medical questionnaires, demographic and lifestyle attributes of CC and PVFM cases from a disease-specific outcomes database were compared to US population data and published normative values. Univariate comparison and multivariate regression modelling of age, sex, alcohol intake, smoking, Reflux Symptom Index (RSI), Voice Handicap Index (VHI), and Generalized Anxiety Disorder 7-item Scale (GAD-7) determined distinguishing features between the clinical groups, including odds ratios for presenting with CC versus PVFM. Clinical profiles developed from 283 (128 CC, 155 PVFM) adults (18-91 years) were significantly different from the general population across each demographic, lifestyle and clinical variable (all p < .01), with the exception of obesity. Age (55.39 ± 13.54 vs 45.07 ± 16.51 years, p < .01) and mean RSI score (21.5 ± 9.02 vs 18.1 ± 9.08, p < .01) most reliably distinguished CC from PVFM, with those aged 60-69 years (OR = 9.45) most likely to be diagnosed with CC. Standard clinical profiles of CC and PVFM are distinct from the general population, aiding determination of relative probabilities and risk factors in the differential diagnostic process. Variations between CC and PVFM were subtle, reliably distinguished by age and relative severity of laryngopharyngeal reflux symptomatology.


Accuracy of diagnostic biopsy for the histological subtype of malignant pleural mesothelioma.
Histological subtype is an established prognostic factor in malignant pleural mesothelioma (MPM). We retrospectively investigated the accuracy of classifying histological subtype on diagnostic biopsies and examined the impact of different diagnostic procedures on the outcome. Consecutive patients with histologically confirmed MPM who underwent extrapleural pneumonectomy (EPP) from 1994 to 2009 were included. Patient records were reviewed, and the initial diagnoses of histological subtype were obtained. The archival EPP specimens were reviewed by a panel of pathologists. The histological subtype obtained at review was compared with the initial diagnosis. Eighty-five patients underwent EPP. Two patients achieved a pathological complete response after neoadjuvant chemotherapy, leaving 83 patients to be included in this review. Different diagnostic methods were used before EPP: 81% thoracoscopy; 7% thoracotomy; 11% computed tomography-guided procedure; and 1% other. Patients determined to have an epithelial subtype (n = 64) at EPP were diagnosed correctly at initial diagnostic biopsy in 84% of cases, whereas patients considered to have a biphasic subtype (n = 19) at EPP were diagnosed correctly at diagnostic biopsy in 26% of cases. The sensitivity and specificity of diagnostic biopsy for epithelial MPM was 93% and 31%, respectively. The overall subtype misclassification rate was 20%. Biopsy by thoracotomy was most accurate in subtype classification (83%) compared with thoracoscopy (74%) and computed tomography-guided procedure (44%). The determination of histological subtype from a diagnostic biopsy is difficult due to sampling error, but an adequate specimen obtained from surgical biopsy increases the accuracy of subtype classification compared with radiological-guided biopsies.


Modulation of the face- and body-selective visual regions by the motion and emotion of point-light face and body stimuli.
Neural regions selective for facial or bodily form also respond to facial or bodily motion in highly form-degraded point-light displays. Yet it is unknown whether these face-selective and body-selective regions are sensitive to human motion regardless of stimulus type (faces and bodies) or to the specific motion-related cues characteristic of their proprietary stimulus categories. Using fMRI, we show that facial and bodily motions activate selectively those populations of neurons that code for the static structure of faces and bodies. Bodily (vs. facial) motion activated body-selective EBA bilaterally and right but not left FBA, irrespective of whether observers judged the emotion or color-change in point-light angry, happy and neutral stimuli. Facial (vs. bodily) motion activated face-selective right and left FFA, but only during emotion judgments for right FFA. Moreover, the strength of responses to point-light bodies vs. faces positively correlated with voxelwise selectivity for static bodies but not faces, whereas the strength of responses to point-light faces positively correlated with voxelwise selectivity for static faces but not bodies. Emotional content carried by point-light form-from-motion cues was sufficient to enhance the activity of several regions, including bilateral EBA and right FFA and FBA. However, although the strength of emotional modulation in right and left EBA by point-light body movements was related to the degree of voxelwise selectivity to static bodies but not static faces, there was no evidence that emotional modulation in fusiform cortex occurred in a similarly stimulus category-selective manner. This latter finding strongly constrains the claim that emotionally expressive movements modulate precisely those neuronal populations that code for the viewed stimulus category.


Clinical correlates of DHEA associated with post-traumatic stress disorder.
Increased plasma dehydroepiandrosterone (DHEA) and dehydroepiandrosterone-sulfate (DHEAS) have been demonstrated in post-traumatic stress disorder (PTSD), but the documented beneficial effects of these steroids in enhancing mood and cognition, as well as neuroprotection, suggest their presence in PTSD may be associated with defensive rather than maladaptive effects. We, therefore, examined plasma DHEA, DHEAS, cortisol, and the DHEA/cortisol ratio in 40 male veterans with or without PTSD, and determined their relationships to PTSD symptom severity and symptom improvement. The PTSD group showed significantly higher plasma DHEA and non-significantly higher DHEAS levels as well as a significantly lower cortisol/DHEA ratio, controlling for age. Regression analyses demonstrated that DHEA and DHEAS levels could be predicted by symptom improvement and coping, whereas the cortisol/DHEA ratio was predicted by severity of childhood trauma and current symptom severity. That greater symptom improvement was related to DHEA levels may suggest for a role for these hormones in modulating recovery from PTSD.


Optical manipulation of aerosol droplets using a holographic dual and single beam trap.
We present optical trapping and manipulation of pure water and salt water airborne droplets of various sizes ranging from sub-micrometers up to several tens of micrometers in a holographic dual and single beam trap. In the dual beam trap, successful fusion of droplets as well as precise delivery of many droplets and manipulation of multiple droplets are demonstrated. Furthermore, employing the transfer of the orbital angular momentum of light from Laguerre-Gaussian beams, we show that the water droplets orbit around the beam propagation axis and their tangential speed can be controlled by beam waist magnitude. We also demonstrate that sub-micrometer sized pure water droplets can be trapped and manipulated by a single beam trap with a relatively low numerical aperture. In this case, multiple stable trapping positions were observed, both theoretically and experimentally, which were due to the optical intensity oscillations in the focal region of the laser beam.


Engineered Resistance Against Papaya ringspot virus in Venezuelan Transgenic Papayas.
Local varieties of papaya grown in the Andean foothills of Mérida, Venezuela, were transformed independently with the coat protein (CP) gene from two different geographical Papaya ringspot virus (PRSV) isolates, designated VE and LA, via Agrobacterium tumefaciens. The CP genes of both PRSV isolates show 92 and 96% nucleotide and amino acid sequence similarity, respectively. Four PRSV-resistant R0 plants were intercrossed or selfed, and the progenies were tested for resistance against the homologous isolates VE and LA, and the heterologous isolates HA (Hawaii) and TH (Thailand) in greenhouse conditions. Resistance was affected by sequence similarity between the transgenes and the challenge viruses: resistance values were higher for plants challenged with the homologous isolates (92 to 100% similarity) than with the Hawaiian (94% similarity) and, lastly, Thailand isolates (88 to 89% similarity). Our results show that PRSV CP gene effectively protects local varieties of papaya against homologous and heterologous isolates of PRSV.


Cestodes from Hector's beaked whale ( Mesoplodon hectori ) and spectacled porpoise ( Phocoena dioptrica ) from Argentinean waters.
Single individuals of 2 little-known cetacean species, Mesoplodon hectori and Phocoena dioptrica , stranded and died on the coast of Argentina (Buenos Aires and Chubut provinces, respectively) and were studied for the presence of helminths. The cestodes found were described and illustrated using light microscopy. The following cestode taxa were recovered: Tetrabothrius ( Tetrabothrius ) hobergi n. sp. (several fragmented specimens, at least 1 gravid) and Tetrabothrius ( s.l. ) sp. 1 (several fragmented immature specimens) from M. hectori , and Tetrabothrius ( s.l. ) sp. 2 (single fragmented immature specimen) and 2 morphotypes of tetraphyllidean larvae from P. dioptrica. Tetrabothrius ( T. ) hobergi n. sp. can be distinguished from Tetrabothrius ( T. ) forsteri by the greater number of testes and larger eggs and oncospheres, from Tetrabothrius ( T. ) curilensis by the smaller testes and vitellarium, the shape and size of the ovary, and the larger oncospheres and longer embryonic hooks, and from Tetrabothrius ( T. ) sp. from Ziphius cavirostris by the narrower strobila, smaller scolex, and smaller number of testes. The generic designations of Tetrabothrius ( s.l. ) sp. 1 and Tetrabothrius ( s.l. ) sp. 2 were based on the scolex morphology. Tetrabothrius ( s.l. ) sp. 1 is closest to Tetrabothrius ( T. ) forsteri and Tetrabothrius ( Biamniculus ) innominatus based on the number of testes, while the scolex size of Tetrabothrius ( Tetrabothrius ) sp. 2 is within the variability range reported for Tetrabothrius ( T. ) forsteri . More definite identification of the 2 species was not possible due to the condition of the available material. The present study provides the first descriptions of cestodes from M. hectori and P. dioptrica , thus enriching the knowledge regarding the helminths of insufficiently studied marine mammals.


Effect of Sitagliptin on Kidney Function and Respective Cardiovascular Outcomes in Type 2 Diabetes: Outcomes From TECOS.
To evaluate chronic kidney disease (CKD) and cardiovascular outcomes in TECOS (Clinical trial reg. no. NCT00790205, clinicaltrials.gov) participants with type 2 diabetes and cardiovascular disease treated with sitagliptin, a dipeptidyl peptidase 4 inhibitor, according to baseline estimated glomerular filtration rate (eGFR). We used data from 14,671 TECOS participants assigned in a double-blind design to receive sitagliptin or placebo added to existing therapy, while aiming for glycemic equipoise between groups. Cardiovascular and CKD outcomes were evaluated over a median period of 3 years, with participants categorized at baseline into eGFR stages 1, 2, 3a, and 3b (≥90, 60-89, 45-59, or 30-44 mL/min/1.73 m2, respectively). Participants with eGFR stage 3b were older, were more often female, and had a longer duration of diabetes. Four-point major adverse cardiovascular event rates increased with lower baseline eGFR (3.52, 3.55, 5.74, and 7.34 events/100 patient-years for stages 1-3b, respectively). Corresponding adjusted hazard ratios for stages 2, 3a, and 3b versus stage 1 were 0.93 (95% CI 0.82-1.06), 1.28 (1.10-1.49), and 1.39 (1.13-1.72), respectively. Sitagliptin therapy was not associated with cardiovascular outcomes for any eGFR stage (interaction P values were all >0.44). Kidney function declined at the same rate in both treatment groups, with a marginally lower but constant eGFR difference (-1.3 mL/min/1.73 m2) in those participants who were assigned to sitagliptin. Treatment differences in these eGFR values remained after adjustment for region, baseline eGFR, baseline HbA1c, time of assessment, and within-study HbA1c levels. Impaired kidney function is associated with worse cardiovascular outcomes. Sitagliptin has no clinically significant impact on cardiovascular or CKD outcomes, irrespective of baseline eGFR.


Evaluation of a biodegradable synthetic hydrogel used as a guided bone regeneration membrane: an experimental study in dogs.
To test whether or not an experimental polyethylene glycol (PEG) membrane maintains the bone graft volume and contributes to the preservation of the ridge contour in comparison with a commercially available synthetic membrane. In 18 dogs, all mandibular premolars and the first molars were extracted. Ten weeks later, acute standardized defects were prepared. The defects of four dogs were randomly assigned to three modalities: (1) PEG plus deproteinized bovine bone mineral (DBBM) (PEG), (2) a resorbable glycolide trimethylene carbonate membrane plus DBBM (PGA-TMC), and (3) DBBM alone (DBBM). These dogs were then sacrificed for the baseline measurements. The remaining defects of 14 dogs were randomly assigned to (1) PEG plus DBBM, (2) PGA-TMC plus DBBM, (3) DBBM, and (4) empty defect. The dogs were sacrificed at baseline (n=4), 4 weeks (n=7), or at 16 weeks (n=7). Mixed model regressions and the non-parametric Brunner-Langer method were applied for statistical analysis. At baseline, equal tissue augmentation was observed in all groups. At 4 and 16 weeks, the greatest augmented area fractions were calculated for PEG (103%; 107%, respectively), followed by PGA-TMC (98%; 91%), DBBM (85%; 78%), and empty (46%; 54%), being statistically significant different (P<0.001) between PEG and empty at 4 and 16 weeks, and PEG and DBBM at 16 weeks. The overall decrease (P≤0.01) in the amount of bone graft between baseline and 16 weeks was -14% (PEG), -22% (PGA-TMC), and -23% (DBBM). The study demonstrates that the combination of the PEG membrane with DBBM maintains the bone graft volume over time better than controls. The PEG membrane with DBBM was also the most effective method to preserve the ridge contour.


The protein phosphatase 2A regulatory subunit PR70 is a gonosomal melanoma tumor suppressor gene.
Male gender is independently and significantly associated with poor prognosis in melanoma of all clinical stages. The biological underpinnings of this sex difference remain largely unknown, but we hypothesized that gene expression from gonosomes (sex chromosomes) might play an important role. We demonstrate that loss of the inactivated X chromosome in melanomas arising in females is strongly associated with poor distant metastasis-free survival, suggesting a dosage benefit from two X chromosomes. The gonosomal protein phosphatase 2 regulatory subunit B, beta (PPP2R3B) gene is located on the pseudoautosomal region (PAR) of the X chromosome in females and the Y chromosome in males. We observed that, despite its location on the PAR that predicts equal dosage across genders, PPP2R3B expression was lower in males than in females and was independently correlated with poor clinical outcome. PPP2R3B codes for the PR70 protein, a regulatory substrate-recognizing subunit of protein phosphatase 2A. PR70 decreased melanoma growth by negatively interfering with DNA replication and cell cycle progression through its role in stabilizing the cell division cycle 6 (CDC6)-chromatin licensing and DNA replication factor 1 (CDT1) interaction, which delays the firing of origins of DNA replication. Hence, PR70 functionally behaves as an X-linked tumor suppressor gene.


Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck.
We conducted a multinational, randomized study to compare radiotherapy alone with radiotherapy plus cetuximab, a monoclonal antibody against the epidermal growth factor receptor, in the treatment of locoregionally advanced squamous-cell carcinoma of the head and neck. Patients with locoregionally advanced head and neck cancer were randomly assigned to treatment with high-dose radiotherapy alone (213 patients) or high-dose radiotherapy plus weekly cetuximab (211 patients) at an initial dose of 400 mg per square meter of body-surface area, followed by 250 mg per square meter weekly for the duration of radiotherapy. The primary end point was the duration of control of locoregional disease; secondary end points were overall survival, progression-free survival, the response rate, and safety. The median duration of locoregional control was 24.4 months among patients treated with cetuximab plus radiotherapy and 14.9 months among those given radiotherapy alone (hazard ratio for locoregional progression or death, 0.68; P=0.005). With a median follow-up of 54.0 months, the median duration of overall survival was 49.0 months among patients treated with combined therapy and 29.3 months among those treated with radiotherapy alone (hazard ratio for death, 0.74; P=0.03). Radiotherapy plus cetuximab significantly prolonged progression-free survival (hazard ratio for disease progression or death, 0.70; P=0.006). With the exception of acneiform rash and infusion reactions, the incidence of grade 3 or greater toxic effects, including mucositis, did not differ significantly between the two groups. Treatment of locoregionally advanced head and neck cancer with concomitant high-dose radiotherapy plus cetuximab improves locoregional control and reduces mortality without increasing the common toxic effects associated with radiotherapy to the head and neck. (ClinicalTrials.gov number, NCT00004227.)


Effect Of A Large-Scale Social Franchising And Telemedicine Program On Childhood Diarrhea And Pneumonia Outcomes In India.
Despite the rapid growth of social franchising, there is little evidence on its population impact in the health sector. Similar in many ways to private-sector commercial franchising, social franchising can be found in sectors with a social objective, such as health care. This article evaluates the World Health Partners (WHP) Sky program, a large-scale social franchising and telemedicine program in Bihar, India. We studied appropriate treatment for childhood diarrhea and pneumonia and associated health care outcomes. We used multivariate difference-in-differences models to analyze data on 67,950 children ages five and under in 2011 and 2014. We found that the WHP-Sky program did not improve rates of appropriate treatment or disease prevalence. Both provider participation and service use among target populations were low. Our results do not imply that social franchising cannot succeed; instead, they underscore the importance of understanding factors that explain variation in the performance of social franchises. Our findings also highlight, for donors and governments in particular, the importance of conducting rigorous impact evaluations of new and potentially innovative health care delivery programs before investing in scaling them up.


In vitro neural differentiation of bone marrow stromal cells induced by cocultured olfactory ensheathing cells.
Bone marrow stromal cells (BMSCs) could be induced to differentiate into neural cells under certain conditions, nevertheless, optimal protocols that could be reproducible and reliable in generating transplantable BMSCs in vitro are still not available. We studied for the first time the neural differentiation of BMSCs induced by coculturing with olfactory ensheathing cells (OECs). BMSCs and OECs were isolated from bone marrow and nasal olfactory lamina propria of adult SD rats respectively, then brought to coculture with transwell culture dishes. At various time points (0h, 6h, 12h, 24h, 72h, 1 week and 2 weeks post-coculture), BMSCs were morphologically observed and processed for immunofluorescence and reverse transcription-polymerase chain reaction (RT-PCR). The number of cells assuming neural morphology dramatically increased at 1- and 2-week-post-coculture, so as the number of immunoreactive cells labeled by neural markers NSE, beta-III-tubulin, MAP2, GFAP and p75(NTR). Our findings demonstrate that BMSCs can efficiently differentiate into neural cells when coculturing with OECs, and the present protocol provides an alternative neurogenesis pathway for generating sufficient numbers of neural cells from BMSCs.


Infectious morbidity after radical vulvectomy.
This retrospective investigation describes the infectious morbidity of patients following radical vulvectomy with or without inguinal lymph node dissection. The charts of patients undergoing radical vulvectomy between January 1, 1986, and September 1, 1989, were reviewed for age, weight, cancer type, tumor stage, operative procedure(s), prophylactic antibiotic and its length of use, febrile morbidity, infection site, culture results, significant medical history, and length of use and number of drains or catheters used. The study group was composed of 61 patients, 14 of whom underwent a radical vulvectomy and 47 who also had inguinal lymph node dissection performed. Twenty-nine patients (48%) had at least 1 postoperative infection. Five patients (8%) had 2 or more postoperative infections. The site and incidence of the infections were as follows: urinary tract 23%, wound 23%, lymphocyst 3%, lymphatics (lymphangitis) 5%, and bowel (pseudomembranous colitis) 3%. The most common pathogens isolated from both urine and wound sites were Pseudomonas aeruginosa, enterococcus, and Escherichia coli. A significant decrease in wound infection was demonstrated when separate incisions were made for inguinal lymph node dissection (P <0.05). The mean number of days to onset of postoperative infection for wound, urine, lymphatics, lymphocyst, and bowel were 11, 8, 57, 48, and 5, respectively. We conclude that the clinical appearance of post-radical vulvectomy infections is delayed when compared with other post-surgical wound infections. Second, utilizing separate inguinal surgical incisions may reduce infectious morbidity. Finally, tumor stage and type do not necessarily increase the infectious morbidity of radical vulvar surgery.


Expansion of dysfunctional Tim-3-expressing effector memory CD8+ T cells during simian immunodeficiency virus infection in rhesus macaques.
The T cell Ig- and mucin domain-containing molecule-3 (Tim-3) negative immune checkpoint receptor demarcates functionally exhausted CD8(+) T cells arising from chronic stimulation in viral infections like HIV. Tim-3 blockade leads to improved antiviral CD8(+) T cell responses in vitro and, therefore, represents a novel intervention strategy to restore T cell function in vivo and protect from disease progression. However, the Tim-3 pathway in the physiologically relevant rhesus macaque SIV model of AIDS remains uncharacterized. We report that Tim-3(+)CD8(+) T cell frequencies are significantly increased in lymph nodes, but not in peripheral blood, in SIV-infected animals. Tim-3(+)PD-1(+)CD8(+) T cells are similarly increased during SIV infection and positively correlate with SIV plasma viremia. Tim-3 expression was found primarily on effector memory CD8(+) T cells in all tissues examined. Tim-3(+)CD8(+) T cells have lower Ki-67 content and minimal cytokine responses to SIV compared with Tim-3(-)CD8(+) T cells. During acute-phase SIV replication, Tim-3 expression peaked on SIV-specific CD8(+) T cells by 2 wk postinfection and then rapidly diminished, irrespective of mutational escape of cognate Ag, suggesting non-TCR-driven mechanisms for Tim-3 expression. Thus, rhesus Tim-3 in SIV infection partially mimics human Tim-3 in HIV infection and may serve as a novel model for targeted studies focused on rejuvenating HIV-specific CD8(+) T cell responses.


Goal conflict, goal facilitation, and health professionals' provision of physical activity advice in primary care: an exploratory prospective study.
The theory of planned behaviour has well-evidenced utility in predicting health professional behaviour, but focuses on a single behaviour isolated from the numerous potentially conflicting and facilitating goal-directed behaviours performed alongside. Goal conflict and goal facilitation may influence whether health professionals engage in guideline-recommended behaviours, and may supplement the predictive power of the theory of planned behaviour. We hypothesised that goal facilitation and goal conflict contribute to predicting primary care health professionals' provision of physical activity advice to patients with hypertension, over and above predictors of behaviour from the theory of planned behaviour. Using a prospective predictive design, at baseline we invited a random sample of 606 primary care health professionals from all primary care practices in NHS Grampian and NHS Tayside (Scotland) to complete postal questionnaires. Goal facilitation and goal conflict were measured alongside theory of planned behaviour constructs at baseline. At follow-up six months later, participants self-reported the number of patients, out of those seen in the preceding two weeks, to whom they provided physical activity advice. Forty-four primary care physicians and nurses completed measures at both time points (7.3% response rate). Goal facilitation and goal conflict improved the prediction of behaviour, accounting for substantial additional variance (5.8% and 8.4%, respectively) in behaviour over and above intention and perceived behavioural control. Health professionals' provision of physical activity advice in primary care can be predicted by perceptions about how their conflicting and facilitating goal-directed behaviours help and hinder giving advice, over and above theory of planned behaviour constructs. Incorporating features of multiple goal pursuit into the theory of planned behaviour may help to better understand health professional behaviour.


Compact modulated and tunable microdisk laser using vertical coupling and a feedback loop.
A study of vertical coupling conditions between microdisk based resonators and waveguides is presented using an analytical model. The coupling efficiency optimization is investigated and compared to 3D FDTD computations. We also demonstrate that coupling losses can be exploited to favor high quality factor modes in circular resonators. In addition, we propose to modify the shape of the coupled waveguide to enhance mode selectivity and obtain a very compact structure with mode hoping capacities. Lower thresholds and modulation are also expected.


A Validated HPTLC-Densitometric Method for Assay of Aucubin in Vitexagnus-castusL.
Vitexagnus-castusL. is a medicinal plant which is used in several dosage forms for women hormonal disorders and standardized according to the iridoids or flavonoids content. Aucubin, an iridoid glycoside, considered as a marker in some formulations. In this research, a thin layer chromatographic method with densitometric detection has been developed for quantitative determination of aucubin in chaste tree fruits. Chromatographic separation was performed using silica gel high performance thin-layer chromatography (HPTLC) plates with ethyl acetate-methanol-water 77 : 15 : 8 as mobile phase. Chromatograms were visualized using p-dimethylaminobenzaldehyde as reagent. Aucubin RF-value was about 0.5 and spots were scanned at 580 nm through a mercury lamp. By using this method, the amount of aucubin was found 43.5 mg/100 g of dried plant fruits. The method was validated for selectivity, linearity (r(2) = 0.997, 20-100 μg/mL), precision (intra-day < 4.9, inter-day < 7.2) and accuracy measured via determination of recovery (95-98%). The limit of detection and limit of quantization were found 6.6 and 20 μg/mL, respectively. This methodology was found to be precise with respect to the validation parameters. It is simple and convenient and could be applicable to the routine determination of aucubin in different Vitexagnus-castusL. samples.


progressiveMauve: multiple genome alignment with gene gain, loss and rearrangement.
Multiple genome alignment remains a challenging problem. Effects of recombination including rearrangement, segmental duplication, gain, and loss can create a mosaic pattern of homology even among closely related organisms. We describe a new method to align two or more genomes that have undergone rearrangements due to recombination and substantial amounts of segmental gain and loss (flux). We demonstrate that the new method can accurately align regions conserved in some, but not all, of the genomes, an important case not handled by our previous work. The method uses a novel alignment objective score called a sum-of-pairs breakpoint score, which facilitates accurate detection of rearrangement breakpoints when genomes have unequal gene content. We also apply a probabilistic alignment filtering method to remove erroneous alignments of unrelated sequences, which are commonly observed in other genome alignment methods. We describe new metrics for quantifying genome alignment accuracy which measure the quality of rearrangement breakpoint predictions and indel predictions. The new genome alignment algorithm demonstrates high accuracy in situations where genomes have undergone biologically feasible amounts of genome rearrangement, segmental gain and loss. We apply the new algorithm to a set of 23 genomes from the genera Escherichia, Shigella, and Salmonella. Analysis of whole-genome multiple alignments allows us to extend the previously defined concepts of core- and pan-genomes to include not only annotated genes, but also non-coding regions with potential regulatory roles. The 23 enterobacteria have an estimated core-genome of 2.46Mbp conserved among all taxa and a pan-genome of 15.2Mbp. We document substantial population-level variability among these organisms driven by segmental gain and loss. Interestingly, much variability lies in intergenic regions, suggesting that the Enterobacteriacae may exhibit regulatory divergence. The multiple genome alignments generated by our software provide a platform for comparative genomic and population genomic studies. Free, open-source software implementing the described genome alignment approach is available from http://gel.ahabs.wisc.edu/mauve.


Suppression of skeletal muscle lipoprotein lipase activity during physical inactivity: a molecular reason to maintain daily low-intensity activity.
We have examined the regulation of lipoprotein lipase (LPL) activity in skeletal muscle during physical inactivity in comparison to low-intensity contractile activity of ambulatory controls. From studies acutely preventing ambulatory activity of one or both the hindlimbs in rats, it was shown that approximately 90-95 % of the heparin-releasable (HR) LPL activity normally present in rat muscle with ambulatory activity is lost, and thus dependent on local contractile activity. Similarly, approximately 95 % of the differences in LPL activity between muscles of different fibre types was dependent on ambulatory activity. The robustness of the finding that physical inactivity significantly decreases muscle LPL activity was evident from confirmatory studies with different models of inactivity, in many rats and mice, both sexes, three muscle types and during both acute and chronic (11 days) treatment. Inactivity caused a local reduction of plasma [3H]triglyceride uptake into muscle and a decrease in high density lipoprotein cholesterol concentration. LPL mRNA was not differentially expressed between ambulatory controls and either the acutely or chronically inactive groups. Instead, the process involved a rapid loss of the HR-LPL protein mass (the portion of LPL largely associated with the vascular endothelium) by an actinomycin D-sensitive signalling mechanism (i.e. transcriptionally dependent process). Significant decreases of intracellular LPL protein content lagged behind the loss of HR-LPL protein. Treadmill walking raised LPL activity approximately 8-fold (P < 0.01) within 4 h after inactivity. The striking sensitivity of muscle LPL to inactivity and low-intensity contractile activity may provide one piece of the puzzle for why inactivity is a risk factor for metabolic diseases and why even non-vigorous activity provides marked protection against disorders involving poor lipid metabolism.


Recent advances in understanding the role of diet and obesity in the development of colorectal cancer.
Colorectal cancer (CRC) is a major cause of premature death in the UK and many developed countries. However, the risk of developing CRC is well recognised to be associated not only with diet but also with obesity and lack of exercise. While epidemiological evidence shows an association with factors such as high red meat intake and low intake of vegetables, fibre and fish, the mechanisms underlying these effects are only now being elucidated. CRC develops over many years and is typically characterised by an accumulation of mutations, which may arise as a consequence of inherited polymorphisms in key genes, but more commonly as a result of spontaneously arising mutations affecting genes controlling cell proliferation, differentiation, apoptosis and DNA repair. Epigenetic changes are observed throughout the progress from normal morphology through formation of adenoma, and the subsequent development of carcinoma. The reasons why this accumulation of loss of homoeostatic controls arises are unclear but chronic inflammation has been proposed to play a central role. Obesity is associated with increased plasma levels of chemokines and adipokines characteristic of chronic systemic inflammation, and dietary factors such as fish oils and phytochemicals have been shown to have anti-inflammatory properties as well as modulating established risk factors such as apoptosis and cell proliferation. There is also some evidence that diet can modify epigenetic changes. This paper briefly reviews the current state of knowledge in relation to CRC development and considers evidence for potential mechanisms by which diet may modify risk.


A 26 kd calcium binding protein from bovine rod outer segments as modulator of photoreceptor guanylate cyclase.
The resynthesis of cGMP in vertebrate photoreceptors by guanylate cyclase is one of the key events leading to the reopening of cGMP-gated channels after photoexcitation. Guanylate cyclase activity in vertebrate rod outer segments is dependent on the free calcium concentration. The basal activity of the enzyme observed at high concentrations of free calcium (greater than 0.5 microM) increases when the free calcium concentration is lowered into the nanomolar range (less than 0.1 microM). This effect of calcium is known to be mediated by a soluble calcium-sensitive protein in a highly cooperative way. We here show that this soluble protein, i.e. the modulator of photoreceptor guanylate cyclase, is a 26 kd protein. Reconstitution of the purified 26 kd protein with washed rod outer segment membranes containing guanylate cyclase revealed a 3- to 4-fold increase of cyclase activity when the free calcium concentration was lowered in the physiological range from 0.5 microM to 4 nM. Guanylate cyclase in whole rod outer segments was stimulated 10-fold in the same calcium range. The activation process in the reconstituted system was similar to the one in the native rod outer segment preparation, it showed a high cooperativity with a Hill coefficient n between 1.4 and 3.5. The half-maximal activation occurred between 110 and 220 nM free calcium. The molar ratio of the modulator to rhodopsin is 1:76 +/- 32. The protein is a calcium binding protein as detected with 45Ca autoradiography. Partial amino acid sequence analysis revealed a 60% homology to visinin from chicken cones.


Pore water colloid properties in argillaceous sedimentary rocks.
The focus of this work is to evaluate the colloid nature, concentration and size distribution in the pore water of Opalinus Clay and other sedimentary host rocks identified for a potential radioactive waste repository in Switzerland. Because colloids could not be measured in representative undisturbed porewater of these host rocks, predictive modelling based on data from field and laboratory studies is applied. This approach allowed estimating the nature, concentration and size distributions of the colloids in the pore water of these host rocks. As a result of field campaigns, groundwater colloid concentrations are investigated on the basis of their size distribution quantified experimentally using single particle counting techniques. The colloid properties are estimated considering data gained from analogue hydrogeochemical systems ranging from mylonite features in crystalline fissures to sedimentary formations. The colloid concentrations were analysed as a function of the alkaline and alkaline earth element concentrations. Laboratory batch results on clay colloid generation from compacted pellets in quasi-stagnant water are also reported. Experiments with colloids in batch containers indicate that the size distribution of a colloidal suspension evolves toward a common particle size distribution independently of initial conditions. The final suspension size distribution was found to be a function of the attachment factor of the colloids. Finally, calculations were performed using a novel colloid distribution model based on colloid generation, aggregation and sedimentation rates to predict under in-situ conditions what makes colloid concentrations and size distributions batch- or fracture-size dependent. The data presented so far are compared with the field and laboratory data. The colloid occurrence, stability and mobility have been evaluated for the water of the considered potential host rocks. In the pore water of the considered sedimentary host rocks, the clay colloid concentration is expected to be very low (<1ppb, for 10-100nm) which restricts their relevance for radionuclide transport.


FM-test: a fuzzy-set-theory-based approach to differential gene expression data analysis.
Microarray techniques have revolutionized genomic research by making it possible to monitor the expression of thousands of genes in parallel. As the amount of microarray data being produced is increasing at an exponential rate, there is a great demand for efficient and effective expression data analysis tools. Comparison of gene expression profiles of patients against those of normal counterpart people will enhance our understanding of a disease and identify leads for therapeutic intervention. In this paper, we propose an innovative approach, fuzzy membership test (FM-test), based on fuzzy set theory to identify disease associated genes from microarray gene expression profiles. A new concept of FM d-value is defined to quantify the divergence of two sets of values. We further analyze the asymptotic property of FM-test, and then establish the relationship between FM d-value and p-value. We applied FM-test to a diabetes expression dataset and a lung cancer expression dataset, respectively. Within the 10 significant genes identified in diabetes dataset, six of them have been confirmed to be associated with diabetes in the literature and one has been suggested by other researchers. Within the 10 significantly overexpressed genes identified in lung cancer data, most (eight) of them have been confirmed by the literatures which are related to the lung cancer. Our experiments on synthetic datasets show that FM-test is effective and robust. The results in diabetes and lung cancer datasets validated the effectiveness of FM-test. FM-test is implemented as a Web-based application and is available for free at http://database.cs.wayne.edu/bioinformatics.


Preparation and evaluation of liquid-crystal formulations with skin-permeation-enhancing abilities for entrapped drugs.
The usefulness of liquid crystals (LC) in topical formulations for application to skin was evaluated by measuring the in vitro permeation profile of a model compound, calcein, entrapped in a LC formulation, through excised hairless rat skin and a three-dimensional cultured human-skin model; the viability was determined using the MTT assay. Two physically stable LCs were prepared from a mixture of mono-, di-, and tri-esters 1, and monoesters 2, composed of erythritol and phytanylacetic acid. Cryo-transmission electron microscopy (cryo-TEM), electron diffraction patterns, and small-angle X-ray diffraction (SAXS) observations of the LC nanodispersions showed that the structures of the LCs were reverse hexagonal (LC-A) and cubic (LC-B). The skin-permeation properties of calcein were enhanced by entrapping in the LCs as a result of the increase in calcein partition from the LC dispersion solution into the skin; the properties were analyzed using a skin-permeation-time profile. Drug partitioning could also be modified by the LC structure. No skin damage was caused by the LC formulation in these experiments.The present study suggests that LC dispersions are potential additives in topical drug formulations and cosmetic formulations.


Evidence for invasion of a human oral cell line by Actinobacillus actinomycetemcomitans.
Actinobacillus actinomycetemcomitans, an oral bacterial species associated with periodontal disease, was found to invade human cell lines. Invasion was demonstrated by recovery of viable organisms from gentamicin-treated KB cell monolayers and by light and electron microscopy. Internalization occurred through a cytochalasin D-sensitive process. Invasion efficiencies of some A. actinomycetemcomitans strains were comparable to those of invasive members of the family Enterobacteriaceae. Differences in invasiveness were correlated with bacterial colonial morphology. Smooth variants invaded more proficiently than rough variants. A. actinomycetemcomitans can undergo a smooth-to-rough colonial morphology shift which results in the loss of invasiveness. Coordinated regulation of genes involved in the rough-to-smooth phenotypic transitions may play a role in the episodic nature of periodontal disease.


Single Upconversion Nanoparticle-Bacterium Cotrapping for Single-Bacterium Labeling and Analysis.
Detecting and analyzing pathogenic bacteria in an effective and reliable manner is crucial for the diagnosis of acute bacterial infection and initial antibiotic therapy. However, the precise labeling and analysis of bacteria at the single-bacterium level are a technical challenge but very important to reveal important details about the heterogeneity of cells and responds to environment. This study demonstrates an optical strategy for single-bacterium labeling and analysis by the cotrapping of single upconversion nanoparticles (UCNPs) and bacteria together. A single UCNP with an average size of ≈120 nm is first optically trapped. Both ends of a single bacterium are then trapped and labeled with single UCNPs emitting green light. The labeled bacterium can be flexibly moved to designated locations for further analysis. Signals from bacteria of different sizes are detected in real time for single-bacterium analysis. This cotrapping method provides a new approach for single-pathogenic-bacterium labeling, detection, and real-time analysis at the single-particle and single-bacterium level.


Near-patient testing of potassium levels using arterial blood gas analysers: can we trust these results?
Near-patient testing allows rapid availability of results to enable prompt decision-making. Potassium abnormalities are common in acutely ill patients and can be associated with life-threatening complications. At times there is uncertainty whether clinical decisions can be based on the potassium result obtained from arterial blood gas (ABG) analysers or if laboratory values should be awaited. To determine the opinion of clinicians regarding the use of blood gas analysers to measure potassium and to determine the level of agreement between blood gas analyser and laboratory measurements of potassium in arterial blood samples. Survey of 64 doctors using a questionnaire and a retrospective comparative study of 529 paired results of ABG and arterial laboratory measurements of potassium in 121 critically ill patients. 51.6% of the doctors would wait for laboratory confirmation and 48.4% would base clinical decisions on results obtained from the blood gas analyser. The difference between the means of potassium values from the two methods is 0.03 mmol/l (95% CI 0.011 to 0.056; p = 0.0041). The 95% limits of agreement were from -0.485 mmol/l (95% CI -0.524 to -0.447) to 0.551 mmol/l (95% CI 0.513 to 0.590). 95% of the results fell within the difference limits of 0.5 mmol/l. Most clinicians still await laboratory confirmation of results obtained from blood gas analysers but in this setting there is sufficient agreement between the results obtained from the authors' blood gas analyser and a laboratory analyser to enable effective clinical decisions to be made.


Computational identification and functional validation of regulatory motifs in cartilage-expressed genes.
Chondrocyte gene regulation is important for the generation and maintenance of cartilage tissues. Several regulatory factors have been identified that play a role in chondrogenesis, including the positive transacting factors of the SOX family such as SOX9, SOX5, and SOX6, as well as negative transacting factors such as C/EBP and delta EF1. However, a complete understanding of the intricate regulatory network that governs the tissue-specific expression of cartilage genes is not yet available. We have taken a computational approach to identify cis-regulatory, transcription factor (TF) binding motifs in a set of cartilage characteristic genes to better define the transcriptional regulatory networks that regulate chondrogenesis. Our computational methods have identified several TFs, whose binding profiles are available in the TRANSFAC database, as important to chondrogenesis. In addition, a cartilage-specific SOX-binding profile was constructed and used to identify both known, and novel, functional paired SOX-binding motifs in chondrocyte genes. Using DNA pattern-recognition algorithms, we have also identified cis-regulatory elements for unknown TFs. We have validated our computational predictions through mutational analyses in cell transfection experiments. One novel regulatory motif, N1, found at high frequency in the COL2A1 promoter, was found to bind to chondrocyte nuclear proteins. Mutational analyses suggest that this motif binds a repressive factor that regulates basal levels of the COL2A1 promoter.


Evidence-based clinical practice guideline on nonrestorative treatments for carious lesions: A report from the American Dental Association.
An expert panel convened by the American Dental Association Council on Scientific Affairs and the Center for Evidence-Based Dentistry conducted a systematic review and formulated evidence-based clinical recommendations for the arrest or reversal of noncavitated and cavitated dental caries using nonrestorative treatments in children and adults. The authors conducted a systematic search of the literature in MEDLINE and Embase via Ovid, Cochrane CENTRAL, and Cochrane database of systematic reviews to identify randomized controlled trials reporting on nonrestorative treatments for noncavitated and cavitated carious lesions. The authors used the Grading of Recommendations Assessment, Development and Evaluation approach to assess the certainty in the evidence and move from the evidence to the decisions. The expert panel formulated 11 clinical recommendations, each specific to lesion type, tooth surface, and dentition. Of the most effective interventions, the panel provided recommendations for the use of 38% silver diamine fluoride, sealants, 5% sodium fluoride varnish, 1.23% acidulated phosphate fluoride gel, and 5,000 parts per million fluoride (1.1% sodium fluoride) toothpaste or gel, among others. The panel also provided a recommendation against the use of 10% casein phosphopeptide-amorphous calcium phosphate. Although the recommended interventions are often used for caries prevention, or in conjunction with restorative treatment options, these approaches have shown to be effective in arresting or reversing carious lesions. Clinicians are encouraged to prioritize use of these interventions based on effectiveness, safety, and feasibility.


Asymmetric Processing of Numerical and Nonnumerical Magnitudes in the Brain: An fMRI Study.
It is well established that, when comparing nonsymbolic magnitudes (e.g., dot arrays), adults can use both numerical (i.e., the number of items) and nonnumerical (density, total surface areas, etc.) magnitudes. It is less clear which of these magnitudes is more salient or processed more automatically. In this fMRI study, we used a nonsymbolic comparison task to ask if different brain areas are responsible for the automatic processing of numerical and nonnumerical magnitudes, when participants were instructed to attend to either the numerical or the nonnumerical magnitudes of the same stimuli. An interaction of task (numerical vs. nonnumerical) and congruity (congruent vs. incongruent) was found in the right TPJ. Specifically, this brain region was more strongly activated during numerical processing when the nonnumerical magnitudes were negatively correlated with numerosity (incongruent trials). In contrast, such an interference effect was not evident during nonnumerical processing when the task-irrelevant numerical magnitude was incongruent. In view of the role of the right TPJ in the control of stimulus-driven attention, we argue that these data demonstrate that the processing of nonnumerical magnitudes is more automatic than that of numerical magnitudes and that, therefore, the influence of numerical and nonnumerical variables on each other is asymmetrical.