Organic residue analysis shows sub-regional patterns in the use of pottery by Northern European hunter-gatherers.
The introduction of pottery vessels to Europe has long been seen as closely linked with the spread of agriculture and pastoralism from the Near East. The adoption of pottery technology by hunter-gatherers in Northern and Eastern Europe does not fit this paradigm, and its role within these communities is so far unresolved. To investigate the motivations for hunter-gatherer pottery use, here, we present the systematic analysis of the contents of 528 early vessels from the Baltic Sea region, mostly dating to the late 6th-5th millennium cal BC, using molecular and isotopic characterization techniques. The results demonstrate clear sub-regional trends in the use of ceramics by hunter-gatherers; aquatic resources in the Eastern Baltic, non-ruminant animal fats in the Southeastern Baltic, and a more variable use, including ruminant animal products, in the Western Baltic, potentially including dairy. We found surprisingly little evidence for the use of ceramics for non-culinary activities, such as the production of resins. We attribute the emergence of these sub-regional cuisines to the diffusion of new culinary ideas afforded by the adoption of pottery, e.g. cooking and combining foods, but culturally contextualized and influenced by traditional practices.


Predictors of septic metastatic infection and mortality among patients with Klebsiella pneumoniae liver abscess.
Primary liver abscess caused by Klebsiella pneumoniae is an infection that is emerging worldwide and that is associated with severe morbidity and considerable mortality. A retrospective analysis of 110 episodes of primary liver abscess caused by K. pneumoniae that required hospitalization during 2001-2002 was conducted to identify predictors of metastatic infection, mortality, and the efficacy of first-generation cephalosporins and percutaneous drainage. The potential role of Klebsiella rmpA and magA genes was also evaluated. The study included 59 men and 51 women, with a mean age of 61.8 years. Diabetes was noted in 67 patients (60.9%). Metastatic infection occurred in 17 patients (15.5%), with meningitis accounting for 11 patients (64.7%) and endophthalmitis accounting for 4 patients (23.5%). The overall mortality rate was 10.0% (11 patients). Most of the severe complications occurred within the first 3 days after hospital admission. Ninety-two patients (83.6%) received treatment with cefazolin for >3 days. Four patients (4.3%) of the group who received cefazolin had metastatic infection, 1 patient (1.1%) experienced septic shock, and 3 (3.3%) experienced acute respiratory failure. Five (5.4%) of those 92 patients died. Multivariable analysis revealed that rmpA (odds ratio [OR], 28.85), Acute Physiologic and Chronic Health Evaluation (APACHE) II score >or=20 (OR, 8.08), and septic shock (OR, 4.33) were statistically significant predictors of metastatic infection. Metastatic infection (OR, 6.73), severity of disease (APACHE II score >or=16; OR, 11.82), septic shock (OR, 8.30), acute respiratory failure (OR, 69.92), and gas formation revealed on imaging (OR, 13.26) predicted mortality. Pigtail drainage protected against both metastatic infection (OR, 0.25) and mortality (OR, 0.14). Management of primary liver abscess caused by K. pneumoniae with use of first-generation cephalosporins and percutaneous drainage was associated with low rates of mortality, metastatic infection, and complications. These rates are comparable to those reported for third-generation cephalosporins.


MicroRNA-34a induces endothelial progenitor cell senescence and impedes its angiogenesis via suppressing silent information regulator 1.
Endothelial progenitor cells (EPCs) play an important role in angiogenesis, which is essential for numerous physiological processes as well as tumor growth. Several microRNAs (miRNAs) have been reported to be involved in angiogenesis. MiR-34a, recently reported as a tumor suppressor, has been found to target silent information regulator 1 (Sirt1), leading to cell cycle arrest or apoptosis. However, the role of miR-34a in EPC-mediated angiogenesis was unknown. The present study tested the hypothesis that miR-34a inhibits EPC-mediated angiogenesis by inducing senescence via suppressing Sirt1. Bone marrow-derived EPCs from adult male Sprague-Dawley rats were used. Results of flow cytometry showed that EPCs after 7 days of culture expressed both stem cell markers CD34 and CD133 and endothelial cell markers VEGFR-2 (flk-1) and VE-cadherin. MiR-34a was expressed in normal EPCs, and overexpression of miR-34a via its mimic transfection significantly increased its expression and impaired in vitro EPC angiogenesis. MiR-34a overexpression led to a significantly increased EPC senescence, paralleled with an approximately 40% Sirt1 reduction. Furthermore, knockdown of Sirt1 by its siRNA resulted in diminished EPC angiogenesis and increased senescence. Finally, overexpression of miR-34a increased the level of Sirt1 effector-acetylated forkhead box O transcription factors 1 (FoxO1), an effect mimicked in EPCs following Sirt1 knockdown. In conclusion, miR-34a impairs EPC-mediated angiogenesis by induction of senescence via inhibiting Sirt1.


Measurement of axial forces during emptying from the human stomach.
Our aim was to measure axial forces in the stomach and to evaluate their relation to circumferential contractions of the gastric walls and the emptying of gastric content. We used a combination of simultaneous radioscintigraphy, gastroduodenal manometry, and an axial force transducer with an inflatable 2-ml balloon fluoroscopically placed in the antrum. In vitro studies demonstrated that the axial force transducer records only antegrade forces along the longitudinal axis of this probe in an intensity-dependent manner. In vivo studies were performed in five healthy subjects for at least 3 h after ingestion of radiolabeled meals. When administered separately, the emptying of liquids or solids from the stomach is associated with generation of antral axial forces and coincident phasic pressure activity; however, almost 20% (average) of gastric axial forces during emptying of liquids or solids are unassociated with proximal or distal antral pressure activity ("isolated" forces). High amplitude antral axial forces and pressures occur during both lag and postlag emptying phases. During emptying of liquids, there is a trend for axial forces to be coincident more often with proximal than with distal antral pressure activity and vice versa for the emptying of solids (P = 0.015). These data suggest that when placed in the antrum, the transducer can semiquantitatively record axial forces during gastric emptying. By combining these observations with the data from in vitro studies, it appears that axial forces predominantly result from traction on the balloon by the longitudinal vector resulting from circumferential gastric contractions. The combination of radioscintigraphy and measurement of antral axial forces is a promising method to evaluate mechanical forces involved in the emptying of the human stomach.


The impact of visual layout factors on performance in Web pages: a cross-language study.
Visual layout has a strong impact on performance and is a critical factor in the design of graphical user interfaces (GUIs) and Web pages. Many design guidelines employed in Web page design were inherited from human performance literature and GUI design studies and practices. However, few studies have investigated the more specific patterns of performance with Web pages that may reflect some differences between Web page and GUI design. We investigated interactions among four visual layout factors in Web page design (quantity of links, alignment, grouping indications, and density) in two experiments: one with pages in Hebrew, entailing right-to-left reading, and the other with English pages, entailing left-to-right reading. Some performance patterns (measured by search times and eye movements) were similar between languages. Performance was particularly poor in pages with many links and variable densities, but it improved with the presence of uniform density. Alignment was not shown to be a performance-enhancing factor. The findings are discussed in terms of the similarities and differences in the impact of layout factors between GUIs and Web pages. Actual or potential applications of this research include specific guidelines for Web page design.


Photoinitiators: a food safety review.
Ultraviolet light printing inks are considered safer than the classical inks; however, despite being on the outer surface of the packaging material, their components can migrate into foodstuffs and can give rise to contamination. Photoinitiators are a part of the formulation of printing inks, being an important class of migrant, for which there have been more than 100 incidents of contamination of packaged food with photoinitiators reported through Rapid Alert System for Food and Feed (RASFF) alerts in the European Union. In this review the process of photo-polymerisation is explained in depth to provide an insight into the complexity of the process, and the diversity of potential contaminants together with their degradation products. The critical factors affecting the migration process itself are reviewed, together with analytical methods and the current legislation in the European Union and other parts of the world.


Antihypertensive drug classes and the risk of hip fracture: results from the Swedish primary care cardiovascular database.
Hypertension and fractures related to osteoporosis are major public health problems that often coexist. This study examined the associations between exposure to different antihypertensive drug classes and the risk of hip fracture in hypertensive patients. We included 59 246 individuals, 50 years and older, diagnosed with hypertension during 2001-2008 in the Swedish Primary Care Cardiovascular Database. Patients were followed from 1 January 2006 (or the date of diagnosis of hypertension) until they had their first hip fracture, died, or reached the end of the study on 31 December 2012. Cox proportional hazards models were used to calculate the risk of hip fracture across types of antihypertensive medications, adjusted for age, sex, comorbidity, medications, and socioeconomic factors. In total, 2593 hip fractures occurred. Compared to nonusers, current use of bendroflumethiazide or hydrochlorothiazide was associated with a reduced risk of hip fracture (hazard ratio 0.86; 95% CI 0.75-0.98 and hazard ratio 0.84; 95% CI 0.74-0.96, respectively), as was use of fixed drug combinations containing a thiazide (hazard ratio 0.69; 95% CI 0.57-0.83). Current use of loop diuretics was associated with an increased risk of hip fracture (hazard ratio 1.23; 95% CI 1.11-1.35). No significant associations were found between the risk of hip fracture and current exposure to beta-blockers, angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, aldosterone-receptor blockers or calcium channel blockers. In this large observational study of hypertensive patients, the risk of hip fracture differed across users of different antihypertensive drugs, results that could have practical implications when choosing antihypertensive drug therapy.


Eliciting views on antibiotic prescribing and resistance among hospital and outpatient care physicians in Berlin, Germany: results of a qualitative study.
To better understand physicians' views on factors of influence for the prescribing of antibiotics and on antibiotic resistance in the Berlin region, Germany. Qualitative study with focus groups. Outpatient care and hospital care practice in the Berlin region, Germany. 7 General practitioners, two urologists, one paediatrician from outpatient care and eight internists, two paediatricians, two ear, nose and throat specialists and two urologists from hospital care. Physicians showed differential interest in topics related to antibiotic prescribing and antibiotic resistance. Outpatient care physicians were interested in topics around their own prescribing, such as being able to diagnose and prescribe precisely, and topics about patient demand and non-compliance. Hospital care physicians were interested in hygiene challenges, limited consult time and multi-resistant pathogens. Physicians considered the development of resistance to be more in the domain of clinical treatment than that of the patient. Major challenges related to antibiotic resistance for this group of physicians are access to and clarity of treatment recommendations, implementation of hygienic measures, as well as increased outsourcing of laboratory services. Results raise questions about whether meeting physicians' expectations should be a focus when developing intervention that aims to influence antibiotic resistance in this and other areas of Germany.


Initial, habitual and compulsive alcohol use is characterized by a shift of cue processing from ventral to dorsal striatum.
During the development of drug addiction, initial hedonic effects decrease when substance use becomes habitual and ultimately compulsive. Animal research suggests that these changes are represented by a transition from prefrontal cortical control to subcortical striatal control and within the striatum from ventral to dorsal domains of the striatum, but only limited evidence exists in humans. In this study we address this hypothesis in the context of alcohol dependence. Non-abstinent heavy social drinkers (n = 21, 5.0 ± 1.5 drinks/day, 13 of them were alcohol-dependent according to DSM-IV) and light social drinkers (n = 10, 0.4 ± 0.4 drinks/day) were examined. We used a cue-reactivity functional magnetic resonance imaging (fMRI) design during which pictures of alcoholic beverages and neutral control stimuli were presented. In the dorsal striatum heavy drinkers showed significant higher activations compared to light drinkers, whereas light social drinkers showed higher cue-induced fMRI activations in the ventral striatum and in prefrontal areas compared to heavy social drinkers [region of interest analyses, P < 0.05 false discovery rate (FDR)-corrected]. Correspondingly, ventral striatal activation in heavy drinkers correlated negatively with obsessive-compulsive craving, and furthermore we found a positive association between cue-induced activation in the dorsal striatum and obsessive-compulsive craving in all participants. In line with our hypothesis we found higher cue-induced activation of the ventral striatum in social compared to heavy drinkers, and higher dorsal striatal activation in heavy drinkers. Increased prefrontal activation may indicate that social drinkers activate cortical control when viewing alcohol cues, which may prevent the development of heavy drinking or alcohol dependence. Our results suggest differentiating treatment research depending on whether alcohol use is hedonic or compulsive.


Molecular dynamics approach to understand the denaturing effect of a millimolar concentration of dodine on a λ-repressor and counteraction by trehalose.
Commonly used denaturants for protein denaturation are conventionally required in high concentrations in order to produce their effects. In this study, a molecular dynamics simulation of a mutated version of the N-terminal domain of a λ-repressor is carried out in the presence of a 10 millimolar (mM) concentration of dodine. Such a small concentration is found to be effective for denaturation of the protein. Both electrostatic and van der Waals interactions (between protein and dodine) play a role in the denaturation process and we observe more denaturation at the terminal helices. Three different molar concentrations of trehalose are used in order to check the counteraction against the action of dodine. This study shows that 0.5 and 1.0 M trehalose are sufficient to counteract the action of dodine. The study also sheds light on the fact that some protein sites are more responsive to unfolding, which is evident from the helical fractions of the terminal helices for different systems. The counteraction of trehalose on dodine-induced protein denaturation is found to be due to the replacement of some of the dodine molecules by trehalose molecules in the solvation shell of the protein. Preferential solvation of dodine molecules by trehalose molecules through hydrogen bonding interactions also plays a vital role in stabilizing the native conformation of the protein in a high trehalose concentration. Replacement of protein-dodine and protein-water hydrogen bonds by protein-trehalose hydrogen bonds is also observed.


Foreign-born emigration: a new approach and estimates based on matched CPS files.
The utility of postcensal population estimates depends on the adequate measurement of four major components of demographic change: fertility, mortality, immigration, and emigration. Of the four components, emigration, especially of the foreign-born, has proved the most difficult to gauge. Without "direct" methods (i.e., methods identifying who emigrates and when), demographers have relied on indirect approaches, such as residual methods. Residual estimates, however are sensitive to inaccuracies in their constituent parts and are particularly ill-suited for measuring the emigration of recent arrivals. Here we introduce a new method for estimating foreign-born emigration that takes advantage of the sample design of the Current Population Survey (CPS): repeated interviews of persons in the same housing units over a period of 16 months. Individuals appearing in a first March Supplement to the CPS but not the next include those who died in the intervening year, those who moved within the country, and those who emigrated. We use statistical methods to estimate the proportion of emigrants among those not present in the follow-up interview. Our method produces emigration estimates that are comparable to those from residual methods in the case of longer-term residents (immigrants who arrived more than 10 years ago), but yields higher--and what appear to be more accurate--estimates for recent arrivals. Although somewhat constrained by sample size, we also generate estimates by age, sex, region of birth, and duration of residence in the United States.


TDP-43 induces mitochondrial damage and activates the mitochondrial unfolded protein response.
Mutations in or dys-regulation of the TDP-43 gene have been associated with TDP-43 proteinopathy, a spectrum of neurodegenerative diseases including Frontotemporal Lobar Degeneration (FTLD) and Amyotrophic Lateral Sclerosis (ALS). The underlying molecular and cellular defects, however, remain unclear. Here, we report a systematic study combining analyses of patient brain samples with cellular and animal models for TDP-43 proteinopathy. Electron microscopy (EM) analyses of patient samples revealed prominent mitochondrial impairment, including abnormal cristae and a loss of cristae; these ultrastructural changes were consistently observed in both cellular and animal models of TDP-43 proteinopathy. In these models, increased TDP-43 expression induced mitochondrial dysfunction, including decreased mitochondrial membrane potential and elevated production of reactive oxygen species (ROS). TDP-43 expression suppressed mitochondrial complex I activity and reduced mitochondrial ATP synthesis. Importantly, TDP-43 activated the mitochondrial unfolded protein response (UPRmt) in both cellular and animal models. Down-regulating mitochondrial protease LonP1 increased mitochondrial TDP-43 levels and exacerbated TDP-43-induced mitochondrial damage as well as neurodegeneration. Together, our results demonstrate that TDP-43 induced mitochondrial impairment is a critical aspect in TDP-43 proteinopathy. Our work has not only uncovered a previously unknown role of LonP1 in regulating mitochondrial TDP-43 levels, but also advanced our understanding of the pathogenic mechanisms for TDP-43 proteinopathy. Our study suggests that blocking or reversing mitochondrial damage may provide a potential therapeutic approach to these devastating diseases.


Continuity of healthcare for headache patients: a problem of communication between headache specialists and general practitioners.
The continuous care of headache patients, from headache centres to general practice, is a managerial problem that is still unsolved in Italy. In fact, if on the one hand patients do not usually go to headache centres because of poor information, on the other hand, if they do, they do not find their general practitioner (GP) sufficiently prepared to continue the management. In Sicily we have formed a dense network of headache centres that we will try to link on the Internet to deal with the problem of poor patients information and poor specialist consultation. We also have faced the problem of the continuous care, trying to overcome "the difficulties of communication between specialists, GPs and patients" and "the difficulties of GPs in diagnostic work", by simple instruments like the Italian version of ID-Migraine, a simple three-item questionnaire.


Radiogenomics: the promise of personalized treatment in radiation oncology?
Significant variability exists in normal tissue reactions in patients with cancer receiving radiotherapy, with a subpopulation exhibiting increased toxicity to ionizing radiation. Genomic studies have proposed that single nucleotide polymorphisms in DNA repair genes, cytokines, and reactive oxygen species may play a role in clinical radiosensitivity. Additional research examining the association between genetic variants and radiation-induced inflammation and fibrosis may spur the development of targeted therapy in radiation oncology, which could increase cure rates and limit toxicity. As more people become long-term cancer survivors, oncology nurses must aggressively assess and manage late treatment side effects to optimize patient functioning and quality of life. The purpose of the current article is to describe the effect of ionizing radiation on normal and irradiated tissue, discuss genetic mutations that are proposed to influence radiosensitivity, and identify future areas of research on the association between genetics and radiation toxicity.


Familial aggregation of arthritis-related diseases in seropositive and seronegative rheumatoid arthritis: a register-based case-control study in Sweden.
Our objective was to estimate the risk of developing rheumatoid arthritis (RA) associated with a family history of non-RA arthritis-related diseases. This familial co-aggregation is of clinical interest since it is often encountered when assessing family history of RA specifically, but also informative on the genetic overlap between these diseases. Since anticitrullinated peptide antibodies/rheumatoid factor (RF)-positive and RF-negative RA have both specific and shared genetic factors, the familial co-aggregation was assessed separately for seropositive and seronegative disease. Nested case-control study in prospectively recorded Swedish total population data. The Multi-Generation Register identified first-degree relatives. RA and arthritis-related diseases were ascertained through the nationwide patient register. RA serology was based on International Classification of Diseases tenth revision coded diagnoses, mainly reflecting RF. Familial risks were calculated using conditional logistic regression. Results were replicated using the Swedish rheumatology register. Familial co-aggregation was found between RA and every studied arthritis-related disease, but the magnitude varied widely, from juvenile idiopathic arthritis (JIA) (seropositive RA OR=3.98 (3.01 to 5.26); seronegative RA OR=5.70 (3.47 to 9.36)) to osteoarthritis (seropositive RA OR=1.03 (1.00 to 1.06); seronegative RA OR=1.05 (1.00 to 1.09)). The familial co-aggregation pattern of non-RA arthritis-related diseases was overall similar for seropositive and seronegative RA. Among those with family history of RA, relatives' other arthritis-related diseases conferred little or no additional risk. Although family history of several arthritis-related diseases may be useful to predict RA (eg, lupus and JIA), others (eg, osteoarthritis and arthralgia) are less useful. Seropositive and seronegative RA had rather similar familial co-aggregation patterns with arthritis-related diseases, suggesting that the two RA subsets are similar in the genetic factors that overlap with these diseases.


Random effects modelling versus logistic regression for the inclusion of cluster-level covariates in propensity score estimation: A Monte Carlo simulation and registry cohort analysis.
Purpose: Surgeon and hospital-related features, such as volume, can be associated with treatment choices and outcomes. Accounting for these covariates with propensity score (PS) analysis can be challenging due to the clustered nature of the data. We studied six different PS estimation strategies for clustered data using random effects modelling (REM) compared with logistic regression. Methods: Monte Carlo simulations were used to generate variable cluster-level confounding intensity [odds ratio (OR) = 1.01-2.5] and cluster size (20-1,000 patients per cluster). The following PS estimation strategies were compared: i) logistic regression omitting cluster-level confounders; ii) logistic regression including cluster-level confounders; iii) the same as ii) but including cross-level interactions; iv), v), and vi), similar to i), ii), and iii), respectively, but using REM instead of logistic regression. The same strategies were tested in a trial emulation of partial versus total knee replacement (TKR) surgery, where observational versus trial-based estimates were compared as a proxy for bias. Performance metrics included bias and mean square error (MSE). Results: In most simulated scenarios, logistic regression, including cluster-level confounders, led to the lowest bias and MSE, for example, with 50 clusters × 200 individuals and confounding intensity OR = 1.5, a relative bias of 10%, and MSE of 0.003 for (i) compared to 32% and 0.010 for (iv). The results from the trial emulation also gave similar trends. Conclusion: Logistic regression, including patient and surgeon-/hospital-level confounders, appears to be the preferred strategy for PS estimation.


How much do marine connectivity fluctuations matter?
There is growing awareness that fluctuations in larval flux (connectivity) depress the long-run growth of marine metapopulations, but by how much is unclear. Here, we explore how reproductive schedule and larval behavior affect how much connectivity fluctuations depress growth. We combine larval dispersal simulations from the Florida Keys with theoretical results to calculate the effect of fluctuations on bicolor damselfish (Stegastes partitus). We find that fluctuations depress growth only slightly (∼2%), but the effect would be much stronger for an organism that spawned only part of the year. Larval behavior can also matter, as vertical migration allows larvae to become entrained in eddies. Eddies synchronize connectivity fluctuations, further decreasing growth. However, here, they also divide the Keys into largely independently fluctuating regions, mitigating the effects of local synchrony. Therefore, in situations where connectivity fluctuations matter, the presence of independently fluctuating regions due to larval behavior may be important.


Towards Advanced Interactive Visualization for Virtual Atlases.
An atlas is generally defined as a bound collection of tables, charts or illustrations describing a phenomenon. In an anatomical atlas for example, a collection of representative illustrations and text describes anatomy for the purpose of communicating anatomical knowledge. The atlas serves as reference frame for comparing and integrating data from different sources by spatially or semantically relating collections of drawings, imaging data, and/or text. In the field of medical image processing, atlas information is often constructed from a collection of regions of interest, which are based on medical images that are annotated by domain experts. Such an atlas may be employed, for example, for automatic segmentation of medical imaging data. The combination of interactive visualization techniques with atlas information opens up new possibilities for content creation, curation, and navigation in virtual atlases. With interactive visualization of atlas information, students are able to inspect and explore anatomical atlases in ways that were not possible with the traditional method of presenting anatomical atlases in book format, such as viewing the illustrations from other viewpoints. With advanced interaction techniques, it becomes possible to query the data that forms the basis for the atlas, thus empowering researchers to access a wealth of information in new ways. So far, atlas-based visualization has been employed mainly for medical education, as well as biological research. In this survey, we provide an overview of current digital biomedical atlas tasks and applications and summarize relevant visualization techniques. We discuss recent approaches for providing next-generation visual interfaces to navigate atlas data that go beyond common text-based search and hierarchical lists. Finally, we reflect on open challenges and opportunities for the next steps in interactive atlas visualization.


Estimation of dynamic flux profiles from metabolic time series data.
Advances in modern high-throughput techniques of molecular biology have enabled top-down approaches for the estimation of parameter values in metabolic systems, based on time series data. Special among them is the recent method of dynamic flux estimation (DFE), which uses such data not only for parameter estimation but also for the identification of functional forms of the processes governing a metabolic system. DFE furthermore provides diagnostic tools for the evaluation of model validity and of the quality of a model fit beyond residual errors. Unfortunately, DFE works only when the data are more or less complete and the system contains as many independent fluxes as metabolites. These drawbacks may be ameliorated with other types of estimation and information. However, such supplementations incur their own limitations. In particular, assumptions must be made regarding the functional forms of some processes and detailed kinetic information must be available, in addition to the time series data. The authors propose here a systematic approach that supplements DFE and overcomes some of its shortcomings. Like DFE, the approach is model-free and requires only minimal assumptions. If sufficient time series data are available, the approach allows the determination of a subset of fluxes that enables the subsequent applicability of DFE to the rest of the flux system. The authors demonstrate the procedure with three artificial pathway systems exhibiting distinct characteristics and with actual data of the trehalose pathway in Saccharomyces cerevisiae. The results demonstrate that the proposed method successfully complements DFE under various situations and without a priori assumptions regarding the model representation. The proposed method also permits an examination of whether at all, to what degree, or within what range the available time series data can be validly represented in a particular functional format of a flux within a pathway system. Based on these results, further experiments may be designed to generate data points that genuinely add new information to the structure identification and parameter estimation tasks at hand.


A Random Forest-Induced Distance-Based Measure of Physiologic al Dysregulation.
Aging involves gradual, multisystemic Physiological Dysregulation (PD) which increases risk of age-related comorbidities. Ability to quantify age-related PD could provide insights into biological mechanisms underlying the aging process. One approach to measuring PD exploits the fact that increasing PD manifests as a gradual deviation of physiological parameters away from normal levels. A recent geometric approach for quantifying PD uses Mahalanobis distance to measure the extent to which an individual's physiological parameters (measured via biomarkers from clinical blood biochemistry panels) deviate from normal levels. While useful, this approach has shortcomings that may impact its accuracy, primarily the incorrect assumption of multivariate normality among biomarkers, and identical weighting of biomarkers. Herein, we develop a more robust multivariate distance-based measure of PD. Proximity matrices induced by survival tree ensembles (Random Survival Forests) were used to compute a robust distance metric for quantifying how abnormal an individual's biomarker profile is. This approach requires no distributional assumptions and allows differential weighting of biomarkers based on association with mortality. Using receiver operating characteristic analysis and model fit statistics we compared performance of our measure to the standard approach based on Mahalanobis distance. Our new metric showed statistically significant improvements in predicting mortality, health status and biological age, compared to the standard approach. Additional advantages offered by our method are the ability to handle missing values in biomarkers and to accommodate categorical risk factors. These results suggest our approach could provide greater precision in the evaluation of PD, which could enable better characterization of the extent and impact of degenerative processes resulting from aging.


Genetic Associations with Gestational Duration and Spontaneous Preterm Birth.
Despite evidence that genetic factors contribute to the duration of gestation and the risk of preterm birth, robust associations with genetic variants have not been identified. We used large data sets that included the gestational duration to determine possible genetic associations. We performed a genomewide association study in a discovery set of samples obtained from 43,568 women of European ancestry using gestational duration as a continuous trait and term or preterm (<37 weeks) birth as a dichotomous outcome. We used samples from three Nordic data sets (involving a total of 8643 women) to test for replication of genomic loci that had significant genomewide association (P<5.0×10-8) or an association with suggestive significance (P<1.0×10-6) in the discovery set. In the discovery and replication data sets, four loci (EBF1, EEFSEC, AGTR2, and WNT4) were significantly associated with gestational duration. Functional analysis showed that an implicated variant in WNT4 alters the binding of the estrogen receptor. The association between variants in ADCY5 and RAP2C and gestational duration had suggestive significance in the discovery set and significant evidence of association in the replication sets; these variants also showed genomewide significance in a joint analysis. Common variants in EBF1, EEFSEC, and AGTR2 showed association with preterm birth with genomewide significance. An analysis of mother-infant dyads suggested that these variants act at the level of the maternal genome. In this genomewide association study, we found that variants at the EBF1, EEFSEC, AGTR2, WNT4, ADCY5, and RAP2C loci were associated with gestational duration and variants at the EBF1, EEFSEC, and AGTR2 loci with preterm birth. Previously established roles of these genes in uterine development, maternal nutrition, and vascular control support their mechanistic involvement. (Funded by the March of Dimes and others.).


Methoxymethane C-O Bond Strengths: Do Their Changes Result from Hyperconjugation or Polar Effects?
The methoxymethanes have been studied and compared with the fluoromethanes. The energies and atomic charges were calculated using MP2/aug-cc-pVTZ, and the group separation energies and bond dissociation enthalpies were calculated using CBS-QB3. The group separation energies are endothermic and the BDE increases with additional substitution as a result of the increase in charge at the central carbon. The greater charge leads to a stronger bond to the new substituent as well as to the original substituents. With the methoxymethanes, there is a linear relationship between the BDE and the atomic charge at C. The energies of the several methoxymethane conformers were calculated, and their energies usually increase with increasing values of the electronic spatial extent in accord with a proposal by Gillespie. The role of hyperconjugation in these cases is not settled.


Anti-Inflammatory Effects of IL-27 in Zymosan-Induced Peritonitis: Inhibition of Neutrophil Recruitment Partially Explained by Impaired Mobilization from Bone Marrow and Reduced Chemokine Levels.
Rapid activation of the innate immune system is critical for an efficient host response to invading pathogens. However, the inflammatory reaction has to be strictly controlled to minimize harmful immunopathology. A number of mediators including the cytokine interleukin-27 (IL-27) appear to be responsible for limitation and resolution of inflammation. Despite increasing knowledge of its suppressive effects on T cells, the influence on neutrophils and macrophages is poorly understood. To determine the role of IL-27 in innate immune responses we analysed the effect of IL-27 in a T cell independent model of zymosan-induced peritonitis. Early administration of recombinant IL-27 strongly reduced the number of neutrophils recruited to the peritoneal cavity after zymosan application as well as the neutrophil frequency in the blood. Simultaneously, IL-27 reduced the release of neutrophils from the bone marrow upon inflammation. Although cytokine levels were not affected by IL-27 treatment, the levels of the chemokines KC, MCP-1 and MIP-1α in the peritoneal fluid were strongly decreased. These findings demonstrate that IL-27 is able to control mobilisation and recruitment of neutrophils into the peritoneal cavity and identify a novel mechanism to limit inflammation caused by innate immune cells.


Aberration-free three-dimensional multiphoton imaging of neuronal activity at kHz rates.
Multiphoton microscopy is a powerful tool in neuroscience, promising to deliver important data on the spatiotemporal activity within individual neurons as well as in networks of neurons. A major limitation of current technologies is the relatively slow scan rates along the z direction compared to the kHz rates obtainable in the x and y directions. Here, we describe a custom-built microscope system based on an architecture that allows kHz scan rates over hundreds of microns in all three dimensions without introducing aberration. We further demonstrate how this high-speed 3D multiphoton imaging system can be used to study neuronal activity at millisecond resolution at the subcellular as well as the population level.


Social phobia in Swedish adolescents : prevalence and gender differences.
The aim of this study was to investigate the prevalence of self-reported social phobia in a community sample of Swedish adolescents in junior high school, at the risk-period for developing social phobia. Of particular interest was to investigate gender differences in prevalence across ages. Prevalence of sub-threshold social phobia was also studied. Students in grades 6-8 (aged 12-14) from seventeen schools in five Swedish municipalities were screened by means of a self-report questionnaire, the social phobia screening questionnaire-for children (SPSQ-C). Data from a sample of 2,128 students were analysed and showed a point-prevalence rate of 4.4% (95%CI 3.5-5.2) and a significant gender difference (6.6% girls vs. 1.8% boys, P < 0.001). No significant differences in prevalence of probable cases emerged across the ages. At sub-threshold level, marked social fear of at least one social situation was reported by 13.8% of the total group. "Speaking in front of class" and "calling someone unfamiliar on the phone" were the most feared social situations. In the social phobia group, 91.4% reported impairment in the school-domain due to their social fear. Social phobia is a common psychiatric condition in Swedish adolescents, especially in girls. As impairment in the school-domain is reported to a high degree, professionals and teachers need to recognize social phobia in adolescents so that help in overcoming the difficulties can be offered.


Recurrent symbiont recruitment from fungal parasites in cicadas.
Diverse insects are associated with ancient bacterial symbionts, whose genomes have often suffered drastic reduction and degeneration. In extreme cases, such symbiont genomes seem almost unable to sustain the basic cellular functioning, which comprises an open question in the evolution of symbiosis. Here, we report an insect group wherein an ancient symbiont lineage suffering massive genome erosion has experienced recurrent extinction and replacement by host-associated pathogenic microbes. Cicadas are associated with the ancient bacterial co-obligate symbionts Sulcia and Hodgkinia, whose streamlined genomes are specialized for synthesizing essential amino acids, thereby enabling the host to live on plant sap. However, our inspection of 24 Japanese cicada species revealed that while all species possessed Sulcia, only nine species retained Hodgkinia, and their genomes exhibited substantial structural instability. The remaining 15 species lacked Hodgkinia and instead harbored yeast-like fungal symbionts. Detailed phylogenetic analyses uncovered repeated Hodgkinia-fungus and fungus-fungus replacements in cicadas. The fungal symbionts were phylogenetically intermingled with cicada-parasitizing Ophiocordyceps fungi, identifying entomopathogenic origins of the fungal symbionts. Most fungal symbionts of cicadas were uncultivable, but the fungal symbiont of Meimuna opalifera was cultivable, possibly because it is at an early stage of fungal symbiont replacement. Genome sequencing of the fungal symbiont revealed its metabolic versatility, presumably capable of synthesizing almost all amino acids, vitamins, and other metabolites, which is more than sufficient to compensate for the Hodgkinia loss. These findings highlight a straightforward ecological and evolutionary connection between parasitism and symbiosis, which may provide an evolutionary trajectory to renovate deteriorated ancient symbiosis via pathogen domestication.


Ontogeny and disease responses of Langerhans-like cells in lymphoid tissues of salmonid fish.
The ontogeny and disease responses of Langerhans-like cells within lymphoid tissues of Atlantic salmon, Salmo salar, and rainbow trout, Oncorhynchus mykiss, were investigated. These cells were studied in situ with the use of two markers: the ultrastructural presence of Birbeck-like granules and immunohistochemistry with an antibody against human langerin/CD207 that cross-reacts with salmonid tissues. The appearance of Birbeck-like granules was observed in rainbow trout at 2 weeks post-hatch (PH) in the thymus and anterior kidney prior to the development of the spleen. Spleen first appeared at 3 weeks PH in both Atlantic salmon and rainbow trout, and Birbeck-like granules were observed within cells of the newly developed spleens. The cross-reactivity of langerin as seen by immunohistochemistry was not clearly observed in kidney and spleen until 9 weeks PH, when a strong cytoplasmic reaction was observed. To study langerin-positive cells in spleen and kidney during disease, microsporidial gill disease (MGD) in rainbow trout was used as a known disease model inducing a strong cell-mediated adaptive immune response. Langerin-positive cells in healthy fish were seen predominantly in the spleen, and only low numbers were present in the anterior kidney. During MGD, langerin-positive cell numbers were elevated in the anterior kidney and were significantly higher during 5, 6, and 10 weeks post-exposure (PE) compared with healthy control tissue. During MGD, the distribution of langerin-positive cells in the spleen and anterior kidney shifted from having significantly higher numbers of cells in the spleen than in the kidney in controls and at 1 and 4 weeks PE to having a similar distribution of the cells in the two organs at 2, 3, 5, and 6 weeks PE. By 10 weeks PE, significantly higher numbers of langerin-positive cells occurred in the anterior kidney compared with the spleen.


BADGE: a novel Bayesian model for accurate abundance quantification and differential analysis of RNA-Seq data.
Recent advances in RNA sequencing (RNA-Seq) technology have offered unprecedented scope and resolution for transcriptome analysis. However, precise quantification of mRNA abundance and identification of differentially expressed genes are complicated due to biological and technical variations in RNA-Seq data. We systematically study the variation in count data and dissect the sources of variation into between-sample variation and within-sample variation. A novel Bayesian framework is developed for joint estimate of gene level mRNA abundance and differential state, which models the intrinsic variability in RNA-Seq to improve the estimation. Specifically, a Poisson-Lognormal model is incorporated into the Bayesian framework to model within-sample variation; a Gamma-Gamma model is then used to model between-sample variation, which accounts for over-dispersion of read counts among multiple samples. Simulation studies, where sequencing counts are synthesized based on parameters learned from real datasets, have demonstrated the advantage of the proposed method in both quantification of mRNA abundance and identification of differentially expressed genes. Moreover, performance comparison on data from the Sequencing Quality Control (SEQC) Project with ERCC spike-in controls has shown that the proposed method outperforms existing RNA-Seq methods in differential analysis. Application on breast cancer dataset has further illustrated that the proposed Bayesian model can 'blindly' estimate sources of variation caused by sequencing biases. We have developed a novel Bayesian hierarchical approach to investigate within-sample and between-sample variations in RNA-Seq data. Simulation and real data applications have validated desirable performance of the proposed method. The software package is available at http://www.cbil.ece.vt.edu/software.htm.


Role of root hairs and lateral roots in silicon uptake by rice.
The rice plant (Oryza sativa L. cv Oochikara) is known to be a Si accumulator, but the mechanism responsible for the high uptake of Si by the roots is not well understood. We investigated the role of root hairs and lateral roots in the Si uptake using two mutants of rice, one defective in the formation of root hairs (RH2) and another in that of lateral roots (RM109). Uptake experiments with nutrient solution during both a short term (up to 12 h) and relatively long term (26 d) showed that there was no significant difference in Si uptake between RH2 and the wild type (WT), whereas the Si uptake of RM109 was much less than that of WT. The number of silica bodies formed on the third leaf in RH2 was similar to that in WT, but the number of silica bodies in RM109 was only 40% of that in WT, when grown in soil amended with Si under flooded conditions. There was also no difference in the shoot Si concentration between WT and RH2 when grown in soil under upland conditions. Using a multi-compartment transport box, the Si uptake at the root tip (0-1 cm, without lateral roots and root hairs) was found to be similar in WT, RH2, and RM109. However, the Si uptake in the mature zone (1-4 cm from root tip) was significantly lower in RM109 than in WT, whereas no difference was found in Si uptake between WT and RH2. All these results clearly indicate that lateral roots contribute to the Si uptake in rice plant, whereas root hairs do not. Analysis of F(2) populations between RM109 and WT showed that Si uptake was correlated with the presence of lateral roots and that the gene controlling formation of lateral roots and Si uptake is a dominant gene.


Organic-inorganic-hybrid-polymer microlens arrays with tailored optical characteristics and multi-focal properties.
Plano-convex microlens arrays of organic-inorganic polymers with tailored optical properties are presented. The fine-tuning of each microlens within an array is achieved by confining inkjet printed drops of the polymeric ink onto pre-patterned substrates. The lens optical properties are thus freely specified, and high numerical apertures from 0.45 to 0.9 and focal lengths between 10 μm and 100 μm are demonstrated, confirming theoretical predictions. Combining nanoimprint lithography approaches and inkjet printing enables using the same material for the microlenses and their substrates, improving the optical performances. Microlens arrays with desired specifications are printed reaching yields up to 100% and high lens reproducibility with standard deviations of the apparent contact angle under 1° and of the numerical apertures and focal lengths under 6%. Microlens arrays involving lenses with different characteristics, e.g. multi focal length, and thus focal planes separated by only few microns are printed with the same reproducibility.


The effect of post traumatic stress disorders on rehabilitation among combat-wounded veterans.
In June 2008 the Congressionally Directed Medical Research Program provided a grant to the Research Foundation at the James A Haley Veterans Hospital in Tampa, Florida to provide care for wounded veterans from Operation Enduring Freedom (Afghanistan) and Operation Iraqi Freedom (OEF/OIF). The telerehabilitation for OEF/OIF returnees with mild or moderate combat related Traumatic Brain Injury (TBI) has as its objectives 1) care coordination for wounded veterans using distance technology via the internet and 2) monitoring of physical and mental health outcomes using a variety of instruments. A total of 75 veterans were enrolled in the study. Our initial findings indicate that 1) Functional capabilities measured by locomotion and mobility appear to have stabilized among our cohort of veterans while deficiencies in cognition (memory, problem solving), psychosocial adjustment (anger, emotional status) and problems in integrating into society pose challenges 2) Those with comorbid PTSD appear to linger in employability and ultimate integration into society as compared to those without the diagnosis 3) Individualized treatment pathways are needed for rehabilitation and ultimate integration into society.


Risk factors of infant and child mortality in rural Burkina Faso.
The aim of the study was to quantify the effect of risk factors for childhood mortality in a typical rural setting in sub-Saharan Africa. We performed a survival analysis of births within a population under demographic surveillance from 1992 to 1999 based on data from a demographic surveillance system in 39 villages around Nouna, western Burkina Faso, with a total population of about 30000. All children born alive in the period 1 January 1993 to 31 December 1999 in the study area (n = 10 122) followed-up until 31 December 1999 were included. All-cause childhood mortality was used as outcome variable. Within the observation time, 1340 deaths were recorded. In a Cox regression model a simultaneous estimation of hazard rate ratios showed death of the mother and being a twin as the strongest risk factors for mortality. For both, the risk was most pronounced in infancy. Further factors associated with mortality include age of the mother, birth spacing, season of birth, village, ethnic group, and distance to the nearest health centre. Finally, there was an overall decrease in childhood mortality over the years 1993-99. The study supports the multi-causation of childhood deaths in rural West Africa during the 1990s and supports the overall trend, as observed in other studies, of decreasing childhood mortality in these populations. The observed correlation between the factors highlights the need for multivariate analysis to disentangle the separate effects. These findings illustrate the need for more comprehensive improvement of prenatal and postnatal care in rural sub-Saharan Africa.


Attenuation of cigarette smoke-induced airway mucus production by hydrogen-rich saline in rats.
Over-production of mucus is an important pathophysiological feature in chronic airway disease such as chronic obstructive pulmonary disease (COPD) and asthma. Cigarette smoking (CS) is the leading cause of COPD. Oxidative stress plays a key role in CS-induced airway abnormal mucus production. Hydrogen protected cells and tissues against oxidative damage by scavenging hydroxyl radicals. In the present study we investigated the effect of hydrogen on CS-induced mucus production in rats. Male Sprague-Dawley rats were divided into four groups: sham control, CS group, hydrogen-rich saline pretreatment group and hydrogen-rich saline control group. Lung morphology and tissue biochemical changes were determined by immunohistochemistry, Alcian Blue/periodic acid-Schiff staining, TUNEL, western blot and realtime RT-PCR. Hydrogen-rich saline pretreatment attenuated CS-induced mucus accumulation in the bronchiolar lumen, goblet cell hyperplasia, muc5ac over-expression and abnormal cell apoptosis in the airway epithelium as well as malondialdehyde increase in the BALF. The phosphorylation of EGFR at Tyr1068 and Nrf2 up-regulation expression in the rat lungs challenged by CS exposure were also abrogated by hydrogen-rich saline. Hydrogen-rich saline pretreatment ameliorated CS-induced airway mucus production and airway epithelium damage in rats. The protective role of hydrogen on CS-exposed rat lungs was achieved at least partly by its free radical scavenging ability. This is the first report to demonstrate that intraperitoneal administration of hydrogen-rich saline protected rat airways against CS damage and it could be promising in treating abnormal airway mucus production in COPD.


Structure, dynamics and thermodynamics of the human centrin 2/hSfi1 complex.
Centrin, an EF-hand calcium-binding protein, has been shown to be involved in the duplication of centrosomes, and Sfi1 (Suppressor of fermentation-induced loss of stress resistance protein 1) is one of its centrosomal targets. There are three isoforms of human centrin, but here we only considered centrin 2 (HsCen2). This protein has the ability to bind to any of the approximately 25 repeats of human Sfi1 (hSfi1) with more or less affinity. In this study, we mainly focused on the 17th repeat (R17-hSfi1-20), which presents the highest level of similarity with a well-studied 17-residue peptide (P17-XPC) from human xeroderma pigmentosum complementation group C protein, another centrin target for DNA repair. The only known structure of HsCen2 was resolved in complex with P17-XPC. The 20-residue peptide R17-hSfi1-20 exhibits the motif L8L4W1, which is the reverse of the XPC motif, W1L4L8. Consequently, the dipole of the helix formed by this motif has a reverse orientation. We wished to ascertain the impact of this reversal on the structure, dynamics and affinity of centrin. To address this question, we determined the structure of C-HsCen2 [the C-terminal domain of HsCen2 (T94-Y172)] in complex with R17-hSfi1-20 and monitored its dynamics by NMR, after having verified that the N-terminal domain of HsCen2 does not interact with the peptide. The structure shows that the binding mode is similar to that of P17-XPC. However, we observed a 2 -A translation of the R17-hSfi1-20 helix along its axis, inducing less anchorage in the protein and the disruption of a hydrogen bond between a tryptophan residue in the peptide and a well-conserved nearby glutamate in C-HsCen2. NMR dynamic studies of the complex strongly suggested the existence of an unusual calcium secondary binding mode in calcium-binding loop III, made possible by the uncommon residue composition of this loop. The secondary metal site is only populated at high calcium concentration and depends on the type of bound ligand.


Randomized Trial to Compare Renal Function and Ductal Response between Indomethacin and Ibuprofen Treatment in Extremely Low Birth Weight Infants.
A head to head comparison study on renal function and ductal response between indomethacin and ibuprofen has rarely been conducted in extremely low birth weight (ELBW) infants. The aim was to compare renal function and ductal response between indomethacin and ibuprofen in ELBW infants. We performed a double-blind randomized control trial to compare renal function and ductal response between indomethacin (0.2, 0.1, and 0.1 mg/kg i.v. every 24 h for 3 doses) and ibuprofen lysine (10, 5, and 5 mg/kg i.v. every 24 h for 3 doses) in ELBW infants with significant hemodynamic patent ductus arteriosus (cardiovascular dysfunction score >3 and LA/AO ratio ≥1.3). A total of 144 infants were enrolled: 73 received indomethacin and 71 received ibuprofen lysine. Significant decreases in urine output were seen in 30 infants (41%) in the indomethacin group and 15 (21%) in the ibuprofen group (p = 0.02). The indomethacin group was associated with a significantly higher chance of persistent ductal response than the ibuprofen group (66 vs. 49%, p = 0.046), but with a lower glomerular filtration rate on day 1, higher serum creatinine on days 1, 2, and 7, and lower urinary prostaglandin on days 2-7. Both groups were comparable in mortality and in bronchopulmonary dysplasia, intraventricular hemorrhage, necrotizing enterocolitis, and retinopathy of prematurity morbidity. With the current dosage, ibuprofen had fewer renal side effects but was associated with a lower rate of persistent ductal closure in ELBW infants. The precise role of prostaglandin on renal tubular function in ELBW infants remains to be further investigated.


Enhancement of second-harmonic generation from self-assembled monolayers on gold by excitation with a radially polarized beam.
We have studied the enhancement of second-harmonic generation (SHG) from self-assembled monolayers on Au surfaces excited by radially polarized beams. The electric field at the metal surface was enhanced by constructive interference between the incident and the reflected beams due to a longitudinal field, which is the field parallel to the optical axis, generated around the focus by the radially polarized beam. Since even-order nonlinear phenomena are surface sensitive, the combination of SHG and a radially polarized beam has the potential to be a powerful new imaging tool for characterization of metal surfaces. The SHG signal excited by the radially polarized beam was about 3 times higher than that excited by a linearly polarized beam; in addition, the SHG from a 7-(dimethylamino)-4-methylcoumarin-3-isothiocyanate monolayer was about 1.3 times higher than that from a bare Au substrate.


The effect of lifting during work on low back pain: a health impact assessment based on a meta-analysis.
Lifting at work is considered an important risk factor for low back pain (LBP). However, contradictory findings have been reported, partly because frequency, duration and intensity (ie, the weight of the load) of lifting have not been systematically considered. This has hampered developments of threshold values for lifting. The aims of this study were: to assess the effect of lifting during work (quantified in duration, frequency or intensity) on the incidence of LBP and to quantify the impact of these relationships on the occurrence of LBP in occupational populations exposed to lifting. We searched in PubMed and EMBASE.com for longitudinal studies assessing the effect of occupational lifting on LBP incidence. For each study, the exposure-response slope of the association was estimated by loglinear regression analysis. When possible, a meta-analysis on these slopes was conducted. In a health impact assessment, the effects of the pooled exposure-response relationships on LBP incidence was assessed. Eight longitudinal studies were included. Pooled estimates resulted in ORs of 1.11 (1.05 to 1.18) per 10 kg lifted and 1.09 (1.03 to 1.15) per 10 lifts/day. Duration of lifting could not be pooled. Using these ORs, we estimated that lifting loads over 25 kg and lifting at a frequency of over 25 lifts/day will increase the annual incidence of LBP by 4.32% and 3.50%, respectively, compared to the incidence of not being exposed to lifting. Intensity and frequency of lifting significantly predict the occurrence of LBP. Exposure-response relationships show that lifting heavy loads may have a substantial impact on musculoskeletal health of the working population. This information may direct the development of occupational lifting guidelines and workplace design for LBP prevention.


[Developing Perceived Competence Scale (PCS) for Adolescents].
In this study, Perceived Competence Scale was developed to measure high school students' perceived competence. Scale development process was verified on three different samples. Participants of the research are some high school students in 2011-2012 academic terms from Ankara. Participants' numbers are incorporated in exploratory factor analysis, confirmatory factor analysis and test-retest reliability respectively, as follows: 372, 668 and 75. Internal consistency coefficients (Cronbach's and stratified α) are calculated separately for each group. For data analysis Factor 8.02 and LISREL 8.70 package programs were used. According to results of the analyses, internal consistency coefficients (α) are .90 - .93 for academic competence, .82 - .86 for social competence in the samples that exploratory and confirmatory factor analysis performed. For the whole scale internal consistency coefficient (stratified α) is calculated as .91. As a result of test-retest reliability, adjusted correlation coefficients (r) are .94 for social competence and .90 for academic competence. In addition, to fit indexes and regression weights obtained from factor analysis, findings related convergent and discriminant validity, indicating that competence can be addressed in two dimensions which are academic (16 items) and social (14 items).


Improved sensitivity of BCR-ABL detection: a triple-probe three-color fluorescence in situ hybridization system.
Chronic myeloid leukemia is a clonal stem cell disorder associated with the Philadelphia (Ph) translocation [t(9;22) (q34;q11)]. As a result of the Ph translocation, parts of the ABL and BCR genes become fused. Cytogenetic quantification of Ph+ metaphases can be used to monitor patient response to treatment but is of limited sensitivity and applies only to cycling cells. Fluorescence in situ hybridization (FISH) with probes from the BCR and ABL regions can also identify the Ph translocation in interphase cells. Established systems for the detection of fusion genes by FISH rely on colocalization of two different probes but are associated with a high rate of false-positive results. We have introduced a third probe labeled with a different fluorochrome to create a triple-probe/three-color system that permits identification of both the Ph chromosome and the derivative 9 chromosome in Ph+ cells. This system was used to determine the frequency of interphase cells carrying the BCR-ABL fusion gene in bone marrow and peripheral blood granulocytes from patients showing variable cytogenetic responses to interferon. Our data show that the triple-probe/three-color approach allows highly sensitive detection of residual disease. Moreover, this method is readily applicable to the analysis of other chromosome translocations.


Conical intersections and semiclassical trajectories: comparison to accurate quantum dynamics and analyses of the trajectories.
Semiclassical trajectory methods are tested for electronically nonadiabatic systems with conical intersections. Five triatomic model systems are presented, and each system features two electronic states that intersect via a seam of conical intersections (CIs). Fully converged, full-dimensional quantum mechanical scattering calculations are carried out for all five systems at energies that allow for electronic de-excitation via the seam of CIs. Several semiclassical trajectory methods are tested against the accurate quantum mechanical results. For four of the five model systems, the diabatic representation is the preferred (most accurate) representation for semiclassical trajectories, as correctly predicted by the Calaveras County criterion. Four surface hopping methods are tested and have overall relative errors of 40%-60%. The semiclassical Ehrenfest method has an overall error of 66%, and the self-consistent decay of mixing (SCDM) and coherent switches with decay of mixing (CSDM) methods are the most accurate methods overall with relative errors of approximately 32%. Furthermore, the CSDM method is less representation dependent than both the SCDM and the surface hopping methods, making it the preferred semiclassical trajectory method. Finally, the behavior of semiclassical trajectories near conical intersections is discussed.


Real-world objects are more memorable than photographs of objects.
Research studies in psychology typically use two-dimensional (2D) images of objects as proxies for real-world three-dimensional (3D) stimuli. There are, however, a number of important differences between real objects and images that could influence cognition and behavior. Although human memory has been studied extensively, only a handful of studies have used real objects in the context of memory and virtually none have directly compared memory for real objects vs. their 2D counterparts. Here we examined whether or not episodic memory is influenced by the format in which objects are displayed. We conducted two experiments asking participants to freely recall, and to recognize, a set of 44 common household objects. Critically, the exemplars were displayed to observers in one of three viewing conditions: real-world objects, colored photographs, or black and white line drawings. Stimuli were closely matched across conditions for size, orientation, and illumination. Surprisingly, recall and recognition performance was significantly better for real objects compared to colored photographs or line drawings (for which memory performance was equivalent). We replicated this pattern in a second experiment comparing memory for real objects vs. color photos, when the stimuli were matched for viewing angle across conditions. Again, recall and recognition performance was significantly better for the real objects than matched color photos of the same items. Taken together, our data suggest that real objects are more memorable than pictorial stimuli. Our results highlight the importance of studying real-world object cognition and raise the potential for applied use in developing effective strategies for education, marketing, and further research on object-related cognition.


A longitudinal study of systemic inflammation and recovery of lean body mass among malnourished HIV-infected adults starting antiretroviral therapy in Tanzania and Zambia.
The effects of inflammation on nutritional rehabilitation after starting antiretroviral therapy (ART) are not well understood. We assessed the relationship between inflammation and body composition among patients enrolled in the Nutritional Support for African Adults Starting Antiretroviral therapy (NUSTART) trial in Tanzania and Zambia from 2011 to 2013. HIV-infected, ART-eligible adults with body mass index (BMI) of <18.5 kg/m(2) enrolled in the NUSTART trial were eligible for this study. Anthropometric and body composition data were collected at recruitment and 6 and 12 weeks post ART and C-reactive protein (CRP) was measured at recruitment and 6 weeks. The relationships between CRP and body composition were assessed using multiple regression. Of the 1815 trial participants, 838 (46%) had baseline and 6-week CRP measurements. Median age was 36 years, 55% were females and median CD4 count was 135 cells/μl. A one-log reduction in CRP at 6 weeks was associated with increased mid-upper arm circumference (0.45 cm; 95% CI: 0.30, 0.61), calf circumference (0.38 cm; 0.23, 0.54), waist circumference (0.98 cm; 0.59, 1.37), BMI (0.37 kg/m(2); 0.24, 0.50) and fat-free mass (0.58 kg; 0.26, 0.91), but not with fat mass (0.09 kg; -0.17, 0.34). Fat-free mass gains persisted at 12 weeks and were more closely associated with 6-week CRP values than with baseline values. Reduction in CRP shortly after ART initiation was associated with higher fat-free mass gains. Further studies are warranted to determine whether interventions to reduce systemic inflammation will enhance gains in fat-free mass.


First Report of Laurel Wilt, Caused by Raffaelea lauricola, on Sassafras (Sassafras albidum) in Alabama.
Laurel wilt, caused by Raffaelea lauricola, a fungal symbiont of the redbay ambrosia beetle, Xyleborus glabratus, is responsible for extensive mortality of native redbays (Persea borbonia and P. palustris) in the coastal plains of the southeastern United States (1). The wilt also affects the more widespread sassafras, Sassafras albidum, particularly in areas where diseased redbays are common and populations of X. glabratus are high. Because sassafras stems were thought to lack chemicals that are attractive to the beetle, and sassafras tends to be widely scattered in forests, it was believed that the advance of the laurel wilt epidemic front might slow once it reached the edge of the natural range of redbay, which is restricted to the coastal plains of the Gulf and Atlantic Coasts (2). In July and August of 2011, wilt-like symptoms (i.e., wilted and dead leaves, and streaks of black discoloration in the xylem) were observed on 1 to 10 sassafras trees (15 to 23 cm diameter; 6 to 9 m height) at each of three locations, which were approximately 6 km from one another in Marengo Co., Alabama. Samples of the discolored wood from five trees were plated on malt agar amended with cycloheximide and streptomycin (CSMA), and a fungus morphologically identical to R. lauricola was isolated from each tree (1). For confirmation, a portion of the large subunit (28S) of the rDNA region of three of the isolates was sequenced (3); in each case, the sequence matched exactly that of other isolates of R. lauricola (EU123077) from the United States. Symptomatic trees were found at all three sites when revisited in April 2012, and approximately 20 sassafras trees in various stages of wilt were observed at one location, where only one diseased tree had been noted in 2011. Bolts were cut from the main stem of a symptomatic tree, and eggs, larvae, and adults of X. glabratus were commonly found in tunnels, and R. lauricola was isolated from the discolored xylem. Three container-grown sassafras saplings (mean height 193 cm, mean diameter 2.1 cm at groundline) were inoculated as previously described (1) with conidia (~600,000) from an isolate of R. lauricola. Three additional sassafras saplings were inoculated with sterile, deionized water, and all plants were placed in a growth chamber at 25°C with a 15-h photoperiod. Inoculated plants began to exhibit wilt symptoms within 14 days, and at 30 days all inoculated plants were dead and xylem discoloration was observed. Control plants appeared healthy and did not exhibit xylem discoloration. Pieces of sapwood from 15 cm above the inoculation points were plated on CSMA, and R. lauricola was recovered from all wilted plants but not from control plants. This is the first record of laurel wilt in Alabama and is significant because the disease appears to be spreading on sassafras in an area where redbays have not been recorded (see http://www.floraofalabama.org ). The nearest previously documented case of laurel wilt is on redbay and sassafras in Jackson Co., Mississippi (4), approximately 160 km to the south. The exact source of the introduction of X. glabratus and R. lauricola into Marengo Co. is not known. The vector may have been transported into the area with storms, moved with infested firewood, or shipped with infested timber by companies that supply mills in the area. References: (1) S. Fraedrich et al. Plant Dis. 92:215, 2008. (2) J. Hanula et al. Econ. Ent. 101:1276, 2008. (3) T. Harrington et al. Mycotaxon 111:337, 2010. (4) J. Riggins et al. Plant Dis. 95:1479, 2011.


Implementation of a multidisciplinary team approach and fish oil emulsion administration in the management of infants with short bowel syndrome and parenteral nutrition-associated liver disease.
To describe the authors' experience with the implementation of a multidisciplinary approach and use of fish oil emulsion (FOE) in the management of infants with short bowel syndrome (SBS) and parenteral nutrition-associated liver disease (PNALD). Between August 2006 and June 2009, four cases of SBS and severe PNALD were managed by the team using specifically developed protocols. The FOE was initiated if serum direct bilirubin levels were ≥100 µmol/L. To quantify the degree of exposure to high serum direct bilirubin levels over time, the area under the curve (AUC) for each patient was calculated before and after initiation of FOE. Linear regression analyses were performed to evaluate correlations between the AUC, duration of cholestasis and initiation of FOE. All patients survived and no complications were observed during the study period. After the first patient, FOE was initiated progressively earlier, but poor correlation between the AUC before and after its introduction was observed (r(2)=0.41924). However, there was strong correlation between the duration of PNALD before FOE initiation and time to resolution (r(2)=0.72133): the earlier the FOE was initiated, the shorter the time to resolution. The authors report a positive experience with the implementation of a multidisciplinary approach and the use of FOE in infants with SBS and severe PNALD. The earlier the FOE was initiated during the cholestatic process, the shorter the time to resolution. The present study is a hypothesis generator, raising the question of whether an earlier introduction of this particular therapy can effectively shorten the cholestasis process in these patients.


Sex and dose-related differences in methylphenidate adolescent locomotor sensitization and effects on brain-derived neurotrophic factor.
This study analyzed repeated methylphenidate (MPH) administration and its effects on brain-derived neurotrophic factor (BDNF) in the dorsal striatum and nucleus accumbens of male and female adolescent rats. In Experiment 1, rats were administered intraperitoneal (ip) saline, 1, 3, or 5 mg/kg dose of MPH every second day from postnatal day (P)33-P49. Locomotor activity was analyzed for 10 min after each administration. Results revealed that the 1 mg/kg dose of MPH produced locomotor suppression, however, the 5 mg/kg dose of MPH produced locomotor sensitization and robust behavioral activation in females as compared to males. In Experiment 2, animals were administered ip saline or the 5 mg/kg dose of MPH using an identical regimen but a 30 min behavioral test was employed. Dorsal striatum and nucleus accumbens tissue was assayed for BDNF at P50. Females demonstrated sensitization to MPH and increased locomotor activation compared to males. Interestingly, females given MPH demonstrated a significant 42% decrease of striatal BDNF whereas males administered MPH demonstrated a significant 50.4% increase of striatal BDNF compared to controls. There were no effects on accumbal BDNF. This report demonstrates robust sex differences in the behavioral response, but sex-dependent changes in striatal BDNF in response to MPH in adolescence.


SAM-CAAM: A Concept for Acquiring Systematic Aircraft Measurements to Characterize Aerosol Air Masses.
A modest operational program of systematic aircraft measurements can resolve key satellite-aerosol-data-record limitations. Satellite observations provide frequent, global aerosol-amount maps, but offer only loose aerosol property constraints needed for climate and air quality applications. We define and illustrate the feasibility of flying an aircraft payload to measure key aerosol optical, microphysical, and chemical properties in situ. The flight program could characterize major aerosol air-mass types statistically, at a level-of-detail unobtainable from space. It would: (1) enhance satellite aerosol retrieval products with better climatology assumptions, and (2) improve translation between satellite-retrieved optical properties and species-specific aerosol mass and size simulated in climate models to assess aerosol forcing, its anthropogenic components, and other environmental impacts. As such, Systematic Aircraft Measurements to Characterize Aerosol Air Masses (SAM-CAAM) could add value to data records representing several decades of aerosol observations from space, improve aerosol constraints on climate modeling, help interrelate remote-sensing, in situ, and modeling aerosol-type definitions, and contribute to future satellite aerosol missions. Fifteen Required Variables are identified, and four Payload Options of increasing ambition are defined, to constrain these quantities. "Option C" could meet all the SAM-CAAM objectives with about 20 instruments, most of which have flown before, but never routinely several times per week, and never as a group. Aircraft integration, and approaches to data handling, payload support, and logistical considerations for a long-term, operational mission are discussed. SAM-CAAM is feasible because, for most aerosol sources and specified seasons, particle properties tend to be repeatable, even if aerosol loading varies.


Perioperative Pain and Addiction Interdisciplinary Network (PAIN): consensus recommendations for perioperative management of cannabis and cannabinoid-based medicine users by a modified Delphi process.
In many countries, liberalisation of the legislation regulating the use of cannabis has outpaced rigorous scientific studies, and a growing number of patients presenting for surgery consume cannabis regularly. Research to date suggests that cannabis can impact perioperative outcomes. We present recommendations obtained using a modified Delphi method for the perioperative care of cannabis-using patients. A steering committee was formed and a review of medical literature with respect to perioperative cannabis use was conducted. This was followed by the recruitment of a panel of 17 experts on the care of cannabis-consuming patients. Panellists were blinded to each other's participation and were provided with rater forms exploring the appropriateness of specific perioperative care elements. The completed rater forms were analysed for consensus. The expert panel was then unblinded and met to discuss the rater form analyses. Draft recommendations were then created and returned to the expert panel for further comment. The draft recommendations were also sent to four independent reviewers (a surgeon, a nurse practitioner, and two patients). The collected feedback was used to finalise the recommendations. The major recommendations obtained included emphasising the importance of eliciting a history of cannabis use, quantifying it, and ensuring contact with a cannabis authoriser (if one exists). Recommendations also included the consideration of perioperative cannabis weaning, additional postoperative nausea and vomiting prophylaxis, and additional attention to monitoring and maintaining anaesthetic depth. Postoperative recommendations included anticipating increased postoperative analgesic requirements and maintaining vigilance for cannabis withdrawal syndrome.


Cholinergic mesencephalic neurons are involved in gait and postural disorders in Parkinson disease.
Gait disorders and postural instability, which are commonly observed in elderly patients with Parkinson disease (PD), respond poorly to dopaminergic agents used to treat other parkinsonian symptoms. The brain structures underlying gait disorders and falls in PD and aging remain to be characterized. Using functional MRI in healthy human subjects, we have shown here that activity of the mesencephalic locomotor region (MLR), which is composed of the pedunculopontine nucleus (PPN) and the adjacent cuneiform nucleus, was modulated by the speed of imagined gait, with faster imagined gait activating a discrete cluster within the MLR. Furthermore, the presence of gait disorders in patients with PD and in aged monkeys rendered parkinsonian by MPTP intoxication correlated with loss of PPN cholinergic neurons. Bilateral lesioning of the cholinergic part of the PPN induced gait and postural deficits in nondopaminergic lesioned monkeys. Our data therefore reveal that the cholinergic neurons of the PPN play a central role in controlling gait and posture and represent a possible target for pharmacological treatment of gait disorders in PD.


Risk of arthropathy after the Bristow-Latarjet repair: a radiologic and clinical thirty-three to thirty-five years of follow-up of thirty-one shoulders.
Transfer of the coracoid (Bristow-Latarjet [B-L]) is used to stabilize anterior shoulder instability. We report the long-term results of our first 31 operations with this method. Thirty-six patients (mean age, 26.7 years) had a B-L repair from 1977 to 1979. Five patients died, and during 2012 to 2013, the remaining 31 shoulders had a follow-up with questionnaire, physical examination, Western Ontario Shoulder Instability Index, Subjective Shoulder Value, Subjective Assessment of Shoulder Function, subjective assessment of loss of motion, and radiologic imaging. One patient required revision surgery because of recurrence and another because of repeat dislocation. Six patients reported subluxations. Eighteen patients (58%) were very satisfied, and 13 (42%) were satisfied. The mean Western Ontario Shoulder Instability Index score (100 possible) was 85, and the median score was 93. According to Samilson-Prieto classification of arthropathy of the shoulder, 39% were classified as normal, 27% as mild, 23% as moderate, and 11% as severe. The classification of arthropathy varied with observers and radiologic views. Age younger than 22 years at the primary dislocation meant less arthropathy at follow-up (P = .045). The degree of arthropathy 33 to 35 years after the B-L repair seems to follow the natural history of shoulder dislocation with respect to arthropathic joint degeneration. Postoperative restriction of external rotation does not increase later arthropathy.


Femtosecond laser electronic excitation tagging for quantitative velocity imaging in air.
Time-accurate velocity measurements in unseeded air are made by tagging nitrogen with a femtosecond-duration laser pulse and monitoring the displacement of the molecules with a time-delayed, fast-gated camera. Centimeter-long lines are written through the focal region of a ∼1 mJ, 810 nm laser and are produced by nonlinear excitation and dissociation of nitrogen. Negligible heating is associated with this interaction. The emission arises from recombining nitrogen atoms and lasts for tens of microseconds in natural air. It falls into the 560 to 660 nm spectral region and consists of multiple spectral lines associated with first positive nitrogen transitions. The feasibility of this concept is demonstrated with lines written across a free jet, yielding instantaneous and averaged velocity profiles. The use of high-intensity femtosecond pulses for flow tagging allows the accurate determination of velocity profiles with a single laser system and camera.


Evaluation of fracture resistance of roots obturated with resilon and thermoplasticized gutta-percha: An in vitro study.
To investigate and compare the fracture resistance of resilon and realseal filled root canals with those that were obturated with thermoplasticized gutta-percha and AH-plus endodontic sealer. Eighty extracted human mandibular single-rooted premolars stored in 10% formalin were used in this study. They were prepared by using a crown-down technique, debrided and irrigated with NaOCl, EDTA, and normal saline and divided into four groups. Group I = Negative control, canals prepared but no obturation was done; Group II = Positive control obturated with dual cure composite resin; Group III = Obturated with thermoplasticized GP and AH-plus; Group IV = Obturated with resilon and realseal. All root specimens were stored for two weeks in 100% humidity to allow for complete setting of the sealer. Each specimen was mounted in acrylic in a polyvinyl ring and tested for fracture resistance with a universal testing machine. The loading fixture of the machine was mounted with its spherical tip aligned with the center of the access opening of each root. A vertical loading force was applied until it fractured the root. The force values were subjected to statistical analysis: Kruskall-Wallis H-test and Mann-Whitney U-test. Group comparison revealed that Group II (positive control) had significantly higher values of resistance to fracture as compared to all the other groups. While Group IV (resilon with realseal) had significantly higher values of resistance to fracture as compared to Group I (negative control). On the basis of our findings, it can be concluded that filling the root canals with resilon and realseal increased the in vitro resistance to fracture of single-canal extracted teeth. resilon and realseal demonstrated high fracture-resistance values and could be an alternative to the conventional gutta-percha.


Selenomethionine-substituted Thermus thermophilus cytochrome ba3: characterization of the CuA site by Se and Cu K-EXAFS.
We have designed a gene that encodes a polypeptide corresponding to amino acids 44-168 of the Thermus thermophilus cytochrome ba3 subunit II [Keightley et al. (1995) J. Biol. Chem. 270, 20345-20358]. The resulting ba3-CuAt10 protein separated into two fractions (A and B) during cation exchange chromatography which were demonstrated to differ only by N-terminal acetylation in fraction A. When the gene was expressed in an Escherichia coli strain that is auxotrophic for methionine and grown in the presence of selenomethionine (Se(Met)), the single methionine of the CuAt10 protein was quantitatively replaced with Se(Met). Native (S(Met)) and Se(Met)-substituted proteins were characterized by electrospray mass, optical absorption, and EPR spectroscopies and by electrochemical analysis; they were found to have substantially identical properties. The Se(Met)-containing protein was further characterized by Se and Cu K-EXAFS which revealed Cu-Se bond lengths of 2.55 A in the mixed-valence form and 2.52 A in the fully reduced form of CuA. Further analysis of the Se- and Cu-EXAFS spectra yielded the Se-S(thiolate) distances and thereby information on the Se-Cu-Cu and Se-Cu-S(thiolate) angles. An expanded EXAFS structural model is presented.


Molecular modeling calculations of HIV-1 reverse transcriptase nonnucleoside inhibitors: correlation of binding energy with biological activity for novel 2-aryl-substituted benzimidazole analogues.
The energies and physical descriptors for the binding of 20 novel 1-(2,6-difluorobenzyl)-2-(2,6-difluorophenyl)benzimidazole analogues (BPBIs) to HIV-1 reverse transcriptase (RT) have been determined using Monte Carlo (MC) simulations. The crystallographic structure of the lead compound, 1-(2,6-difluorobenzyl)-2-(2,6-difluorophenyl)-4-methylbenzimidazole, was used as a starting point to model the inhibitors in both the bound and the unbound states. The energy terms and physical descriptors obtained from the calculations were correlated with their respective experimental EC(50) values, resulting in an r(2) value of 0.70 and a root-mean-square deviation (rms) of 0.53 kcal/mol. The terms in the correlation include the change in total Coulombic energy and solvent-accessible surface area. Structural analysis of the data files from the BPBI calculations reveals that all of the analogues with good biological activity show the formation of a hydrogen bond between the ligand and the backbone nitrogen atom of lysine 103. By use of the structural results, two novel BPBI inhibitors have been designed and calculations have been carried out. The results show the formation of the desired hydrogen bonds, and the DeltaG(binding) values predict the compounds to be excellent RT inhibitors. Subsequent synthesis and biological activity testing of these analogues have shown the validity of the predictive calculations. If the BPBIs are modeled in a site constructed from the crystal coordinates of a member of another class of nonnucleoside inhibitors (the 4,5,6,7-tetrahydroimidazo[4,5,1-jk][1,4]benzodiazepine-2(1H)-thione and -one (TIBO) compounds), the correlation with the same terms drops slightly, giving an r(2) value of 0.61 with an associated root-mean-square value of 0.53 kcal/mol. Conversely, if the TIBO compounds are modeled in a site constructed from the BPBI complex crystal coordinates, a correlation can be obtained using the drug-protein interaction energy and change in the total number of hydrogen bonds, giving an r(2) value of 0.63. These are the same descriptors that were used for the TIBO compounds modeled in their own sites, where the r(2) value was 0.72. These data suggest that it may be possible, in some cases, to design novel inhibitors utilizing structural data from related, but not identical, inhibitors.


Hypoproliferative human lamina propria T cells retain the capacity to secrete lymphokines when stimulated via CD2/CD28 pathways.
Human lamina propria (LP) T cells exhibit a reduced proliferative capacity in response to antigen-specific stimulation. To investigate the functional state of such hypoproliferative T cells, we determined the capacity of LP T cells to produce lymphokines when stimulated by monoclonal antibodies that crosslink either the TCR/CD3 complex or accessory pathway molecules (CD2,CD28). We found that TCR/CD3-mediated proliferative responses of LP T cells were greatly diminished when compared to peripheral blood (PB) T cells, but were largely restored when cells were preincubated in IL-2. Despite their proliferative hyporesponsiveness, LP T cells (as compared to PB T cells) secreted equal amounts of IL-2 and increased amounts of IFN-gamma, IL-4 and IL-5; these increased cytokine responses were most evident when cells were stimulated via the accessory pathways. In further studies, purified CD4+ LP T cells were compared with purified CD4+/CD45RO+ PB T cells (i.e., the PB T cell subset they most resemble). LP T cells produced significantly more IFN-gamma and IL-5 but less IL-4 than their CD45RO+ PB counterparts. Overall, LP T cells are unresponsive T cells following stimulation via the TCR/CD3 pathway but nevertheless retain the capacity to produce increased levels of TH1 and TH2-type lymphokines following stimulation via the CD2/CD28 accessory pathway; thus, they are best classified as modified "anergic" T cells.


Optical control of calcium-regulated exocytosis.
Neurons signal to each other and to non-neuronal cells as those in muscle or glands, by means of the secretion of neurotransmitters at chemical synapses. In order to dissect the molecular mechanisms of neurotransmission, new methods for directly and reversibly triggering neurosecretion at the presynaptic terminal are necessary. Here we exploit the calcium permeability of the light-gated channel LiGluR in order to reversibly manipulate cytosolic calcium concentration, thus controlling calcium-regulated exocytosis. Bovine chromaffin cells expressing LiGluR were stimulated with light. Exocytic events were detected by amperometry or by whole-cell patch-clamp to quantify membrane capacitance and calcium influx. Amperometry reveals that optical stimulation consistently triggers exocytosis in chromaffin cells. Secretion of catecholamines can be adjusted between zero and several Hz by changing the wavelength of illumination. Differences in secretion efficacy are found between the activation of LiGluR and native voltage-gated calcium channels (VGCCs). Our results show that the distance between sites of calcium influx and vesicles ready to be released is longer when calcium influx is triggered by LiGluR instead of native VGCCs. LiGluR activation directly and reversibly increases the intracellular calcium concentration. Light-gated calcium influx allows for the first time to control calcium-regulated exocytosis without the need of applying depolarizing solutions or voltage clamping in chromaffin cells. LiGluR is a useful tool to study the secretory mechanisms and their spatiotemporal patterns in neurotransmission, and opens a window to study other calcium-dependent processes such as muscular contraction or cell migration.


Biological Markers for Pulpal Inflammation: A Systematic Review.
Pulpitis is mainly caused by an opportunistic infection of the pulp space with commensal oral microorganisms. Depending on the state of inflammation, different treatment regimes are currently advocated. Predictable vital pulp therapy depends on accurate determination of the pulpal status that will allow repair to occur. The role of several players of the host response in pulpitis is well documented: cytokines, proteases, inflammatory mediators, growth factors, antimicrobial peptides and others contribute to pulpal defense mechanisms; these factors may serve as biomarkers that indicate the status of the pulp. Therefore, the aim of this systematic review was to evaluate the presence of biomarkers in pulpitis. The electronic databases of MEDLINE, EMBASE, Scopus and other sources were searched for English and non-English articles published through February 2015. Two independent reviewers extracted information regarding study design, tissue or analyte used, outcome measures, results and conclusions for each article. The quality of the included studies was assessed using a modification of the Newcastle-Ottawa-Scale. From the initial 847 publications evaluated, a total of 57 articles were included in this review. In general, irreversible pulpitis was associated with different expression of various biomarkers compared to normal controls. These biomarkers were significantly expressed not only in pulp tissue, but also in gingival crevicular fluid that can be collected non-invasively, and in dentin fluid that can be analyzed without extirpating the entire pulpal tissue. Such data may then be used to accurately differentiate diseased from healthy pulp tissue. The interplay of pulpal biomarkers and their potential use for a more accurate and biologically based diagnostic tool in endodontics is envisaged.


Secretory pattern of inhibin during estrous cycle and pregnancy in African (Loxodonta africana) and Asian (Elephas maximus) elephants.
The ovary of female elephants has multiple corpora lutea (CL) during the estrous cycle and gestation. The previous reports clearly demonstrated that inhibin was secreted from lutein cells as well as granulosa cells of antral follicles in cyclic Asian elephants. The aim of this study is to investigate the inhibin secretion during the pregnancy in African and Asian elephants. Two African elephants and two Asian elephants were subjected to this study. Circulating levels of immunoreactive (ir-) inhibin and progesterone were measured by radioimmunoassay. Four pregnant periods of an African elephant and three pregnant periods of an Asian elephant were analyzed in this study. Circulating levels of ir-inhibin started to increase at 1 or 2 week before the ovulation and reached the peak level 3 or 4 weeks earlier than progesterone during the estrous cycle in both African and Asian elephants. After last luteal phase, the serum levels of ir-inhibin remained low throughout pregnancy in both an African and an Asian elephant. The mean levels of ir-inhibin during the pregnancy were lower than the luteal phase in the estrous cycle despite high progesterone levels were maintained throughout the pregnancy. These results strongly suggest that CL secrete a large amount of progesterone but not inhibin during the pregnancy in elephants.


Wavelength independence and interdevice variability of optical coherence tomography.
Comparison of nerve fiber layer (NFL) thickness and macular retinal thickness measurements in two different commercial optical coherence tomography (OCT) machines with the same and different super luminescent diode lasers (SLDs). Thirty eyes of 30 subjects were studied, with each eye scanned on two different OCT machines on the same day. Three 3.4-mm diameter circumpapillary NFL scans and six 6-mm radial scans centered on the fovea were obtained. Macular volumes were calculated from the central 3.45 mm of the 6 radial scans. The first study (identical SLDs of 850 nm) included 10 eyes of 10 subjects for mean NFL thickness and 6 eyes of 6 subjects for macular volume. There was no systematic difference between the measurements on the two machines for NFL (difference = 5.6 microm, standard error [SE] = 3.8, P= .18) or macular volume (difference = -0.003 mm3, SE = 0.064, P = .96). The second study (SLDs of 850 and 820 nm) included 20 eyes of 20 subjects. There was no significant difference between the two machines for mean NFL (difference = 3.4 microm, SE = 2.9, P= 0.26) or macular volume (difference = -.017 mm3, SE = 0.017, P= .33). For the smaller areas of the macular scan, several (including the outer ring) showed a significant systematic difference between measurements at the two wavelengths, but even these had at most 12% of the total variance attributable to the different wavelengths. Our study indicates that OCT measurements of NFL thickness do not differ much when the device or the wavelength of the SLD is changed. Some measurements of the macular thickness in specific areas may differ systematically for different wavelengths, but in most cases the difference is small.


The Role of Endocrine Disruptors in the Epigenetics of Reproductive Disease and Dysfunction: Potential Relevance to Humans.
In a murine model, we have linked early life toxicant exposure to reduced uterine sensitivity to progesterone, a phenotype we had previously associated with inflammation in endometriosis patients. Subsequent studies revealed that developmental toxicant exposure not only reduces fertility in male and female mice but also negatively impacts pregnancy leading to spontaneous preterm birth (PTB). An epigenetic alteration of the progesterone receptor gene correlated with reduced fertility and adverse pregnancy outcomes and persisted in multiple generations of mice in the absence of an additional toxicant exposure. Gene-environment interactions in women may explain why some patients "at risk" for PTB deliver at term while others without known risks deliver early. Our model provides a unique system to unravel the interactive influences of inflammation and reduced progesterone responsiveness on PTB and suggests that therapy needs to begin prior to pregnancy (and involve both partners) rather than once the inflammatory cascade has been initiated.