The impact performance of headguards for combat sports.
To assess the impact energy attenuation performance of a range of headguards for combat sports. Seven headguards worn during combat sport training or competition, including two Association Internationale de Boxe Amateur (AIBA)-approved boxing models, were tested using drop tests. An International Organization for Standardization (ISO) rigid headform was used with a 5.6 kg drop assembly mass. Tests were conducted against a flat rigid anvil both with and without a boxing glove section. The centre forehead and lateral headguard areas were tested. Peak headform acceleration was measured. Tests from a selection of drop heights and repeated tests on the same headguard were conducted. Headguard performance varied by test condition. For the 0.4 m rigid anvil tests, the best model headguard was the thickest producing an average peak headform acceleration over 5 tests of 48 g compared with 456 g for the worst model. The mean peak acceleration for the 0.4, 0.5 and 0.6 frontal and lateral rigid anvil impact tests was between 32% and 40% lower for the Top Ten boxing model compared with the Adidas boxing model. The headguard performance deterioration observed with repeat impact against the flat anvil was reduced for impacts against the glove section. The overall reduction in acceleration for the combination of glove and headguard in comparison to the headguard condition was in the range of 72-93% for 0.6 and 0.8 m drop tests. The impact tests show the benefits of performance testing in identifying differences between headguard models.


A meta-analysis of longitudinal partial correlations between school violence and mental health, school performance, and criminal or delinquent acts.
The daily challenges resulting from all types of school violence-such as physical aggression, bullying, peer victimization, and general threats-have the potential to affect, longitudinally, students' mental health, school performance, and involvement in criminal or delinquent acts. Across primary and secondary studies, however, variation in how and how much school violence relates to these outcomes, has persisted. The purpose of this systematic review and meta-analysis, therefore, was to clarify this uncertainty by synthesizing the longitudinal relations. We conducted exhaustive searching procedures, implemented rigorous screening and coding processes, and estimated an underused effect size, the partial correlation from multiple regression models, before estimating a random-effects meta-analysis using robust variance estimation. We meta-analyzed 114 independent studies, totaling 765 effect sizes across 95,618 individual participants. The results of the overall analyses found a statistically significant longitudinal relation between school violence, in any role, and the aggregated outcome variables (rp = .06). Given that this effect size inherently controls for multiple potential confounding covariates, we consider the relation's magnitude clinically meaningful. We end by discussing ways practitioners and researchers may use these analyses when implementing prevention programming and how the field of meta-analysis should more frequently utilize the partial correlation. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Age-related lymphocyte and neutrophil levels in children of hepatitis C-infected women.
Investigation of immunologic values in children vertically exposed to hepatitis C virus (HCV) infection could help explain the higher risk of infection in girls and indicate mechanisms of spontaneous viral clearance and possible long-term immunologic effects. Prospective study of children born to HCV-infected women. Lymphocyte and neutrophil measurements were age-standardized using the LMS method (this summarizes the changing age distribution of a variable). Associations between maternal and infant characteristics and lymphocyte and neutrophil z-scores were quantified using linear regression allowing for repeated measures. HCV-infected children, girls, and those born to HCV/human immunodeficiency virus (HIV)-coinfected women had significantly higher lymphocyte z-scores than HCV-uninfected children, boys, and children born to HCV-only-infected women, respectively. Peak absolute lymphocytes were significantly lower for infected children with evidence of viral clearance than for persistently infected children. Girls also had significantly higher neutrophil z-scores than boys but HCV-infected children had significantly lower neutrophil z-scores than uninfected children. The gender associations are in line with those observed among children born to HIV-infected women, suggesting general gender-based differences in response to infection. Age-related standards for uninfected children could be used to assess immune function in other pediatric diseases and these results suggest that gender-specific reference values should be used at least for the first 2 years of life.


The COLON study: Colorectal cancer: Longitudinal, Observational study on Nutritional and lifestyle factors that may influence colorectal tumour recurrence, survival and quality of life.
There is clear evidence that nutrition and lifestyle can modify colorectal cancer risk. However, it is not clear if those factors can affect colorectal cancer treatment, recurrence, survival and quality of life. This paper describes the background and design of the "COlorectal cancer: Longitudinal, Observational study on Nutritional and lifestyle factors that may influence colorectal tumour recurrence, survival and quality of life" - COLON - study. The main aim of this study is to assess associations of diet and other lifestyle factors, with colorectal cancer recurrence, survival and quality of life. We extensively investigate diet and lifestyle of colorectal cancer patients at diagnosis and during the following years; this design paper focusses on the initial exposures of interest: diet and dietary supplement use, body composition, nutrient status (e.g. vitamin D), and composition of the gut microbiota. The COLON study is a multi-centre prospective cohort study among at least 1,000 incident colorectal cancer patients recruited from 11 hospitals in the Netherlands. Patients with colorectal cancer are invited upon diagnosis. Upon recruitment, after 6 months, 2 years and 5 years, patients fill out food-frequency questionnaires; questionnaires about dietary supplement use, physical activity, weight, height, and quality of life; and donate blood samples. Diagnostic CT-scans are collected to assess cross-sectional areas of skeletal muscle, subcutaneous fat, visceral fat and intermuscular fat, and to assess muscle attenuation. Blood samples are biobanked to facilitate future analyse of biomarkers, nutrients, DNA etc. Analysis of serum 25-hydroxy vitamin D levels, and analysis of metabolomic profiles are scheduled. A subgroup of patients with colon cancer is asked to provide faecal samples before and at several time points after colon resection to study changes in gut microbiota during treatment. For all patients, information on vital status is retrieved by linkage with national registries. Information on clinical characteristics is gathered from linkage with the Netherlands Cancer Registry and with hospital databases. Hazards ratios will be calculated for dietary and lifestyle factors at diagnosis in relation to recurrence and survival. Repeated measures analyses will be performed to assess changes over time in dietary and other factors in relation to recurrence and survival.


Sex Difference in Draft-Legal Ultra-Distance Events - A Comparison between Ultra-Swimming and Ultra-Cycling.
Recent studies reported that the sex difference in performance in ultra-endurance sports such as swimming and cycling changed over the years. However, the aspect of drafting in draft-legal ultra-endurance races has not yet been investigated. This study investigates the sex difference in ultra-swimming and ultra-cycling draft-legal races where drafting - swimming or cycling behind other participants to save energy and have more power at the end of the race to overtake them, is allowed. The change in performance of the annual best and the annual three best in an ultra-endurance swimming race (16-km 'Faros Swim Marathon') over 38 years and in a 24-h ultra-cycling race ('World Cycling Race') over 13 years were compared and analysed with respect to sex difference. Furthermore, performances of the fastest female and male finishers ever were compared. In the swimming event, the sex difference of the annual best male and female decreased non-significantly (P = 0.262) from 5.3% (1976) to 1.0% (2013). The sex gap of speed in the annual three fastest swimmers decreased significantly (P = 0.043) from 5.9 ± 1.6% (1979) to 4.7 ± 3.1% (2013). In the cycling event, the difference in cycling speed between the annual best male and female decreased significantly (P = 0.026) from 33.31% (1999) to 10.89% (2011). The sex gap of speed in the annual three fastest decreased significantly (P = 0.001) from 32.9 ± 0.6% (1999) to 16.4 ± 5.9% (2011). The fastest male swimmer ever (swimming speed 5.3 km/h, race time: 03:01:55 h:min:s) was 1.5% faster than the fastest female swimmer (swimming speed 5.2 km/h, race time: 03:04:09 h:min:s). The three fastest male swimmers ever (mean 5.27 ± 0.13 km/h) were 4.4% faster than the three fastest female swimmers (mean 5.05 ± 0.20 km/h) (P < 0.05). In the cycling event, the best male ever (cycling speed 45.8 km/h) was 26.4% faster than the best female (cycling speed 36.1 km/h). The three fastest male cyclists ever (45.9 km/h) (mean 45.85 ± 0.05 km/h) were 32.1% faster (P < 0.05) than the three fastest female cyclists ever (34.7 km/h) (mean 34.70 ± 1.87 km/h). In summary, in draft-legal ultra-distance events such as swimming and cycling, the sex difference in the annual top and annual top three swimmers and cyclists decreased (i.e. non-linearly in swimmers and linearly in cyclists) over the years. The sex difference of the fastest athletes ever was smaller in swimming (1.5%) than in cycling (26.4%). This finding is different from reports about races where drafting was not possible or even prohibited and where the sex difference remained stable over years.


Diffusion tensor imaging and colored fractional anisotropy mapping of the ventralis intermedius nucleus of the thalamus.
The ventralis intermedius (VIM) nucleus of the thalamus is the primary surgical target for treatment of tremor. Most centers rely on indirect targeting based on atlas-defined coordinates rather than patient-specific anatomy, making intraoperative physiological mapping critical. Detailed identification of this target based on patient-specific anatomic features can help optimize the surgical treatment of tremor. To study colored fractional anisotropic images and diffusion tensor imaging (DTI) tractography to identify characteristic magnetic resonance appearances of the VIM nucleus. Four patients undergoing stereotactic surgery for essential tremor (ET) were retrospectively studied with analysis of magnetic resonance imaging-based colored fractional anisotropy (FA) images and fiber tractography. All were scanned with a 1.5-T magnetic resonance imaging unit, and all sequences were obtained before frame placement. Because the goal of this study was to identify the DTI characteristics of physiologically defined VIM nucleus, we selected and studied patients who had undergone DTI and had efficacious tremor control with intraoperative microlesioning effect and tremor reduction with less than 2.0-V stimulation. Analysis of color FA maps, which graphically illustrate fiber directionality, revealed consistent anatomic patterns. The region of the VIM nucleus can be seen as an intermediate region where there is a characteristic transition of color. Presumptive VIM nucleus interconnectivity with sensorimotor cortex and cerebellum was identified via the internal capsule and the superior cerebellar peduncle, respectively. FA maps could also be used to distinguish segments of gray matter, white matter, and gray-white matter boundaries. Analysis of DTI and FA maps on widely available 1.5-T magnetic resonance imaging yields clear identification of various structures key to neurosurgical targeting. Prospective evaluation of integrating DTI into neurosurgical planning may be warranted.


Residual dipolar couplings: are multiple independent alignments always possible?
RDCs for the 14 kDa protein hen egg-white lysozyme (HEWL) have been measured in eight different alignment media. The elongated shape and strongly positively charged surface of HEWL appear to limit the protein to four main alignment orientations. Furthermore, low levels of alignment and the protein's interaction with some alignment media increases the experimental error. Together with heterogeneity across the alignment media arising from constraints on temperature, pH and ionic strength for some alignment media, these data are suitable for structure refinement, but not the extraction of dynamic parameters. For an analysis of protein dynamics the data must be obtained with very low errors in at least three or five independent alignment media (depending on the method used) and so far, such data have only been reported for three small 6-8 kDa proteins with identical folds: ubiquitin, GB1 and GB3. Our results suggest that HEWL is likely to be representative of many other medium to large sized proteins commonly studied by solution NMR. Comparisons with over 60 high-resolution crystal structures of HEWL reveal that the highest resolution structures are not necessarily always the best models for the protein structure in solution.


Implementation of integrated stepped care for unhealthy alcohol use in HIV clinics.
Effective counseling and pharmacotherapy for unhealthy alcohol use are rarely provided in HIV treatment settings to patients. Our goal was to describe factors influencing implementation of a stepped care model to address unhealthy alcohol use in HIV clinics from the perspectives of social workers, psychologists and addiction psychiatrists. We conducted two focus groups with Social Workers (n = 4), Psychologists (n = 2), and Addiction Psychiatrists (n = 4) involved in an ongoing randomized controlled trial evaluating the effectiveness of integrated stepped care for unhealthy alcohol use in HIV-infected patients at five Veterans Health Administration (VA) HIV clinics. Data collection and analyses were guided by the Consolidated Framework for Implementation Research (CFIR) domains, with a focus on the three domains which we considered to be most relevant: intervention characteristics (i.e. motivational interviewing, pharmacotherapy), the inner setting (i.e. HIV clinics), and characteristics of individuals (i.e. the providers). A multidisciplinary team used directed content analysis to identify major themes. From the providers' perspective, the major implementation themes that emerged by CFIR domain included: (1) Intervention characteristics: providers valued tools and processes for facilitating patient motivation for treatment of unhealthy alcohol use given their perceived lack of motivation, but expressed a desire for greater flexibility; (2) Inner setting: treating unhealthy alcohol use in HIV clinics was perceived by providers to be consistent with VA priorities; and (3) Characteristics of individuals: there was high self-efficacy to conduct the intervention, an expressed need for more consistent utilization to maintain skills, and consideration of alternative models for delivering the components of the intervention. Use of the CFIR framework reveals that implementation of integrated stepped care for unhealthy alcohol use in HIV clinics is facilitated by tools to help providers enhance patient motivation or address unhealthy alcohol use among patients perceived to be unmotivated. Implementation may be facilitated by its consistency with organizational values and existing models of care and attention to optimizing provider self-efficacy and roles (i.e. approaches to treatment integration).


Primary care and health outcomes among older patients with diabetes.
The aim of this study was to measure the relationship between days spent waiting for primary care and health outcomes among patients diagnosed with diabetes, especially among the elderly population. Secondary data from VA administrative databases and Medicare claims. This is a retrospective observational study. Outcome variables include primary care utilization, mortality, heart attack, stroke, and ambulatory-care sensitive condition (ACSC) hospitalization. The main explanatory variable of interest is VA primary care wait time. Negative binomial models predict utilization and stacked logistic regression models predict the probability of experiencing each health outcome. Models are stratified by the presence of a selected health condition and age. Longer wait times were predicted to decrease utilization between 2 and 4 percent. There was no significant relationship between wait times and health outcomes for the overall sample. In stratified analyses, longer waits were associated with undesirable outcomes for those over age 70 with one of the selected health conditions or in certain narrower 5-year age groups, but the overall pattern of results does not indicate a systematic and significant effect. There was a modest effect of long wait times on primary care utilization but no robust effect of longer wait times on health outcomes. Waiting for care did not significantly compromise long-term health outcomes for veterans with diabetes.


Analysis of telomerase by the telomeric hemin/G-quadruplex-controlled aggregation of au nanoparticles in the presence of cysteine.
Telomeres are guanosine-rich nucleic-acid chains that fold, in the presence of K(+) ions and hemin, into the telomeric hemin/G-quadruplex structure, exhibiting horseradish peroxidase mimicking functions. The telomeric hemin/G-quadruplex structures catalyze the oxidation of thiols (e.g., l-cysteine) into disulfides (e.g., cystine). As l-cysteine stimulates the aggregation of Au nanoparticles (NPs), accompanied by absorbance changes from red (individual Au NPs) to blue (aggregated Au NPs), the process is implemented to quantitatively analyze the activity (content) of telomerase, a versatile biomarker for cancer cells. Telomerase extracted from 293T cancer cells catalyzes, in the presence of a dNTPs mixture and an appropriate primer probe, the telomerization process, leading to the generation of catalytic telomeric hemin/G-quadruplex chains that control the l-cysteine-mediated aggregation of Au NPs. The extent of aggregation is thus controlled by the concentration of telomerase. The method enabled the detection of telomerase with a detection limit of 27 cells/μL. The spectral changes accompanying the aggregation of Au NPs are further supported by transmission electron microscopy imaging.


Andexanet Alfa for the Reversal of Factor Xa Inhibitor Activity.
Bleeding is a complication of treatment with factor Xa inhibitors, but there are no specific agents for the reversal of the effects of these drugs. Andexanet is designed to reverse the anticoagulant effects of factor Xa inhibitors. Healthy older volunteers were given 5 mg of apixaban twice daily or 20 mg of rivaroxaban daily. For each factor Xa inhibitor, a two-part randomized placebo-controlled study was conducted to evaluate andexanet administered as a bolus or as a bolus plus a 2-hour infusion. The primary outcome was the mean percent change in anti-factor Xa activity, which is a measure of factor Xa inhibition by the anticoagulant. Among the apixaban-treated participants, anti-factor Xa activity was reduced by 94% among those who received an andexanet bolus (24 participants), as compared with 21% among those who received placebo (9 participants) (P<0.001), and unbound apixaban concentration was reduced by 9.3 ng per milliliter versus 1.9 ng per milliliter (P<0.001); thrombin generation was fully restored in 100% versus 11% of the participants (P<0.001) within 2 to 5 minutes. Among the rivaroxaban-treated participants, anti-factor Xa activity was reduced by 92% among those who received an andexanet bolus (27 participants), as compared with 18% among those who received placebo (14 participants) (P<0.001), and unbound rivaroxaban concentration was reduced by 23.4 ng per milliliter versus 4.2 ng per milliliter (P<0.001); thrombin generation was fully restored in 96% versus 7% of the participants (P<0.001). These effects were sustained when andexanet was administered as a bolus plus an infusion. In a subgroup of participants, transient increases in levels of d-dimer and prothrombin fragments 1 and 2 were observed, which resolved within 24 to 72 hours. No serious adverse or thrombotic events were reported. Andexanet reversed the anticoagulant activity of apixaban and rivaroxaban in older healthy participants within minutes after administration and for the duration of infusion, without evidence of clinical toxic effects. (Funded by Portola Pharmaceuticals and others; ANNEXA-A and ANNEXA-R ClinicalTrials.gov numbers, NCT02207725 and NCT02220725.).


Development and evolution of adaptive polyphenisms.
Phenotypic plasticity is the primitive character state for most if not all traits. Insofar as developmental and physiological processes obey the laws of chemistry and physics, they will be sensitive to such environmental variables as temperature, nutrient supply, ionic environment, and the availability of various macro- and micronutrients. Depending on the effect this phenotypic plasticity has on fitness, evolution may proceed to select either for mechanisms that buffer or canalize the phenotype against relevant environmental variation or for a modified plastic response in which some ranges of the phenotypic variation are adaptive to particular environments. Phenotypic plasticity can be continuous, in which case it is called a reaction norm, or discontinuous, in which case it is called a polyphenism. Although the morphological discontinuity of some polyphenisms is produced by discrete developmental switches, most polyphenisms are due to discontinuities in the environment that induce only portions of what is in reality a continuous reaction norm. In insect polyphenisms, the environmental variable that induces the alternative phenotype is a token stimulus that serves as a predictor of, but is not itself, the environment to which the polyphenism is an adaptation. In all cases studied so far, the environmental stimulus alters the endocrine mechanism of metamorphosis by altering either the pattern of hormone secretion or the pattern of hormone sensitivity in different tissues. Such changes in the patterns of endocrine interactions result in the execution of alternative developmental pathways. The spatial and temporal compartmentalization of endocrine interactions has produced a developmental mechanism that enables substantial localized changes in morphology that remain well integrated into the structure and function of the organism.


The practice and clinical implications of tablet splitting in international health.
Tablet splitting is frequently performed to facilitate correct dosing, but the practice and implications in low-income settings have rarely been discussed. We selected eight drugs, with narrow therapeutic indices or critical dosages, frequently divided in the Lao PDR (Laos). These were split, by common techniques used in Laos, by four nurses and four laypersons. The mean percentage deviation from the theoretical expected weight and weight loss of divided tablets/capsules were recorded. Five of eight study drugs failed, on splitting, to meet European Pharmacopoeia recommendations for tablet weight deviation from the expected weight of tablet/capsule halves with 10% deviating by more than 25%. There was a significant difference in splitting accuracy between nurses and laypersons (P = 0.027). Coated and unscored tablets were less accurately split than uncoated (P = 0.03 and 0.0019 for each half) and scored (0.0001 for both halves) tablets. These findings have potential clinical implications on treatment outcome and the development of antimicrobial resistance. Investment by drug companies in a wider range of dosage units, particularly for narrow therapeutic index and critical dosage medicines, is strongly recommended.


Antiplatelet therapy versus observation in low-risk essential thrombocythemia with a CALR mutation.
The role of antiplatelet therapy as primary prophylaxis of thrombosis in low-risk essential thrombocythemia has not been studied in randomized clinical trials. We assessed the benefit/risk of low-dose aspirin in 433 patients with low-risk essential thrombocythemia (271 with a CALR mutation, 162 with a JAK2(V617F) mutation) who were on antiplatelet therapy or observation only. After a follow up of 2215 person-years free from cytoreduction, 25 thrombotic and 17 bleeding episodes were recorded. In CALR-mutated patients, antiplatelet therapy did not affect the risk of thrombosis but was associated with a higher incidence of bleeding (12.9 versus 1.8 episodes per 1000 patient-years, P=0.03). In JAK2(V617F)-mutated patients, low-dose aspirin was associated with a reduced incidence of venous thrombosis with no effect on the risk of bleeding. Coexistence of JAK2(V617F)-mutation and cardiovascular risk factors increased the risk of thrombosis, even after adjusting for treatment with low-dose aspirin (incidence rate ratio: 9.8; 95% confidence interval: 2.3-42.3; P=0.02). Time free from cytoreduction was significantly shorter in CALR-mutated patients with essential thrombocythemia than in JAK2(V617F)-mutated ones (median time 5 years and 9.8 years, respectively; P=0.0002) and cytoreduction was usually necessary to control extreme thrombocytosis. In conclusion, in patients with low-risk, CALR-mutated essential thrombocythemia, low-dose aspirin does not reduce the risk of thrombosis and may increase the risk of bleeding.


Economic costs of the foot and mouth disease outbreak in the United Kingdom in 2001.
The authors present estimates of the economic costs to agriculture and industries affected by tourism of the outbreak of foot and mouth disease (FMD) in the United Kingdom (UK) in 2001. The losses to agriculture and the food chain amount to about Pound Sterling3.1 billion. The majority of the costs to agriculture have been met by the Government through compensation for slaughter and disposal as well as clean-up costs. Nonetheless, agricultural producers will have suffered losses, estimated at Pound Sterling355 million, which represents about 20% of the estimated total income from farming in 2001. Based on data from surveys of tourism, businesses directly affected by tourist expenditure are estimated to have lost a similar total amount (between Pound Sterling2.7 and Pound Sterling3.2 billion) as a result of reduced numbers of people visiting the countryside. The industries which supply agriculture, the food industries and tourist-related businesses will also have suffered losses. However, the overall costs to the UK economy are substantially less than the sum of these components, as much of the expenditure by tourists was not lost, but merely displaced to other sectors of the economy. Overall, the net effect of FMD is estimated to have reduced the gross domestic product in the UK by less than 0.2% in 2001.


The developmental expression of the heat-shock response in Xenopus laevis.
We have investigated the Xenopus heat-shock response of somatic cells, oocytes and embryos. Xenopus defolliculated oocytes displayed a highly variable response to heat shock depending on the culture medium. Intact follicles, however, respond to heat stress by synthesizing an invariant pattern of hsps. Although a subset of the hsp70/68 complex is expressed constitutively in the absence of heat shock in oocytes and embryos (hsc70), actual induction of hsps in response to stress does not occur until the blastula stage when transcription of the zygotic genome is first activated. By gastrulation, the hsps of somatic cells, including members of the hsp30/26 complex, were expressed coordinately in response to heat shock. We further show that Xenopus hsps have different solubilities perhaps reflecting their different subcellular locations. The 26,000-30,000 Mr complex (hsp30/26) was present almost exclusively in a detergent-insoluble fraction, as was 25-50% of the hsp70/68 complex and greater than 50% of hsp56, suggesting that these hsps may be associated with the cytoskeleton during a heat shock. In contrast, the other Xenopus hsps (hsp86, hsp75 and hsp61) were totally solubilized in a low-salt buffer.


Exposure to Bullying and General Psychopathology: A Prospective, Longitudinal Study.
Although there is mounting evidence that the experience of being bullied associates with both internalizing and externalizing symptoms, it is not known yet whether the identified associations are specific to these symptoms, or shared between them. The primary focus of this study is to assess the prospective associations of bullying exposure with both general and specific (i.e., internalizing, externalizing) factors of psychopathology. This study included data from 6,210 children participating in the Avon Longitudinal Study of Parents and Children (ALSPAC). Child bullying was measured by self-report at ages 8 and 10 years. Child psychopathology symptoms were assessed by parent-interview, using the Development and Well-being Assessment (DAWBA) at ages 7 and 13 years. Bullying exposure significantly associated with the general psychopathology factor in early adolescence. In particular, chronically victimized youth exposed to multiple forms of bullying (i.e., both overt and relational) showed higher levels of general psychopathology. Bullying exposure also associated with both internalizing and externalizing factors from the correlated-factors model. However, the effect estimates for these factors decreased considerably in size and dropped to insignificant for the internalizing factor after extracting the shared variance that belongs to the general factor of psychopathology. Using an integrative longitudinal model, we found that higher levels of general psychopathology at age 7 also associated with bullying exposure at age 8 which, in turn, associated with general psychopathology at age 13 through its two-year continuity. Findings suggest that exposure to bullying is a risk factor for a more general vulnerability to psychopathology.


Mental health, personality, and parental rearing styles of adolescents with Internet addiction disorder.
The objectives of this study were to compare the personality profiles of adolescent males with and without Internet addiction disorder (IAD), and to determine if IAD is associated with specific parental rearing behaviors. A total of 304 subjects (204 IAD positive and 100 IAD negative controls) completed three instruments: Symptom Checklist-90-revision (SCL-90-R), Eysenck Personality Questionnaire Revised (EPQ-R), and Egna Minnen av Barndoms Uppfostran--'My Memories of Upbringing' (EMBU). SCL-90-R profiles of adolescents with IAD revealed comparatively higher mean scores for all of the nine domains, and significantly higher scores for obsessive-compulsive, interpersonal sensitivity, depression, anxiety, hostility, and paranoid ideation; the mean global symptom index of adolescents with IAD was also significantly higher by approximately 10%. EPQ profiles of adolescents with IAD showed that Internet-dependent individuals tended to exhibit a significantly lower degree of extraversion and a significantly higher degree of psychoticism when compared with the control group. EMBU profiles revealed that adolescents with IAD generally rated both maternal and paternal rearing practices as lacking in emotional warmth, being over-involved, rejecting, and punitive (mothers only). The results of this study confirm that IAD often occurs concurrently with mental symptoms and personality traits such as introversion and psychoticism. Adolescents with IAD consistently rated parental rearing behaviors as being over-intrusive, punitive, and lacking in responsiveness. These findings suggest that the influences of parenting style and family function are important factors in the development of Internet dependency.


Engineering growth factors for regenerative medicine applications.
Growth factors are important morphogenetic proteins that instruct cell behavior and guide tissue repair and renewal. Although their therapeutic potential holds great promise in regenerative medicine applications, translation of growth factors into clinical treatments has been hindered by limitations including poor protein stability, low recombinant expression yield, and suboptimal efficacy. This review highlights current tools, technologies, and approaches to design integrated and effective growth factor-based therapies for regenerative medicine applications. The first section describes rational and combinatorial protein engineering approaches that have been utilized to improve growth factor stability, expression yield, biodistribution, and serum half-life, or alter their cell trafficking behavior or receptor binding affinity. The second section highlights elegant biomaterial-based systems, inspired by the natural extracellular matrix milieu, that have been developed for effective spatial and temporal delivery of growth factors to cell surface receptors. Although appearing distinct, these two approaches are highly complementary and involve principles of molecular design and engineering to be considered in parallel when developing optimal materials for clinical applications. Growth factors are promising therapeutic proteins that have the ability to modulate morphogenetic behaviors, including cell survival, proliferation, migration and differentiation. However, the translation of growth factors into clinical therapies has been hindered by properties such as poor protein stability, low recombinant expression yield, and non-physiological delivery, which lead to suboptimal efficacy and adverse side effects. To address these needs, researchers are employing clever molecular and material engineering and design strategies to both improve the intrinsic properties of growth factors and effectively control their delivery into tissue. This review highlights examples of interdisciplinary tools and technologies used to augment the therapeutic potential of growth factors for clinical applications in regenerative medicine.


Spatio-Genetic and phenotypic modelling elucidates resistance and re-sensitisation to treatment in heterogeneous melanoma.
Although novel targeted therapies have significantly improved the overall survival of patients with advanced melanoma, understanding and combatting drug resistance remains a major clinical challenge. Using partial differential equations, we describe the evolution of a cellular population through time, space, and phenotype dimensions, in the presence of various drug species. We then use this framework to explore models in which resistance is attained by either mutations (irreversible) or plasticity (reversible). Numerical results suggest that punctuated evolutionary assumptions are more consistent with results obtained from murine melanoma models than gradual evolution. Furthermore, in the context of an evolving tumour cell population, sequencing the treatment, for instance applying immunotherapy before BRAF inhibitors, can increase treatment effectiveness. However, drug strategies which showed success within a spatially homogeneous tumour environment were unsuccessful under heterogeneous conditions, suggesting that spatio-environmental heterogeneity may be the greatest challenge to tumour therapies. Plastic metabolic models are additionally capable of reproducing the characteristic resistant tumour volume curves and predicting re-sensitisation to secondary waves of treatment observed in patient derived xenograft (PDX) melanomas treated with MEK and BRAF inhibitors. Nevertheless, secondary relapse due to a pre-adapted subpopulation, remaining after the first wave of treatment, results in a more rapid development of resistance. Our model provides a framework through which tumour resistance can be understood and would suggest that carefully phased treatments may be able to overcome the development of long-term resistance in melanoma.


Generation of transgenic mice with mild and severe retinal neovascularisation.
To generate a mouse model for slow progressive retinal neovascularisation through vascular endothelial growth factor (VEGF) upregulation. Transgenic mice were generated via microinjection of a DNA construct containing the human VEGF165 (hVEGF) gene driven by a truncated mouse rhodopsin promoter. Mouse eyes were characterised clinically and histologically and ocular hVEGF levels assayed by ELISA. One transgenic line expressing low hVEGF levels showed mild clinical changes such as focal fluorescein leakage, microaneurysms, venous tortuosity, capillary non-perfusion and minor neovascularisation, which remained stable up to 3 months postnatal. Histologically, there were some disturbance and thinning of inner and outer nuclear layers, with occasional focal areas of neovascularisation. By contrast, three other lines expressing high hVEGF levels presented with concomitantly severe phenotypes. In addition to the above, clinical features included extensive neovascularisation, haemorrhage, and retinal detachment; histologically, focal to extensive areas of neovascularisation associated with retinal folds, cell loss in the inner and outer nuclear layers, and partial retinal detachment were common. The authors generated four hVEGF overexpressing transgenic mouse lines with phenotypes ranging from mild to severe neovascularisation. These models are a valuable research tool to study excess VEGF related molecular and cellular changes and provide additional opportunities to test anti-angiogenic therapies.


Genetic transformation of Veillonella parvula.
Veillonellae are one of the most prevalent and predominant microorganisms in both the supra- and subgingival plaques of the human oral cavity. Veillonellae's mutualistic relationships with the early, middle, and late colonizers of the oral cavity make them an important component of oral biofilm ecology. Unlike other ubiquitous early colonizers in the oral cavity, surprisingly little is known about Veillonella biology due to our lack of ability to genetically transform this group of bacteria. The objective of this study was to test the transformability of veillonellae. Using Veillonella parvula strain PK1910, we first obtained spontaneous mutations conferring streptomycin resistance. These mutations all carry a K43N substitution in the RpsL protein. Using the mutated rpsL gene as a selection marker, a variety of conditions were tested and optimized for electroporation. With the optimized protocol, we were able to introduce the first targeted mutation into the chromosome of V. parvula PK1910. Although more studies are needed to develop a robust genetic manipulation system in veillonellae, our results demonstrated, for the first time, that V. parvula is transformable, at least for strain PK1910.


Dopamine D2 receptors in addiction-like reward dysfunction and compulsive eating in obese rats.
We found that development of obesity was coupled with emergence of a progressively worsening deficit in neural reward responses. Similar changes in reward homeostasis induced by cocaine or heroin are considered to be crucial in triggering the transition from casual to compulsive drug-taking. Accordingly, we detected compulsive-like feeding behavior in obese but not lean rats, measured as palatable food consumption that was resistant to disruption by an aversive conditioned stimulus. Striatal dopamine D2 receptors (D2Rs) were downregulated in obese rats, as has been reported in humans addicted to drugs. Moreover, lentivirus-mediated knockdown of striatal D2Rs rapidly accelerated the development of addiction-like reward deficits and the onset of compulsive-like food seeking in rats with extended access to palatable high-fat food. These data demonstrate that overconsumption of palatable food triggers addiction-like neuroadaptive responses in brain reward circuits and drives the development of compulsive eating. Common hedonic mechanisms may therefore underlie obesity and drug addiction.


Temperature effects on dielectric liquid lenses.
The thermal stability of dielectric liquid lenses is studied by measuring the focal length at different temperatures. Two types of liquids lenses are investigated: Type-I (SL-5267/glycerol) and Type-II (glycerol/ BK7 matching liquid). A threshold-like behavior is found. Below the threshold temperature, the focal length is temperature insensitive. Above the threshold, the focal length changes exponentially with the temperature. Both refractive index and surface profile are responsible for the focal length change, although the former decreases linearly with the temperature. The threshold temperature of Type-I and Type-II liquid lens are 60°C and 40°C, respectively. Type-I lens shows a good temperature stability in a wide range. Moreover, the lens can recover to its original state even though it is operated at a high temperature.


Gastro-Protective and Anti-Oxidant Potential of Althaea officinalis and Solanum nigrum on Pyloric Ligation/Indomethacin-Induced Ulceration in Rats.
Recently, an alternative disease treatment approach is the research of medicaments from traditional medicine. Plants with anti-oxidant capabilities are used as herbal treatments for ulcer diseases. Medicinal/herbal extracts containing phytoconstituents have significant anti-ulcer activities in in vivo experiments on animal models, compared to reference drugs. The current study aims to inspect gastro-protective as well as in vitro and in vivo anti-oxidant potential of Althaea officinalis and Solanum nigrum extracts on pyloric-ligation/indomethacin-induced gastric-ulceration in rats. Rats were divided into six groups: normal control, gastric ulcer control, two standard pretreatment groups receiving omeprazole and misoprostol, and two test pretreatment groups receiving Althaea officinalis and Solanum nigrum. Pretreatments were administrated orally for 14 days. On the 15th day, animals, excluding the normal control group, were exposed to pyloric-ligation followed by indomethacin injection. After four hours, the rat's stomachs were removed and gastric juice and blood samples were collected. Pyloric-ligation/indomethacin administration caused considerable elevation in ulcer number, ulcer index, acid and pepsin productivity, aggressive factors, and gastric mucosal lipid-peroxide contents. Moreover, reduction in titratable acidity, gastric mucosal nitric-oxide, anti-oxidant contents, and protective factors accompanied gastric-ulceration. Additionally, elevation in pro-inflammatory cytokines content and reduction in cystathionine-β-synthase and heme-oxygenase-1 expression was witnessed. Omeprazole, misoprostol, Althaea officinalis, and Solanum nigrum pretreatments fixed blood and tissue biomarkers, thereby protecting them from pyloric-ligation/indomethacin-induced gastric-ulceration in rats, which is hopeful for clinical examinations.


The ethics of the Texas death penalty and its impact on a prolonged appeals process.
Society remains sharply divided as to the deterrent value of capital punishment. Following the reintroduction of the death penalty in the United States, Texas law mandates the affirmative predictability of future dangerousness beyond a reasonable doubt before a jury can impose the ultimate penalty for capital murder. The validity of prediction of dangerousness has been challenged in three Texas landmark cases before the U.S. Supreme Court. The case of Karla Faye Tucker highlights the moral controversy that occurs when execution follows an appeals process stretching over more than a decade, during which time personality growth and the effects of prison rehabilitation may have eliminated or curbed criminal tendencies.


Macrophage heterogeneity in tissues: phenotypic diversity and functions.
During development and throughout adult life, macrophages derived from hematopoietic progenitors are seeded throughout the body, initially in the absence of inflammatory and infectious stimuli as tissue-resident cells, with enhanced recruitment, activation, and local proliferation following injury and pathologic insults. We have learned a great deal about macrophage properties ex vivo and in cell culture, but their phenotypic heterogeneity within different tissue microenvironments remains poorly characterized, although it contributes significantly to maintaining local and systemic homeostasis, pathogenesis, and possible treatment. In this review, we summarize the nature, functions, and interactions of tissue macrophage populations within their microenvironment and suggest questions for further investigation.


DEOD: uncovering dominant effects of cancer-driver genes based on a partial covariance selection method.
The generation of a large volume of cancer genomes has allowed us to identify disease-related alterations more accurately, which is expected to enhance our understanding regarding the mechanism of cancer development. With genomic alterations detected, one challenge is to pinpoint cancer-driver genes that cause functional abnormalities. Here, we propose a method for uncovering the dominant effects of cancer-driver genes (DEOD) based on a partial covariance selection approach. Inspired by a convex optimization technique, it estimates the dominant effects of candidate cancer-driver genes on the expression level changes of their target genes. It constructs a gene network as a directed-weighted graph by integrating DNA copy numbers, single nucleotide mutations and gene expressions from matched tumor samples, and estimates partial covariances between driver genes and their target genes. Then, a scoring function to measure the cancer-driver score for each gene is applied. To test the performance of DEOD, a novel scheme is designed for simulating conditional multivariate normal variables (targets and free genes) given a group of variables (driver genes). When we applied the DEOD method to both the simulated data and breast cancer data, DEOD successfully uncovered driver variables in the simulation data, and identified well-known oncogenes in breast cancer. In addition, two highly ranked genes by DEOD were related to survival time. The copy number amplifications of MYC (8q24.21) and TRPS1 (8q23.3) were closely related to the survival time with P-values = 0.00246 and 0.00092, respectively. The results demonstrate that DEOD can efficiently uncover cancer-driver genes.


Development and validation of a 36-gene sequencing assay for hereditary cancer risk assessment.
The past two decades have brought many important advances in our understanding of the hereditary susceptibility to cancer. Numerous studies have provided convincing evidence that identification of germline mutations associated with hereditary cancer syndromes can lead to reductions in morbidity and mortality through targeted risk management options. Additionally, advances in gene sequencing technology now permit the development of multigene hereditary cancer testing panels. Here, we describe the 2016 revision of the Counsyl Inherited Cancer Screen for detecting single-nucleotide variants (SNVs), short insertions and deletions (indels), and copy number variants (CNVs) in 36 genes associated with an elevated risk for breast, ovarian, colorectal, gastric, endometrial, pancreatic, thyroid, prostate, melanoma, and neuroendocrine cancers. To determine test accuracy and reproducibility, we performed a rigorous analytical validation across 341 samples, including 118 cell lines and 223 patient samples. The screen achieved 100% test sensitivity across different mutation types, with high specificity and 100% concordance with conventional Sanger sequencing and multiplex ligation-dependent probe amplification (MLPA). We also demonstrated the screen's high intra-run and inter-run reproducibility and robust performance on blood and saliva specimens. Furthermore, we showed that pathogenic Alu element insertions can be accurately detected by our test. Overall, the validation in our clinical laboratory demonstrated the analytical performance required for collecting and reporting genetic information related to risk of developing hereditary cancers.


Reconsidering the Use of Autoregressive Latent Trajectory (ALT) Models.
The simultaneous estimation of autoregressive (simplex) structures and latent trajectories, so called ALT (autoregressive latent trajectory) models, is becoming an increasingly popular approach to the analysis of change. Although historically autoregressive (AR) and latent growth curve (LGC) models have been developed quite independently from each other, the underlying pattern of change is often highly similar. In this article it is shown that their integration rests on the strong assumption that neither the AR part nor the LGC part contains any misspecification. In practice, however, this assumption is often violated due to nonlinearity in the LGC part. As a consequence, the autoregressive (simplex) process incorrectly accounts for part of this nonlinearity, thus rendering any substantive interpretation of parameter estimates virtually impossible. Accordingly, researchers are advised to exercise extreme caution when using ALT models in practice. All arguments are illustrated by empirical data on skill acquisition, and a simulation study is provided to investigate the conditions and consequences of mistaking nonlinear growth curve patterns as autoregressive processes.


Cost analysis of an integrated disease surveillance and response system: case of Burkina Faso, Eritrea, and Mali.
Communicable diseases are the leading causes of illness, deaths, and disability in sub-Saharan Africa. To address these threats, countries within the World Health Organization (WHO) African region adopted a regional strategy called Integrated Disease Surveillance and Response (IDSR). This strategy calls for streamlining resources, tools, and approaches to better detect and respond to the region's priority communicable disease. The purpose of this study was to analyze the incremental costs of establishing and subsequently operating activities for detection and response to the priority diseases under the IDSR. We collected cost data for IDSR activities at central, regional, district, and primary health care center levels from Burkina Faso, Eritrea, and Mali, countries where IDSR is being fully implemented. These cost data included personnel, transportation items, office consumable goods, media campaigns, laboratory and response materials and supplies, and annual depreciation of buildings, equipment, and vehicles. Over the period studied (2002-2005), the average cost to implement the IDSR program in Eritrea was $0.16 per capita, $0.04 in Burkina Faso and $0.02 in Mali. In each country, the mean annual cost of IDSR was dependent on the health structure level, ranging from $35,899 to $69,920 at the region level, $10,790 to $13,941 at the district level, and $1,181 to $1,240 at the primary health care center level. The proportions spent on each IDSR activity varied due to demand for special items (e.g., equipment, supplies, drugs and vaccines), service availability, distance, and the epidemiological profile of the country. This study demonstrates that the IDSR strategy can be considered a low cost public health system although the benefits have yet to be quantified. These data can also be used in future studies of the cost-effectiveness of IDSR.


FRET enhancement close to gold nanoparticles positioned in DNA origami constructs.
Here we investigate the energy transfer rates of a Förster resonance energy transfer (FRET) pair positioned in close proximity to a 5 nm gold nanoparticle (AuNP) on a DNA origami construct. We study the distance dependence of the FRET rate by varying the location of the donor molecule, D, relative to the AuNP while maintaining a fixed location of the acceptor molecule, A. The presence of the AuNP induces an alteration in the spontaneous emission of the donor (including radiative and non-radiative rates) which is strongly dependent on the distance between the donor and AuNP surface. Simultaneously, the energy transfer rates are enhanced at shorter D-A (and D-AuNP) distances. Overall, in addition to the direct influence of the acceptor and AuNP on the donor decay there is also a significant increase in decay rate not explained by the sum of the two interactions. This leads to enhanced energy transfer between donor and acceptor in the presence of a 5 nm AuNP. We also demonstrate that the transfer rate in the three "particle" geometry (D + A + AuNP) depends approximately linearly on the transfer rate in the donor-AuNP system, suggesting the possibility to control FRET process with electric field induced by 5 nm AuNPs close to the donor fluorophore. It is concluded that DNA origami is a very versatile platform for studying interactions between molecules and plasmonic nanoparticles in general and FRET enhancement in particular.


Fundamental limits to position determination by concentration gradients.
Position determination in biological systems is often achieved through protein concentration gradients. Measuring the local concentration of such a protein with a spatially varying distribution allows the measurement of position within the system. For these systems to work effectively, position determination must be robust to noise. Here, we calculate fundamental limits to the precision of position determination by concentration gradients due to unavoidable biochemical noise perturbing the gradients. We focus on gradient proteins with first-order reaction kinetics. Systems of this type have been experimentally characterised in both developmental and cell biology settings. For a single gradient we show that, through time-averaging, great precision potentially can be achieved even with very low protein copy numbers. As a second example, we investigate the ability of a system with oppositely directed gradients to find its centre. With this mechanism, positional precision close to the centre improves more slowly with increasing averaging time, and so longer averaging times or higher copy numbers are required for high precision. For both single and double gradients, we demonstrate the existence of optimal length scales for the gradients for which precision is maximized, as well as analyze how precision depends on the size of the concentration-measuring apparatus. These results provide fundamental constraints on the positional precision supplied by concentration gradients in various contexts, including both in developmental biology and also within a single cell.


Antitumor effects of a natural anthracycline analog (Aloin) involve altered activity of antioxidant enzymes in HeLaS3 cells.
The antiproliferative and cytotoxic potential of the natural anthracycline aloin from Aloe vera was tested on human uterine carcinoma HeLaS3 cells. Aloin showed a pronounced antiproliferative effect at physiological concentration (IC50 = 97 microM), caused cell cycle arrest in the S phase and markedly increased HeLaS3 cell apoptosis (to 24%). In the concentration range of 20-100 microM, its action was accompanied by remarkable changes in the activity of almost all antioxidant enzymes: MnSOD activity was increased many fold, while CuZnSOD and iNOS activities were inhibited. Moreover, inhibition of CuZnSOD was shown to occur by direct aloin interaction with the enzyme. As catalase activity was not changed, it is suggested that such conditions were responsible for antiproliferative and cytotoxic effects owing to accumulation of H2O2. Aloin alone was a more potent proapoptotic agent than a 2 Gy fractional dose of ionizing radiation or a combination of the two. Compared to other currently used therapeutics, aloin, due to its less undesirable side effects and antimetastatic potential, may prove to be the agent of choice on which clinical protocols for the treatment of human cervical carcinoma should rely in future.


Delayed convergence between brain network structure and function in rolandic epilepsy.
Rolandic epilepsy (RE) manifests during a critical phase of brain development, and has been associated with language impairments. Concordant abnormalities in structural and functional connectivity (SC and FC) have been described before. As SC and FC are under mutual influence, the current study investigates abnormalities in the SC-FC synergy in RE. Twenty-two children with RE (age, mean ± SD: 11.3 ± 2.0 y) and 22 healthy controls (age 10.5 ± 1.6 y) underwent structural, diffusion weighted, and resting-state functional magnetic resonance imaging (MRI) at 3T. The probabilistic anatomical landmarks atlas was used to parcellate the (sub)cortical gray matter. Constrained spherical deconvolution tractography and correlation of time series were used to assess SC and FC, respectively. The SC-FC correlation was assessed as a function of age for the non-zero structural connections over a range of sparsity values (0.01-0.75). A modularity analysis was performed on the mean SC network of the controls to localize potential global effects to subnetworks. SC and FC were also assessed separately using graph analysis. The SC-FC correlation was significantly reduced in children with RE compared to healthy controls, especially for the youngest participants. This effect was most pronounced in a left and a right centro-temporal network, as well as in a medial parietal network. Graph analysis revealed no prominent abnormalities in SC or FC network organization. Since SC and FC converge during normal maturation, our finding of reduced SC-FC correlation illustrates impaired synergy between brain structure and function. More specifically, since this effect was most pronounced in the youngest participants, RE may represent a developmental disorder of delayed brain network maturation. The observed effects seem especially attributable to medial parietal connections, which forms an intermediate between bilateral centro-temporal modules of epileptiform activity, and bear relevance for language function.


WNT2 regulates DNA synthesis in mouse granulosa cells through beta-catenin.
WNTs are secreted extracellular signaling molecules that transduce their signals by binding to G protein-coupled receptors of the frizzled (FZD) family. They control diverse developmental processes, such as cell fate specification, cell proliferation, cell differentiation, and apoptosis. Although WNT signaling has been shown to be essential for development of the ovary, its mechanistic role in folliculogenesis within the adult ovary has not been studied extensively. Therefore, the objective of this study was to investigate the regulation and function of WNT2 signaling in mouse granulosa cells. Immunostaining identified WNT2 as being expressed in granulosa cells throughout folliculogenesis, but with varying signal strength: in sequential sections, WNT2 immunoreactivity was strongest in healthy antral follicles but weak in atretic follicles. Knockdown of WNT2 expression using transfected short interfering RNA decreased DNA synthesis in granulosa cells, whereas WNT2 overexpression using a recombinant viral vector enhanced it. WNT2 knockdown led to accumulation of glycogen synthase kinase-3beta (GSK3B) in the cytoplasm but reduced the expression of beta-catenin. Conversely, WNT2 overexpression reduced the expression of GSK3B in the cytoplasm and induced beta-catenin translocation from the membrane into the nucleus. Beta-catenin knockdown also inhibited DNA synthesis in granulosa cells and neutralized the effect of WNT2 overexpression. WNT2/beta-catenin signaling had a slight effect on the apoptosis of granulosa cells. Taken together, the data indicate that WNT2 regulates beta-catenin localization in granulosa cells, and WNT2/beta-catenin signaling contributes to regulating their proliferation.


Relapse rates and enhancing lesions in a phase II trial of natalizumab in multiple sclerosis.
Natalizumab, a humanized monoclonal IgG4 antibody, is an alpha4-integrin antagonist, which inhibits the migration of inflammatory cells into the central nervous system, a key pathogenic mechanism in multiple sclerosis (MS). In a six month, phase II clinical trial of patients with relapsing MS, natalizumab significantly reduced the formation of new gadolinium-enhanced (Gd+) lesions and the number of clinical relapses. To investigate the relationship of historical relapse rate and new Gd + lesion number with subsequent MS disease activity and natalizumab treatment in the phase II study. Patients who participated in the phase II study were stratified into subgroups according to: (i) the number of relapses in the two years prior to entry into the study: 2 relapses (n = 108), 3 relapses (n =57), and >3 relapses (n =48); (ii) the number of new Gd + lesions at baseline (Month 0): 0 (n = 129), 1-2 (n =50), and >2 (n =33). Relapses and new Gd + lesions during the treatment phase of the trial were determined and compared for each subgroup. Both the prestudy relapse rate and number of new Gd + lesions at baseline were related to the subsequent risk of a relapse; baseline number of Gd + lesions was related to the likelihood of subsequent new Gd + lesion formation. There was a lower proportion of subjects with an on-study relapse and fewer new Gd + lesions in all natalizumab-treated subgroups when compared with their placebo counterpart; the difference was most apparent in the subgroups of patients with >3 relapses in the two years prior to study entry and >2 new Gd -- lesions at Month 0. There was a lower proportion of subjects with an on-study relapse in natalizumab-treated patients, particularly in those with a more active disease at study entry. Larger ongoing phase III studies will allow more definitive investigation of these preliminary subgroup findings.


Metabarcoding a diverse arthropod mock community.
Although DNA metabarcoding is an attractive approach for monitoring biodiversity, it is often difficult to detect all the species present in a bulk sample. In particular, sequence recovery for a given species depends on its biomass and mitome copy number as well as the primer set employed for PCR. To examine these variables, we constructed a mock community of terrestrial arthropods comprised of 374 species. We used this community to examine how species recovery was impacted when amplicon pools were constructed in four ways. The first two protocols involved the construction of bulk DNA extracts from different body segments (Bulk Abdomen, Bulk Leg). The other protocols involved the production of DNA extracts from single legs which were then merged prior to PCR (Composite Leg) or PCR-amplified separately (Single Leg) and then pooled. The amplicons generated by these four treatments were then sequenced on three platforms (Illumina MiSeq, Ion Torrent PGM and Ion Torrent S5). The choice of sequencing platform did not substantially influence species recovery, although the Miseq delivered the highest sequence quality. As expected, species recovery was most efficient from the Single Leg treatment because amplicon abundance varied little among taxa. Among the three treatments where PCR occurred after pooling, the Bulk Abdomen treatment produced a more uniform read abundance than the Bulk Leg or Composite Leg treatment. Primer choice also influenced species recovery and evenness. Our results reveal how variation in protocols can have substantial impacts on perceived diversity unless sequencing coverage is sufficient to reach an asymptote.


Improving mental health in health care practitioners: randomized controlled trial of a gratitude intervention.
Chronic occupational stress is common among health care practitioners, with potential impacts on personal mental health and staff turnover. This study investigated whether directing practitioners' attention to thankful events in work could reduce stress and depressive symptoms. A double-blind randomized controlled trial was conducted in 5 public hospitals with follow-up to 3 months posttreatment. One hundred two practitioners were randomly assigned into 3 conditions: gratitude, hassle, and nil-treatment. Those with scheduled long leaves were excluded. Participants in the gratitude and hassle group wrote work-related gratitude and hassle diaries respectively twice a week for 4 consecutive weeks. A no-diary group served as control. Depressive symptoms (primary outcome) and perceived stress (secondary outcome) were collected at baseline, posttreatment, and 3-month follow-up. Intent-to-treat analyses were performed with mixed-effects regression. Significant Treatment × Time interaction effects were found for the gratitude intervention, whether it was compared with control or hassle; the general pattern was a decline in stress and depressive symptoms over time, but the rate of decline became less pronounced as time progressed. Hassle and control were basically indistinct from each other. Relative to control, the gratitude group reported lower depressive symptoms (-1.50 points; 95% CI [-2.98, -0.01]; d = -0.49) and perceived stress (-2.65 points; 95% CI [-4.00, -1.30]; d = -0.95) at follow-up. RESULTS for the comparison between gratitude and hassle were similar. Taking stock of thankful events is an effective approach to reduce stress and depressive symptoms among health care practitioners.


Bisphosphonamidate clodronate prodrug exhibits selective cytotoxic activity against melanoma cell lines.
Bisphosphonates are used clinically to treat disorders of calcium metabolism and malignant bone disease and are known to inhibit cancer cell growth, adhesion, and invasion. However, clinical use of these agents for the treatment of extraskeletal disease is limited because of low cell permeability. We recently described a bisphosphonamidate prodrug strategy for efficient intracellular release of bisphosphonates, including clodronate (CLO), in non-small cell lung cancer cells. To evaluate anticancer activity of this prodrug class across many cancer cell types, the bisphosphonamidate clodronate prodrug (CLO prodrug) was screened against the NCI-60 cell line panel, and was found to exhibit selectivity toward melanoma cell lines. Here, we confirm efficient cellular uptake and intracellular activation of this prodrug class in melanoma cells. We further demonstrate inhibition of melanoma cell proliferation, induction of apoptosis, and an antitumor effect of CLO prodrug in a xenograft model. These data suggest a novel therapeutic application for the CLO prodrug and potential to selectively target melanoma cells.


Viral fusion: how Flu induces dimples on liposomes.
Membrane fusion--a basic cellular process enabling crucial cellular functions such as membrane trafficking--is mechanistically only partially understood. Most of the existing knowledge has come from studying host-cell entry of viruses with lipid bilayer envelopes, which proceeds through fusion of viral and host-cell membranes.Crystal structures for a number of viral fusion proteins facilitating this process have contributed substantially to our understanding. One of the next challenges is to merge these high-resolution structures of soluble parts of fusion effectors with native structural information on the proteins in the course of their functional interactions with the target membranes. In this issue of The EMBO Journal,Lee (2010) presents an elegant example of such a study,probing the early interaction of the influenza virus with liposomes by cryo-electron tomography (cryoET).


Chemically addressed switching measurements in graphene electrode memristive devices using in situ XPS.
The memristor has been the topic of heated research activity since the Pt/TiO2/TiOx/Pt structure was reported by Hewlett-Packard Labs. The characteristics of memristors such as the pinched hysteresis loops and time and input signal-dependent memristance are due to the drift of positively charged oxygen vacancies in the TiOx layer. While different modes of switching behaviour have been characterized, observing the switching as it happens with in situ measurements using X-ray photoelectron spectroscopy (XPS) can allow a better understanding of the device operation. The setup used in this work enables the application of voltage signals of different frequencies and amplitudes and observing the hysteresis seen in the I-V plane through chemical addressing. The measurements were conducted on Pt/TiO2/TiOx/graphene structures. The single layer graphene, utilized as a top electrode, effectively acts as a transparent electrode in that the layer beneath it can be observed to a depth of ∼10 nm in XPS. This allows for the observation of the changes in the binding energies of C 1s, Ti 2p and O 1s. By comparing the variation in the binding energy of Ti 2p to that of C 1s, and observing how the variation changes for different excitation signals (at different frequency and amplitudes), it is possible to inspect the effect of the oxygen vacancy drift. We employed a variety of input signals with varying frequency and amplitudes in order to test the memristive devices thoroughly: sine wave, triangular wave and DC bias. Graphene has been attracting attention due to its intriguing optoelectronic properties. This study utilizes graphene as a transparent top electrode for in situ measurements in XPS to observe chemically-addressed memristive hysteresis while an excitation signal is being applied to the device.


Evidence for D0-D(0) mixing using the CDF II detector.
We measure the time dependence of the ratio of decay rates for the rare decay D{0}-->K{+}pi{-} to the Cabibbo-favored decay D{0}-->K{-}pi;{+}. A signal of 12.7x10;{3} D{0}-->K{+}pi{-} decays was obtained using the Collider Detector at Fermilab II detector at the Fermilab Tevatron with an integrated luminosity of 1.5 fb;{-1}. We measure the D0-D[over ]{0} mixing parameters (R_{D},y{'},x{'2}), and find that the data are inconsistent with the no-mixing hypothesis with a probability equivalent to 3.8 Gaussian standard deviations.


Simulating Cross-Contamination of Cooked Pork with Salmonella enterica from Raw Pork through Home Kitchen Preparation in Vietnam.
Pork is the most commonly consumed meat in Vietnam, and Salmonella enterica is a common contaminant. This study aimed to assess potential S. enterica cross-contamination between raw and cooked pork in Vietnamese households. Different scenarios for cross-contamination were constructed based on a household survey of pork handling practices (416 households). Overall, 71% of people used the same knife and cutting board for both raw and cooked pork; however, all washed their hands and utensils between handling raw and cooked pork. The different scenarios were experimentally tested. First, S. enterica was inoculated on raw pork and surfaces (hands, knives and cutting boards); next, water used for washing and pork were sampled to identify the presence and concentration of S. enterica during different scenarios of food preparation. Bootstrapping techniques were applied to simulate transfer rates of S. enterica cross-contamination. No cross-contamination to cooked pork was observed in the scenario of using the same hands with new cutting boards and knives. The probability of re-contamination in the scenarios involving re-using the cutting board after washing was significantly higher compared to the scenarios which used a new cutting board. Stochastic simulation found a high risk of cross-contamination from raw to cooked pork when the same hands, knives and cutting boards were used for handling raw and cooked pork (78%); when the same cutting board but a different knife was used, cross-contamination was still high (67%). Cross-contamination between was not seen when different cutting boards and knives were used for cutting raw and cooked pork. This study provided an insight into cross-contamination of S. enterica, given common food handling practices in Vietnamese households and can be used for risk assessment of pork consumption.


Identical accumulation and immobilization of sulfated and nonsulfated Nod factors in host and nonhost root hair cell walls.
Nod factors are signaling molecules secreted by Rhizobium bacteria. These lipo-chitooligosaccharides (LCOs) are required for symbiosis with legumes and can elicit specific responses at subnanomolar concentrations on a compatible host. How plants perceive LCOs is unclear. In this study, using fluorescent Nod factor analogs, we investigated whether sulfated and nonsulfated Nod factors were bound and perceived differently by Medicago truncatula and Vicia sativa root hairs. The bioactivity of three novel sulfated fluorescent LCOs was tested in a root hair deformation assay on M. truncatula, showing bioactivity down to 0.1 to 1 nM. Fluorescence microscopy of plasmolyzed M. truncatula root hairs shows that sulfated fluorescent Nod factors accumulate in the cell wall of root hairs, whereas they are absent from the plasma membrane when applied at 10 nM. When the fluorescent Nod factor distribution in medium surrounding a root was studied, a sharp decrease in fluorescence close to the root hairs was observed, visualizing the remarkable capacity of root hairs to absorb Nod factors from the medium. Fluorescence correlation microscopy was used to study in detail the mobilities of sulfated and nonsulfated fluorescent Nod factors which are biologically active on M. truncatula and V. sativa, respectively. Remarkably, no difference between sulfated and nonsulfated Nod factors was observed: both hardly diffuse and strongly accumulate in root hair cell walls of both M. truncatula and V. sativa. The implications for the mode of Nod factor perception are discussed.


Association among agro-industrial traits and simultaneous selection in sweet sorghum.
Sweet sorghum has emerged as an alternative crop for ethanol yield. The breeding of this crop is performed to obtain cultivars with high ethanol yield, which necessarily requires associating favorable phenotypes for multiple traits. Therefore, the aims of this study were to investigate the association between agro-industrial traits related to ethanol yield and identify the promising genotypes considering multiple traits in sweet sorghum. For this purpose, we evaluated 45 genotypes using a 9 x 5 alpha-lattice experimental design with three replications. The traits measured were flowering time, plant height, tons of stalk per hectare, total soluble solids, tons of brix per hectare, juice extraction, total recoverable sugars, and ethanol yield. Analyses were performed after the recovery of inter-block information. The interrelation of the traits was described by genotype-by-trait biplot. For simultaneous selection, the Modified Mulamba and Mock index was used. For almost all of the agro-industrial traits, except for juice extraction, selective accuracy was above 70%. There were significant differences among genotypes for all the traits. The genotype-by-trait biplot evidenced a positive association between most of the traits related to ethanol yield, except for juice extraction, indicating the possibility of indirect selection to obtain more productive genotypes. Some genotypes proved to be promising based on the selection index, as they accumulated phenotypes favorable for the traits of interest.


Reliability-guided digital image correlation for image deformation measurement.
A universally applicable reliability-guided digital image correlation (DIC) method is proposed for reliable image deformation measurement. The zero-mean normalized cross correlation (ZNCC) coefficient is used to identify the reliability of the point computed. The correlation calculation begins with a seed point and is then guided by the ZNCC coefficient. That means the neighbors of the point with the highest ZNCC coefficient in a queue for computed points will be processed first. Thus the calculation path is always along the most reliable direction, and possible error propagation of the conventional DIC method can be avoided. The proposed novel DIC method is universally applicable to the images with shadows, discontinuous areas, and deformation discontinuity. Two image pairs were used to evaluate the performance of the proposed technique, and the successful results clearly demonstrate its robustness and effectiveness.


Greater daily leisure time physical activity is associated with lower chronic disease risk in adults with spinal cord injury.
The objective of this study was to examine the relationship between leisure time physical activity (LTPA) and common risk factors for cardiovascular disease (CVD) and type 2 diabetes in community-dwelling adults with chronic spinal cord injury (SCI). LTPA was measured using the Physical Activity Recall Assessment for People with SCI in 76 men and women with chronic (> or =1 year) paraplegia or tetraplegia, living in or near Hamilton, Ontario. Body mass index (BMI), waist circumference, body composition (fat mass (FM) and fat-free mass (FFM)), blood pressure, and biochemical data were collected. Thirty-seven percent (n = 28 participants) were inactive, reporting no LTPA whatsoever, and were compared with an equal-sized group consisting of the most active study participants (> or =25 min of LTPA per day). After adjusting for significant covariates, BMI (18.7%), %FM (19.4%), and C-reactive protein (143%) were all lower, and %FFM was higher (7.2%), in active participants (all p < or = 0.05). Ten percent of active participants vs. 33% of inactive participants were insulin resistant (p = 0.03). Waist circumference (17.6%) and systolic blood pressure (15.3%) were lower in active vs. inactive participants with paraplegia (both p < or = 0.05), but not tetraplegia. In conclusion, greater daily LTPA is associated with lower levels of selected CVD and type 2 diabetes risk factors in individuals living with SCI. Whether this relationship translates into a lower incidence of these chronic diseases has yet to be determined.


Two sources of mitochondrial NADPH in the yeast Saccharomyces cerevisiae.
Cells of the yeast Saccharomyces cerevisiae contain three NAD kinases; namely, cytosolic Utr1p, cytosolic Yef1p, and mitochondrial Pos5p. Previously, the NADH kinase reaction catalyzed by Pos5p, rather than the NAD kinase reaction followed by the NADP(+)-dependent dehydrogenase reaction, had been regarded as a critical source of mitochondrial NADPH, which plays vital roles in various mitochondrial functions. This study demonstrates that the mitochondrial NADH kinase reaction is dispensable as a source of mitochondrial NADPH and emphasizes the importance of the NAD kinase reaction, followed by the mitochondrial NADP(+)-dependent dehydrogenase reaction. Of the potential dehydrogenases (malic enzyme, Mae1p; isocitrate dehydrogenase, Idp1p; and acetaldehyde dehydrogenases, Ald4/5p), evidence is presented that acetaldehyde dehydrogenases, and in particular Ald4p, play a prominent role in generating mitochondrial NADPH in the absence of the NADH kinase reaction. The physiological significance of the mitochondrial NADH kinase reaction in the absence of Ald4p is also demonstrated. In addition, Pos5p is confirmed to have a considerably higher NADH kinase activity than NAD kinase activity. Taking these results together, it is proposed that there are two sources of mitochondrial NADPH in yeast: one is the mitochondrial Pos5p-NADH kinase reaction and the other is the mitochondrial Pos5p-NAD kinase reaction followed by the mitochondrial NADP(+)-dependent acetaldehyde dehydrogenase reaction.


Multi-isotope (carbon and chlorine) analysis for fingerprinting and site characterization at a fractured bedrock aquifer contaminated by chlorinated ethenes.
The use of compound specific multi-isotope approach (C and Cl) in the characterization of a chlorinated ethenes contaminated fractured aquifer allows the identification of several sources and contaminant plumes, as well as the occurrence of biodegradation and mixing processes. The study site is located in Spain with contamination resulting in groundwater concentrations of up to 50mg/L of trichloroethene (TCE), the most abundant chlorinated ethene, and 7 mg/L of tetrachloroethene (PCE). The potential sources of contamination including abandoned barrels, an underground tank, and a disposal lagoon, showed a wide range in δ(13)C values from -15.6 to -40.5‰ for TCE and from -18.5 to -32.4‰ for PCE, allowing the use of isotope fingerprinting for tracing of the origin and migration of these contaminants in the aquifer. In contrast, there is no difference between the δ(37)Cl values for TCE in the contaminant sources, ranging from +0.53 to +0.66‰. Variations of δ(37)Cl and δ(13)C in the different contaminant plumes were used to investigate the role of biodegradation in groundwater. Moreover, the isotopic data were incorporated into a reactive transport model for determination of whether the isotope pattern observed downstream from the tank's source could be explained by the simultaneous effect of mixing and biodegradation. The results demonstrate that a multi-isotope approach is a valuable tool for characterization of complex sites such as fractured bedrock aquifer contaminated by multiple sources, providing important information which can be used by consultants and site managers to prioritize and design more successful remediation strategies.


Clinical exome performance for reporting secondary genetic findings.
Reporting clinically actionable incidental genetic findings in the course of clinical exome testing is recommended by the American College of Medical Genetics and Genomics (ACMG). However, the performance of clinical exome methods for reporting small subsets of genes has not been previously reported. In this study, 57 exome data sets performed as clinical (n = 12) or research (n = 45) tests were retrospectively analyzed. Exome sequencing data was examined for adequacy in the detection of potentially pathogenic variant locations in the 56 genes described in the ACMG incidental findings recommendation. All exons of the 56 genes were examined for adequacy of sequencing coverage. In addition, nucleotide positions annotated in HGMD (Human Gene Mutation Database) were examined. The 56 ACMG genes have 18 336 nucleotide variants annotated in HGMD. None of the 57 exome data sets possessed a HGMD variant. The clinical exome test had inadequate coverage for >50% of HGMD variant locations in 7 genes. Six exons from 6 different genes had consistent failure across all 3 test methods; these exons had high GC content (76%-84%). The use of clinical exome sequencing for the interpretation and reporting of subsets of genes requires recognition of the substantial possibility of inadequate depth and breadth of sequencing coverage at clinically relevant locations. Inadequate depth of coverage may contribute to false-negative clinical exome results.


Cigarette smoking and pulmonary tuberculosis in northern California.
A positive association between smoking and increased risk of tuberculosis disease is well documented for populations outside the USA. However, it is unclear whether smoking increases risk of tuberculosis in the USA, where both smoking prevalence and disease rates are much lower than in the countries where previous studies have been conducted. To explore the tuberculosis-smoking association in a more generalisable US population-based sample, we conducted a nested case-control study among members of Kaiser Permanente Northern California (KPNC). We identified all newly diagnosed cases of active pulmonary tuberculosis (PTB) disease between 1996 and 2010. Each of the 2380 cases were individually matched to two controls by age, gender and race/ethnicity. ORs and 95% CIs for the association between smoking status and PTB were calculated using conditional logistic regression adjusted for all matching factors. Increased PTB risk was observed among ever-smokers (OR=1.35; 95% CI 1.19 to 1.53), as well as current (OR=1.26; 95% CI 1.08 to 1.48) and past (OR=1.43; 95% CI 1.23 to 1.67) smokers, compared with never-smokers. Increased intensity and duration of smoking were also positively associated with PTB risk. Our findings among a more generalisable US population support the hypothesis that smoking increases risk of PTB, underscoring the importance of tobacco cessation and prevention programmes in eliminating tuberculosis.


Relationships between Paraspinal Muscle Activity and Lumbar Inter-Vertebral Range of Motion.
Control of the lumbar spine requires contributions from both the active and passive sub-systems. Identifying interactions between these systems may provide insight into the mechanisms of low back pain. However, as a first step it is important to investigate what is normal. The purpose of this study was to explore the relationships between the lumbar inter-vertebral range of motion and paraspinal muscle activity during weight-bearing flexion in healthy controls using quantitative fluoroscopy (QF) and surface electromyography (sEMG). Contemporaneous lumbar sEMG and QF motion sequences were recorded during controlled active flexion of 60° using electrodes placed over Longissimus thoracis pars thoracis (TES), Longissimus thoracis pars lumborum (LES), and Multifidus (LMU). Normalised root mean square (RMS) sEMG amplitude data were averaged over five epochs, and the change in amplitude between epochs was calculated. The sEMG ratios of LMU/LES LMU/TES and LES/TES were also determined. QF was used to measure the maximum inter-vertebral range of motion from L2-S1, and correlation coefficients were calculated between sEMG amplitude variables and these measurements. Intra- and inter-session sEMG amplitude repeatability was also assessed for all three paraspinal muscles. The sEMG amplitude measurements were highly repeatable, and sEMG amplitude changes correlated significantly with L4-5 and L5-S1 IV-RoMmax (r = -0.47 to 0.59). The sEMG amplitude ratio of LES/TES also correlated with L4-L5 IV-RoMmax (r = -0.53). The relationships found may be important when considering rehabilitation for low back pain.


Crystal Structure of Aldehyde Dehydrogenase 16 Reveals Trans-Hierarchical Structural Similarity and a New Dimer.
The aldehyde dehydrogenase (ALDH) superfamily is a vast group of enzymes that catalyze the NAD+-dependent oxidation of aldehydes to carboxylic acids. ALDH16 is perhaps the most enigmatic member of the superfamily, owing to its extra C-terminal domain of unknown function and the absence of the essential catalytic cysteine residue in certain non-bacterial ALDH16 sequences. Herein we report the first production of recombinant ALDH16, the first biochemical characterization of ALDH16, and the first crystal structure of ALDH16. Recombinant expression systems were generated for the bacterial ALDH16 from Loktanella sp. and human ALDH16A1. Four high-resolution crystal structures of Loktanella ALDH16 were determined. Loktanella ALDH16 is found to be a bona fide enzyme, exhibiting NAD+-binding, ALDH activity, and esterase activity. In contrast, human ALDH16A1 apparently lacks measurable aldehyde oxidation activity, suggesting that it is a pseudoenzyme, consistent with the absence of the catalytic Cys in its sequence. The fold of ALDH16 comprises three domains: NAD+-binding, catalytic, and C-terminal. The latter is unique to ALDH16 and features a Rossmann fold connected to a protruding β-flap. The tertiary structural interactions of the C-terminal domain mimic the quaternary structural interactions of the classic ALDH superfamily dimer, a phenomenon we call "trans-hierarchical structural similarity." ALDH16 forms a unique dimer in solution, which mimics the classic ALDH superfamily dimer-of-dimer tetramer. Small-angle X-ray scattering shows that human ALDH16A1 has the same dimeric structure and fold as Loktanella ALDH16. We suggest that the Loktanella ALDH16 structure may be considered to be the archetype of the ALDH16 family.


Mesenchymal stem cell injection ameliorates chronic renal failure in a rat model.
CKD (chronic kidney disease) has become a public health problem. The therapeutic approaches have been able to reduce proteinuria, but have not been successful in limiting disease progression. In this setting, cell therapies associated with regenerative effects are attracting increasing interest. We evaluated the effect of MSC (mesenchymal stem cells) on the progression of CKD and the expression of molecular biomarkers associated with regenerative effects. Adult male Sprague-Dawley rats subjected to 5/6 NPX (nephrectomy) received a single intravenous infusion of 0.5×106 MSC or culture medium. A sham group subjected to the same injection was used as the control. Rats were killed 5 weeks after MSC infusion. Dye tracking of MSC was followed by immunofluorescence analysis. Kidney function was evaluated using plasma creatinine. Structural damage was evaluated by H&E (haematoxylin and eosin) staining, ED-1 abundance (macrophages) and interstitial α-SMA (α-smooth muscle actin). Repairing processes were evaluated by functional and structural analyses and angiogenic/epitheliogenic protein expression. MSC could be detected in kidney tissues from NPX animals treated with intravenous cell infusion. This group presented a marked reduction in plasma creatinine levels and damage markers ED-1 and α-SMA (P<0.05). In addition, treated rats exhibited a significant induction in epitheliogenic [Pax-2, bFGF (basic fibroblast growth factor) and BMP-7 (bone morphogenetic protein-7)] and angiogenic [VEGF (vascular endothelial growth factor) and Tie-2] proteins. The expression of these biomarkers of regeneration was significantly related to the increase in renal function. Many aspects of the cell therapy in CKD remain to be investigated in more detail: for example, its safety, low cost and the possible need for repeated cell injections over time. Beyond the undeniable importance of these issues, what still needs to be clarified is whether MSC administration has a real effect on the treatment of this pathology. It is precisely to this point that the present study aims to contribute.


Toward a Better Understanding of Adherence to Micronutrient Powders: Generating Theories to Guide Program Design and Evaluation Based on a Review of Published Results.
Background: The Global Alliance for Improved Nutrition is conducting theory-driven process evaluations of micronutrient powder (MNP) programs. Objective: The aim was to generate preliminary theories about factors affecting adherence to recommendations with regard to point-of-use fortification of foods with MNPs. Methods: A literature search was conducted to identify documents with content related to adherence to MNPs as an intervention provided at home to children 6-59 mo of age. Thirty-five studies and 6 program descriptions were identified. We used thematic analyses to generate a comprehensive list of factors that could influence adherence, followed by content analysis to quantify the results. We developed a Program Impact Pathway to concretize the points at which the factors identified affect the process of adherence. Results: In the set of documents reviewed (n = 41), the most influential factors, measured by number of documents reporting the factor having effect, were 1) caregivers' perception of positive changes as a result of MNP use (n = 14), 2) caregivers' perceived child acceptance of food with MNPs (n = 12), and 3) caregivers' forgetfulness (n = 11). Behavior change communication channels (n = 13) and messages (n = 12) were the most frequently reported program design features influencing caregiver knowledge and subsequent adherence. Administration regimen (n = 10), which may be related to caregivers' capacity to remember to give MNPs, was also a frequently cited program design feature affecting adherence. Conclusions: The preponderance of knowledge and perception factors may reflect an underlying theoretical bias among researchers as to what they measure. To achieve programs that support greater adherence, we need to adopt a cultural-ecological perspective to inform program design in order to address a broader set of determinants. Studies that assess progress across the impact pathway, particularly from adherence to biological outcomes, would also provide guidance for evaluation studies, particularly when time or other constraints limit the potential to measure biological outcomes.


Standard Peripheral Blood Mononuclear Cell Cryopreservation Selectively Decreases Detection of Nine Clinically Relevant T Cell Markers.
Biobanking is an operational component of various epidemiological studies and clinical trials. Although peripheral blood is routinely acquired and stored in biobanks, the effects of specimen processing on cell composition and clinically relevant functional markers of T cells still require a systematic evaluation. In this study, we assessed 25 relevant T cell markers in human PBMCs and showed that the detection of nine membrane markers (e.g., PD-1, CTLA4, KLRG1, CD25, CD122, CD127, CCR7, and others reflecting exhaustion, senescence, and other functions) was reduced among at least one T cell subset following standard processing, although the frequency of CD4, CD8, and regulatory T cells was unaffected. Nevertheless, a 6-mo-long cryopreservation did not impair the percentages of cells expressing many other membrane and all the eight tested intracellular lineage or functional T cell markers. Our findings uncover that several clinically relevant markers are particularly affected by processing and the interpretation of those results in clinical trials and translational research should be done with caution.


Real-time, high-resolution imaging of tumor cells in genetically engineered and orthotopic models of thyroid cancer.
Genetically engineered and orthotopic xenograft mouse models have been instrumental for increasing our understanding of thyroid cancer progression and for the development of novel therapeutic approaches in a setting that is more physiologically relevant than the classical subcutaneous flank implants. However, the anatomical location of the thyroid gland precludes a non-invasive analysis at the cellular level of the interactions between tumor cells and the surrounding microenvironment and does not allow a real-time evaluation of the response of tumor cells to drug treatments. As a consequence, such studies have generally only relied on endpoint approaches, limiting the amount and depth of the information that could be gathered. Here we describe the development of an innovative approach to imaging specific aspects of thyroid cancer biology, based on the implantation of a permanent, minimally invasive optical window that allows high-resolution, multi-day, intravital imaging of the behavior and cellular dynamics of thyroid tumors in the mouse. We show that this technology allows visualization of fluorescently tagged tumor cells both in immunocompetent, genetically engineered mouse models of anaplastic thyroid cancer (ATC) and in immunocompromised mice carrying orthotopic implanted human or mouse ATC cells. Furthermore, the use of recipient mice in which endothelial cells and macrophages are fluorescently labeled allows the detection of the spatial and functional relationship between tumor cells and their microenvironment. Finally, we show that ATC cells expressing a fluorescent biosensor for caspase 3 activity can be effectively utilized to evaluate, in real-time, the efficacy and kinetics of action of novel small molecule therapeutics. This novel approach to intravital imaging of thyroid cancer represents a platform that will allow, for the first time, the longitudinal, in situ analysis of tumor cell responses to therapy and of their interaction with the microenvironment.


Independent regulation of nifHDK operon transcription and DNA rearrangement during heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120.
The filamentous cyanobacterium Anabaena sp. strain PCC 7120 expresses the genes required for nitrogen fixation in terminally differentiated cells called heterocysts. The nifHDK operon encodes the nitrogenase polypeptides and is expressed at high levels in heterocysts. During heterocyst differentiation, an 11-kb DNA element is excised from the nifD gene by site-specific recombination. The xisA gene, located on the 11-kb element, is required for the excision of the element. Transcription and DNA rearrangement of the nifHDK operon both occur late during heterocyst differentiation, about 18 to 24 h after induction, suggesting that the regulation of these events might be coupled. We show that heterocyst-specific transcription and DNA rearrangement of the nifHDK operon are independent of one another. Northern (RNA) analysis of the xisA mutant strain DW12-2.2, which cannot excise the nifD 11-kb element or fix nitrogen, showed that the nifH and nifD genes are transcribed on unrearranged chromosomes. The nifK gene was not transcribed in DW12-2.2, indicating that its expression is dependent on the nifH promoter and excision of the 11-kb element from the operon. A 1.68-kb DNA fragment containing the nifH promoter was deleted from the chromosome to produce the mutant strain LW1. LW1 formed heterocysts but did not grow on nitrogen-free medium and showed no transcription through nifD. Southern analysis of LW1 showed normal excision of the 11-kb element from the nifHDK operon, indicating that transcription from the nifH promoter is not required for the developmentally regulated DNA rearrangement.


Contemporary cocaine use patterns and associated harms in Melbourne and Sydney, Australia.
The aim of this paper was to explore the nature of cocaine use and harms through a cross-sectional survey of cocaine users interviewed in the two largest Australian cities of Sydney (n = 88) and Melbourne (n = 77) between October 2004 and January 2005. The study supported previous findings that Australian cocaine users could be classified broadly into two types. The majority of cocaine users interviewed were classified as socially and economically integrated. They were young, employed, well-educated people who generally snorted cocaine on a recreational basis, typically in conjunction with other illicit and licit drugs. A second group of socially and economically marginalised users, residing mainly in Sydney, injected cocaine often in conjunction with heroin. This group reported significantly higher levels of cocaine use, cocaine dependence, criminal behaviour and human immunodeficiency virus (HIV) risk-taking behaviour. Heroin use was found to predict independently higher levels of cocaine use, criminal behaviour, needle sharing and physical problems in this sample, suggesting that increased resources and coverage for combined heroin/cocaine users may have scope for reducing cocaine-related problems in the Australian community.