Dissociable neuroanatomical correlates of subsecond and suprasecond time perception.
The ability to estimate durations varies across individuals. Although previous studies have reported that individual differences in perceptual skills and cognitive capacities are reflected in brain structures, it remains unknown whether timing abilities are also reflected in the brain anatomy. Here, we show that individual differences in the ability to estimate subsecond and suprasecond durations correlate with gray matter (GM) volume in different parts of cortical and subcortical areas. Better ability to discriminate subsecond durations was associated with a larger GM volume in the bilateral anterior cerebellum, whereas better performance in estimating the suprasecond range was associated with a smaller GM volume in the inferior parietal lobule. These results indicate that regional GM volume is predictive of an individual's timing abilities. These morphological results support the notion that subsecond durations are processed in the motor system, whereas suprasecond durations are processed in the parietal cortex by utilizing the capacity of attention and working memory to keep track of time.


Emerging adults' expectations for pornography use in the context of future committed romantic relationships: a qualitative study.
Using qualitative content analysis from the written comments of 404 primarily heterosexual college students, we examined (1) their expectations for pornography use while married or in a committed long-term relationship and (2) variations by gender. Four prominent groups emerged. A majority of men (70.8 %) and almost half of women (45.5 %) reported circumstances (alone or with their partners) wherein pornography use was acceptable in a relationship and several conditions for, and consequences associated with, such use also emerged. Another group (22.3 % men; 26.2 % women) viewed pornography use as unacceptable because of being in a committed relationship whereas a third group (5.4 % men; 12.9 % women) reported that pornography use was unacceptable in any context or circumstance. A final group emerged of a few women (10.4 %) who stated that a partner's use of pornography was acceptable, but they did not expect to use it personally. Implications for relationship education among emerging adults and future research on pornography use within the context of romantic relationships are discussed.


ProPhylo: partial phylogenetic profiling to guide protein family construction and assignment of biological process.
Phylogenetic profiling is a technique of scoring co-occurrence between a protein family and some other trait, usually another protein family, across a set of taxonomic groups. In spite of several refinements in recent years, the technique still invites significant improvement. To be its most effective, a phylogenetic profiling algorithm must be able to examine co-occurrences among protein families whose boundaries are uncertain within large homologous protein superfamilies. Partial Phylogenetic Profiling (PPP) is an iterative algorithm that scores a given taxonomic profile against the taxonomic distribution of families for all proteins in a genome. The method works through optimizing the boundary of each protein family, rather than by relying on prebuilt protein families or fixed sequence similarity thresholds. Double Partial Phylogenetic Profiling (DPPP) is a related procedure that begins with a single sequence and searches for optimal granularities for its surrounding protein family in order to generate the best query profiles for PPP. We present ProPhylo, a high-performance software package for phylogenetic profiling studies through creating individually optimized protein family boundaries. ProPhylo provides precomputed databases for immediate use and tools for manipulating the taxonomic profiles used as queries. ProPhylo results show universal markers of methanogenesis, a new DNA phosphorothioation-dependent restriction enzyme, and efficacy in guiding protein family construction. The software and the associated databases are freely available under the open source Perl Artistic License from ftp://ftp.jcvi.org/pub/data/ppp/.


An Introduction to Propensity Score Methods for Reducing the Effects of Confounding in Observational Studies.
The propensity score is the probability of treatment assignment conditional on observed baseline characteristics. The propensity score allows one to design and analyze an observational (nonrandomized) study so that it mimics some of the particular characteristics of a randomized controlled trial. In particular, the propensity score is a balancing score: conditional on the propensity score, the distribution of observed baseline covariates will be similar between treated and untreated subjects. I describe 4 different propensity score methods: matching on the propensity score, stratification on the propensity score, inverse probability of treatment weighting using the propensity score, and covariate adjustment using the propensity score. I describe balance diagnostics for examining whether the propensity score model has been adequately specified. Furthermore, I discuss differences between regression-based methods and propensity score-based methods for the analysis of observational data. I describe different causal average treatment effects and their relationship with propensity score analyses.


The evolutionary genetics of emerging plant RNA viruses.
Over the years, agriculture across the world has been compromised by a succession of devastating epidemics caused by new viruses that spilled over from reservoir species or by new variants of classic viruses that acquired new virulence factors or changed their epidemiological patterns. Viral emergence is usually associated with ecological change or with agronomical practices bringing together reservoirs and crop species. The complete picture is, however, much more complex, and results from an evolutionary process in which the main players are ecological factors, viruses' genetic plasticity, and host factors required for virus replication, all mixed with a good measure of stochasticity. The present review puts emergence of plant RNA viruses into the framework of evolutionary genetics, stressing that viral emergence begins with a stochastic process that involves the transmission of a preexisting viral strain into a new host species, followed by adaptation to the new host.


Neferine induces autophagy-dependent cell death in apoptosis-resistant cancers via ryanodine receptor and Ca2+-dependent mechanism.
Resistance of cancer cells to chemotherapy is a significant clinical concern and mechanisms regulating cell death in cancer therapy, including apoptosis, autophagy or necrosis, have been extensively investigated over the last decade. Accordingly, the identification of medicinal compounds against chemoresistant cancer cells via new mechanism of action is highly desired. Autophagy is important in inducing cell death or survival in cancer therapy. Recently, novel autophagy activators isolated from natural products were shown to induce autophagic cell death in apoptosis-resistant cancer cells in a calcium-dependent manner. Therefore, enhancement of autophagy may serve as additional therapeutic strategy against these resistant cancers. By computational docking analysis, biochemical assays, and advanced live-cell imaging, we identified that neferine, a natural alkaloid from Nelumbo nucifera, induces autophagy by activating the ryanodine receptor and calcium release. With well-known apoptotic agents, such as staurosporine, taxol, doxorubicin, cisplatin and etoposide, utilized as controls, neferine was shown to induce autophagic cell death in a panel of cancer cells, including apoptosis-defective and -resistant cancer cells or isogenic cancer cells, via calcium mobilization through the activation of ryanodine receptor and Ulk-1-PERK and AMPK-mTOR signaling cascades. Taken together, this study provides insights into the cytotoxic mechanism of neferine-induced autophagy through ryanodine receptor activation in resistant cancers.


Examining the "evolution of increased competitive ability" hypothesis in response to parasites and pathogens in the invasive paper wasp Polistes dominula.
Successful invaders often become established in new ranges by outcompeting native species. The "evolution of increased competitive ability" hypothesis predicts that invasive species are subjected to less predation and parasitization than sympatric native species, and thus can allocate resources from defence and immunity to growth and fecundity, thereby achieving higher fitness. In this study, we examined whether American invasive Polistes dominula paper wasps have reduced immunocompetence. To explore this scenario, we tested their susceptibility towards parasites and pathogens at both the individual (immune defence) and colony levels, i.e. hygienic behaviour (removal of diseased individuals by nestmates). First, we examined the response to the specific coevolved parasite Xenos vesparum (lost after invasion) in terms of individual host susceptibility and hygienic behaviour. Second, we explored the response against general pathogens by quantifying the bacterial clearance in individual wasps after a challenge with Escherichia coli and hygienic behaviour after a challenge with the fungus Beauveria bassiana. Our results show that American invasive P. dominula have a higher response against X. vesparum at the colony level, but at the individual level their susceptibility is not significantly different from conspecifics of the native range. On the other hand, invasive P. dominula display lower response after a challenge with general pathogens at both the individual and colony levels. While supporting the hypothesis of a reduction of immunocompetence towards general pathogens in invasive species, these findings also suggest that the response against coevolved parasites might follow different evolutionary pathways which are not always easily predictable.


Prevention of Alzheimer's disease pathology by cannabinoids: neuroprotection mediated by blockade of microglial activation.
Alzheimer's disease (AD) is characterized by enhanced beta-amyloid peptide (betaA) deposition along with glial activation in senile plaques, selective neuronal loss, and cognitive deficits. Cannabinoids are neuroprotective agents against excitotoxicity in vitro and acute brain damage in vivo. This background prompted us to study the localization, expression, and function of cannabinoid receptors in AD and the possible protective role of cannabinoids after betaA treatment, both in vivo and in vitro. Here, we show that senile plaques in AD patients express cannabinoid receptors CB1 and CB2, together with markers of microglial activation, and that CB1-positive neurons, present in high numbers in control cases, are greatly reduced in areas of microglial activation. In pharmacological experiments, we found that G-protein coupling and CB1 receptor protein expression are markedly decreased in AD brains. Additionally, in AD brains, protein nitration is increased, and, more specifically, CB1 and CB2 proteins show enhanced nitration. Intracerebroventricular administration of the synthetic cannabinoid WIN55,212-2 to rats prevent betaA-induced microglial activation, cognitive impairment, and loss of neuronal markers. Cannabinoids (HU-210, WIN55,212-2, and JWH-133) block betaA-induced activation of cultured microglial cells, as judged by mitochondrial activity, cell morphology, and tumor necrosis factor-alpha release; these effects are independent of the antioxidant action of cannabinoid compounds and are also exerted by a CB2-selective agonist. Moreover, cannabinoids abrogate microglia-mediated neurotoxicity after betaA addition to rat cortical cocultures. Our results indicate that cannabinoid receptors are important in the pathology of AD and that cannabinoids succeed in preventing the neurodegenerative process occurring in the disease.


Benign thyroid adenomas among persons X-irradiated in infancy for enlarged thymus glands.
Thyroid adenoma incidence in a cohort of 2657 infants given X-ray treatment for a supposedly enlarged thymus gland, along with 4833 unirradiated siblings, has been ascertained for an average of 37 years since irradiation. Estimated thyroid doses ranged from 0.03 to > 8 Gy, with 62% receiving < 0.5 Gy. After excluding 4 adenoma cases with concurrent or previous thyroid cancer, there were 86 cases with pathologically diagnosed thyroid adenomas in the irradiated group and 11 in the sibling controls. The estimated excess relative risk (ERR) was 6.3 per gray (90% CI = 3.7, 11.2). Once the dose group with > or = 6 Gy, which was producing downward curvature in the dose-response function, was removed, the curve was compatible with linearity and the ERR was 7.8 per gray. Thyroid adenoma rates were elevated even at low doses: the lowest dose group (< 0.25 Gy) showed a significant elevation in risk. The relative risk appeared to be constant over time and was comparable for both sexes. Excess adenoma risk was observed in the irradiated group to the maximum follow-up interval of about 50 years. A number of potential risk factors for thyroid adenoma were examined both as risk factors in their own right and as modifiers of the radiogenic risk. Parity and use of hormones in relation to menopause were significantly associated with thyroid adenoma risk in women, while education, Jewish origin, history of hyperthyroidism or hypothyroidism, and family history of cancer were also adenoma risk factors in both sexes. An examination of interactions between possible risk factors and radiation suggested that women with a history of oral contraceptive use or hysterectomy and persons with a family history of cancer may have greater risk (per unit dose) of radiogenic thyroid adenomas than their counterparts.


Adenovirus-mediated human tissue kallikrein gene delivery inhibits neointima formation induced by interruption of blood flow in mice.
Tissue kallikrein cleaves kininogen to produce vasoactive kinin peptides. Binding of kinins to bradykinin B(2) receptors on vascular endothelial cells stimulates the release of nitric oxide and prostacyclin, thus activating the cGMP and cAMP pathways. In this study, we evaluated the effects of adenovirus-mediated human tissue kallikrein gene (Ad.CMV-cHK) delivery in a mouse model of arterial remodeling induced by permanent alteration in shear stress conditions. Mice underwent ligature of the left common carotid artery and were injected intravenously with saline or 1.8 x 10(9) plaque-forming units of Ad.CMV-cHK or control virus (Ad.CMV-LacZ). Fourteen days after surgery, morphometric analysis revealed that Ad. CMV-cHK reduced neointima formation by 52% (P<0.05) compared with Ad. CMV-LacZ. Expression of human tissue kallikrein (HK) mRNA was detected in mouse carotid artery, aorta, kidney, heart, and liver, and recombinant HK was present in the urine and plasma of mice receiving HK gene. Kallikrein gene transfer resulted in increases in urinary kinin, cGMP, and cAMP levels. The protective action of Ad. CMV-cHK on neointima formation was significantly reduced (P<0.05) in mice with knockout of the kinin B(2) receptor gene compared with wild-type control mice (J129Sv mice). In contrast, the effect of Ad. CMV-cHK was amplified (P<0.05) in transgenic mice overexpressing human B(2) receptor compared with wild-type control mice (c57/Bl6 mice). Thus, the inhibitory effect of recombinant kallikrein on structural alterations caused by the interruption of blood flow appears to be mediated by the B(2) receptor. These results provide new insight into the role of the tissue kallikrein-kinin system in vascular remodeling and suggest the application of HK gene therapy to treat restenosis and atherosclerosis.


Cell cycle inhibitors for the treatment of NSCLC.
Lung cancer remains to be the leading cause of cancer-related death worldwide. Treatment of lung cancer still poses a significant challenge. Cell cycle is a tightly integrated process and is frequently aberrant in lung cancer. Cell cycle inhibitors have emerged as novel therapeutics, in anticipation of overcoming the unrestricted cell division and growth in lung cancer. In this article, we first address the potential roles of cell cycle proteins and cell cycle deregulation in the development of lung cancer. The review then provides an overview for several major categories of cell cycle inhibitors with particular attention to their tolerability and disease control in early phases of lung cancer trials. Targeted agents against different components of cell cycle regulation, such as cyclin-dependent kinase, polo-like kinase, checkpoint kinase and aurora kinase, are currently in clinical development for lung cancer management. Their clinical benefits remain to be defined. When evaluated as single agents in lung cancer, cell cycle inhibitors are often associated with limited clinical activity and tolerable toxicities. The key challenges in the drug development are to understand resistance mechanisms and to identify predictive biomarkers that can potentially guide patient selection and optimize the utility of these targeted inhibitors.


mTOR and S6K1 mediate assembly of the translation preinitiation complex through dynamic protein interchange and ordered phosphorylation events.
In response to nutrients, energy sufficiency, hormones, and mitogenic agents, S6K1 phosphorylates several targets linked to translation. However, the molecular mechanisms whereby S6K1 is activated, encounters substrate, and contributes to translation initiation are poorly understood. We show that mTOR and S6K1 maneuver on and off the eukaryotic initiation factor 3 (eIF3) translation initiation complex in a signal-dependent, choreographed fashion. When inactive, S6K1 associates with the eIF3 complex, while the S6K1 activator mTOR/raptor does not. Cell stimulation promotes mTOR/raptor binding to the eIF3 complex and phosphorylation of S6K1 at its hydrophobic motif. Phosphorylation results in S6K1 dissociation, activation, and subsequent phosphorylation of its translational targets, including eIF4B, which is then recruited into the complex in a phosphorylation-dependent manner. Thus, the eIF3 preinitiation complex acts as a scaffold to coordinate a dynamic sequence of events in response to stimuli that promote efficient protein synthesis.


Characterization and Preoperative Risk Analysis of Leiomyosarcomas at a High-Volume Tertiary Care Center.
Uterine morcellation in minimally invasive surgery has recently come under scrutiny because of inadvertent dissemination of malignant tissue, including leiomyosarcomas commonly mistaken for fibroids. Identification of preoperative risk factors is crucial to ensure that oncologic care is delivered when suspicion for malignancy is high, while offering minimally invasive hysterectomies to the remaining patients. The aim of this study was to characterize risk factors for uterine leiomyosarcomas by reviewing preoperative, intraoperative, and postoperative data with an emphasis on the presence of concurrent fibroids. A retrospective case-control study of women undergoing hysterectomy with pathologic diagnosis of uterine leiomyosarcoma at a tertiary care center between January 2005 and April 2014. Thirty-one women were identified with leiomyosarcoma and matched to 124 controls. Cases with leiomyosarcoma were more likely to have undergone menopause and to present with larger uteri (19- vs 9-week sized), with the most common presenting complaint being a pelvic mass (35.5% vs 8.9%). Controls were ten times more likely to have undergone a tubal ligation (30.6% vs 3.2%). Endometrial sampling detected malignancy preoperatively in only 50% of cases. Leiomyosarcomas were more commonly present when pelvic masses were identified in addition to fibroids on preoperative imaging. Most leiomyosarcoma cases (77.4%) were performed by oncologists via an abdominal approach (83.9%), with only 2 of 31 leiomyosarcomas being morcellated. Comparative analysis of preoperative imaging and postoperative pathology showed that in patients with leiomyosarcoma, fibroids were misdiagnosed 58.1% of the time, and leiomyosarcomas arose directly from fibroids in only 6.5% of cases. Leiomyosarcoma risk factors include older age/postmenopausal status, enlarged uteri of greater than 10 weeks, and lack of previous tubal ligation. Preoperative testing failed to definitively identify leiomyosarcomas, although the presence of synchronous pelvic masses in fibroid uteri should raise clinical suspicion. Given the difficulty of preoperative identification, future efforts should focus on the development of safer minimally invasive techniques for uterine morcellation.


Vitamin K and the prevention of fractures: systematic review and meta-analysis of randomized controlled trials.
Observational and some experimental data suggest that low intake of vitamin K may be associated with an increased risk of fracture. To assess whether oral vitamin K (phytonadione and menaquinone) supplementation can reduce bone loss and prevent fractures. The search included the following electronic databases: MEDLINE (1966 to June 2005), EMBASE (1980 to June 2005), the Cochrane Library (issue 2, 2005), the ISI Web of Science (1945 to June 2005), the National Research Register (inception to the present), Current Controlled Trials, and the Medical Research Council Research Register. Randomized controlled trials that gave adult participants oral phytonadione and menaquinone supplements for longer than 6 months were included in this review. Four authors extracted data on changes in bone density and type of fracture. All articles were double screened and double data extracted. Thirteen trials were identified with data on bone loss, and 7 reported fracture data. All studies but 1 showed an advantage of phytonadione and menaquinone in reducing bone loss. All 7 trials that reported fracture effects were Japanese and used menaquinone. Pooling the 7 trials with fracture data in a meta-analysis, we found an odds ratio (OR) favoring menaquinone of 0.40 (95% confidence interval [CI], 0.25-0.65) for vertebral fractures, an OR of 0.23 (95% CI, 0.12-0.47) for hip fractures, and an OR of 0.19 (95% CI, 0.11-0.35) for all nonvertebral fractures. This systematic review suggests that supplementation with phytonadione and menaquinone-4 reduces bone loss. In the case of the latter, there is a strong effect on incident fractures among Japanese patients.


Gender Differences in Posttraumatic Stress Disorder and Help Seeking in the U.S. Army.
Inconsistent findings between studies of gender differences in mental health outcomes in military samples have left open questions of differential prevalence in posttraumatic stress disorder (PTSD) among all United States Army soldiers and in differential psychosocial and comorbid risk and protective factor profiles and their association with receipt of treatment. This study assesses the prevalence and risk factors of screening positive for PTSD for men and women based on two large, population-based Army samples obtained as part of the 2005 and 2008 U.S. Department of Defense Surveys of Health Related Behaviors among Active Duty Military Personnel. The study showed that overall rates of PTSD, as measured by several cutoffs of the PTSD Checklist, are similar between active duty men and women, with rates increasing in both men and women between the two study time points. Depression and problem alcohol use were strongly associated with a positive PTSD screen in both genders, and combat exposure was significantly associated with a positive PTSD screen in men. Overall, active duty men and women who met criteria for PTSD were equally likely to receive mental health counseling or treatment, though gender differences in treatment receipt varied by age, race, social support (presence of spouse at duty station), history of sexual abuse, illness, depression, alcohol use, and combat exposure. The study demonstrates that the prevalence of PTSD as well as the overall utilization of mental health services is similar for active duty men compared with women. However, there are significant gender differences in predictors of positive PTSD screens and receipt of PTSD treatment.


Early and Long-term Outcomes of Older Adults after Acute Care Encounters for Chronic Obstructive Pulmonary Disease Exacerbation.
Older patients are at high risk of death and rehospitalization after hospitalizations for acute exacerbations of chronic obstructive pulmonary disease (AE-COPD). Emergency department visits comprise a substantial portion of acute care encounters in this patient population. The risks of mortality and repeat acute care encounters, including both hospital readmission and repeat emergency department visits, after AE-COPD among older adults are not well understood. To examine early and long-term rates of death and repeat acute care encounters after hospitalization or emergency department visit for AE-COPD in Medicare fee-for-service beneficiaries and to identify patient characteristics, including medical comorbid conditions, associated with these outcomes. A retrospective analysis was conducted using a nationally representative 5% sample of Medicare fee-for-service claims data from the U.S. Centers for Medicare and Medicaid Services to identify Medicare beneficiaries 65 years or older who had an acute care episode for an AE-COPD between January 1, 2006, and December 31, 2010 (n = 52,741). Outcomes of interest were all-cause mortality, repeat acute care encounters for any cause, and repeat acute care encounters for AE-COPD at 30 days, 1 year, and 3 years. Acute care encounters, including hospitalizations and emergency department visits for AE-COPD, were associated with substantial subsequent mortality risk, with 4.6, 24.4, and 48.2% dying by 30 days, 1 year, and 3 years, respectively. The risk of repeat hospitalization or emergency department visit was similarly high, with 1 in 4 patients having a repeat acute care encounter within 30 days of discharge, increasing to 9 in 10 in the next 3 years. Several comorbid conditions and other patient factors, including heart failure, malnutrition, dual eligibility for Medicare and Medicaid, and prior supplemental oxygen use, were independently associated with increased risk of repeat acute care encounter. Repeat hospitalizations and emergency department visits and death are common in older fee-for-service Medicare beneficiaries seen in acute care for AE-COPD. Our results suggest that addressing important comorbid conditions, such as heart failure or malnutrition, and targeting resources to oxygen-dependent or dual Medicare- and Medicaid-eligible patients may help modify these outcomes.


The amylase inhibitor montbretin A reveals a new glycosidase inhibition motif.
The complex plant flavonol glycoside montbretin A is a potent (Ki = 8 nM) and specific inhibitor of human pancreatic α-amylase with potential as a therapeutic for diabetes and obesity. Controlled degradation studies on montbretin A, coupled with inhibition analyses, identified an essential high-affinity core structure comprising the myricetin and caffeic acid moieties linked via a disaccharide. X-ray structural analyses of the montbretin A-human α-amylase complex confirmed the importance of this core structure and revealed a novel mode of glycosidase inhibition wherein internal π-stacking interactions between the myricetin and caffeic acid organize their ring hydroxyls for optimal hydrogen bonding to the α-amylase catalytic residues D197 and E233. This novel inhibitory motif can be reproduced in a greatly simplified analog, offering potential for new strategies for glycosidase inhibition and therapeutic development.


State variation in primary care physician supply: implications for health reform Medicaid expansions.
Under the Patient Protection and Affordable Care Act (PPACA), Medicaid enrollment is expected to grow by 16 million people by 2019, an increase of more than 25 percent. Given the unwillingness of many primary care physicians (PCPs) to treat new Medicaid patients, policy makers and others are concerned about adequate primary care capacity to meet the increased demand. States with the smallest number of PCPs per capita overall--generally in the South and Mountain West--potentially will see the largest percentage increases in Medicaid enrollment, according to a new national study by the Center for Studying Health System Change (HSC). In contrast, states with the largest number of PCPs per capita--primarily in the Northeast--will see more modest increases in Medicaid enrollment. Moreover, geographic differences in PCP acceptance of new Medicaid patients reflect differences in overall PCP supply, not geographic differences in PCPs' willingness to treat Medicaid patients. The law also increases Medicaid reimbursement rates for certain services provided by primary care physicians to 100 percent of Medicare rates in 2013 and 2014. However, the reimbursement increases are likely to have the greatest impact in states that already have a large number of PCPs accepting Medicaid patients. In fact, the percent increase of PCPs accepting Medicaid patients in these states is likely to exceed the percent increase of new Medicaid enrollees. The reimbursement increases will have much less impact in states with a relatively small number of PCPs accepting Medicaid patients now because many of these states already reimburse primary care at rates close to or exceeding 100 percent of Medicare. As a result, growth in Medicaid enrollment in these states will greatly outpace growth in the number of primary care physicians willing to treat new Medicaid patients.


Experimental quantification of long distance dispersal potential of aquatic snails in the gut of migratory birds.
Many plant seeds and invertebrates can survive passage through the digestive system of birds, which may lead to long distance dispersal (endozoochory) in case of prolonged retention by moving vectors. Endozoochorous dispersal by waterbirds has nowadays been documented for many aquatic plant seeds, algae and dormant life stages of aquatic invertebrates. Anecdotal information indicates that endozoochory is also possible for fully functional, active aquatic organisms, a phenomenon that we here address experimentally using aquatic snails. We fed four species of aquatic snails to mallards (Anas platyrhynchos), and monitored snail retrieval and survival over time. One of the snail species tested was found to survive passage through the digestive tract of mallards as fully functional adults. Hydrobia (Peringia) ulvae survived up to five hours in the digestive tract. This suggests a maximum potential transport distance of up to 300 km may be possible if these snails are taken by flying birds, although the actual dispersal distance greatly depends on additional factors such as the behavior of the vectors. We put forward that more organisms that acquired traits for survival in stochastic environments such as wetlands, but not specifically adapted for endozoochory, may be sufficiently equipped to successfully pass a bird's digestive system. This may be explained by a digestive trade-off in birds, which maximize their net energy intake rate rather than digestive efficiency, since higher efficiency comes with the cost of prolonged retention times and hence reduces food intake. The resulting lower digestive efficiency allows species like aquatic snails, and potentially other fully functional organisms without obvious dispersal adaptations, to be transported internally. Adopting this view, endozoochorous dispersal may be more common than up to now thought.


An efficient gene disruption system for the nematophagous fungus Purpureocillium lavendulum.
The fungus Purpureocillium lavendulum (formally Paecilomyces lilacinus) is a natural enemy of insects and plant-parasitic nematodes, and has been used as an important bio-control agent against agricultural pests all over the world. In order to understand the genetic mechanisms governing its biocontrol efficiency and other biological processes, an effective gene disruption system is needed. Here we report the development of an efficient system which integrates selective markers that differ from Purpureocillium lilacinum, a one-step construction method for gene knockout plasmids, and a ku80 knockout strain for efficient homologous recombination. With this system, we effectively disrupted the transcription factors in the central regulation pathway of sporulation and a serine protease which were contributed to nematode infection, demonstrating this system as an efficient gene disrupting system for further characterization of genes involved in the development and pathogenesis of this fungus.


Ever-increasing Caesarean section and its economic burden in Bangladesh.
Cesarean Section (CS) delivery has been increasing rapidly worldwide and Bangladesh is no exception. In Bangladesh, the CS rate has increased from about 3% in 2000 to about 24% in 2014. This study examines trend in CS in Bangladesh over the last fifteen years and implications of this increasing CS rates on health care expenditures. Birth data from Bangladesh Demographic and Health Survey (BDHS) for the years 2000-2014 have been used for the trend analysis and 2010 Bangladesh Maternal Mortality Survey (BMMS) data were used for estimating health care expenditure associated with CS. Although the share of institutional deliveries increased four times over the years 2000 to 2014, the CS deliveries increased eightfold. In 2000, only 33% of institutional deliveries were conducted through CS and the rate increased to 63% in 2014. Average medical care expenditure for a CS delivery in Bangladesh was about BDT 22,085 (USD 276) in 2010 while the cost of a normal delivery was BDT 3,565 (USD 45). Health care expenditure due to CS deliveries accounted for about 66.5% of total expenditure on all deliveries in Bangladesh in 2010. About 10.3% of Total Health Expenditure (THE) in 2010 was due to delivery costs, while CS costs contribute to 6.9% of THE and rapid increase in CS deliveries will mean that delivering babies will represent even a higher proportion of THE in the future despite declining crude birth rate. High CS delivery rate and the negative health outcomes associated with the procedure on mothers and child births incur huge economic burden on the families. This is creating inappropriate allocation of scarce resources in the poor economy like Bangladesh. Therefore it is important to control this unnecessary CS practices by the health providers by introducing litigation and special guidelines in the health policy.


The Role of Transient Receptor Potential Melastatin 7 (TRPM7) in Cell Viability: A Potential Target to Suppress Breast Cancer Cell Cycle.
The divalent cation-selective channel transient receptor potential melastatin 7 (TRPM7) channel was shown to affect the proliferation of some types of cancer cell. However, the function of TRPM7 in the viability of breast cancer cells remains unclear. Here we show that TRPM inhibitors suppressed the viability of TRPM7-expressing breast cancer cells. We first demonstrated that the TRPM7 inhibitors 2-aminoethyl diphenylborinate (2-APB), ginsenoside Rd (Gin Rd), and waixenicin A preferentially suppressed the viability of human embryonic kidney HEK293 overexpressing TRPM7 (HEK-M7) cells over wildtype HEK293 (WT-HEK). Next, we confirmed the effects of 2-APB on the TRPM7 channel functions by whole-cell currents and divalent cation influx. The inhibition of the viability of HEK-M7 cells by 2-APB was not mediated by the increase in cell death but by the interruption of the cell cycle. Similar to HEK-M7 cells, the viability of TRPM7-expressing human breast cancer MDA-MB-231, AU565, and T47D cells were also suppressed by 2-APB by arresting the cell cycle in the S phase. Furthermore, in a novel TRPM7 knock-out MDA-MB-231 (KO-231) cell line, decreased divalent influx and reduced proliferation were observed compared to the wildtype MDA-MB-231 cells. 2-APB and Gin Rd preferentially suppressed the viability of wildtype MDA-MB-231 cells over KO-231 by affecting the cell cycle in wildtype but not KO-231 cells. Our results suggest that TRPM7 regulates the cell cycle of breast cancers and is a potential therapeutic target.


Improved tissue culture conditions for engineered skeletal muscle sheets.
The potential clinical utility of engineered muscle is currently restricted by limited in vitro capacity of expanded muscle precursor cells to fuse and form mature myofibers. The purpose of this study was to use isotropic skeletal muscle sheets to explore the impact of (1) fibroblast coculture and (2) fibroblast-conditioned media (fCM) on in vitro myogenesis. Muscle sheets were prepared by seeding varying ratios of skeletal myoblasts and fibroblasts on a biomimetic substrate and culturing the resulting tissue in either control media or fCM. Muscle sheets were prepared from two cell subpopulations, (1) C2C12 and NOR-10 and (2) primary neonatal rat skeletal muscle cells (nSKM). In C2C12/Nor-10 muscle sheets fCM conferred a myogenic advantage early in culture; at D1 a statistically significant 3.12 ± 0.8-fold increase in myofiber density was observed with fCM. A high purity satellite cell population was collected from an initially mixed population of nSKMs via cell sorting for positive α 7-integrin expression. On D6, tissue sheets with low fibroblast concentrations (0 & 10%) cultured in fCM had increased average myofiber density (4.8 ± 0.2 myofibers/field) compared to tissue sheets with high fibroblast concentrations (50%) cultured in control media (1.0 ± 0.1 myofibers/field). Additionally, fCM promoted longer, thicker myofibers with a mature phenotype.


Local field potential recordings in a non-human primate model of Parkinsons disease using the Activa PC + S neurostimulator.
Using the Medtronic Activa® PC + S system, this study investigated how passive joint manipulation, reaching behavior, and deep brain stimulation (DBS) modulate local field potential (LFP) activity in the subthalamic nucleus (STN) and globus pallidus (GP). Five non-human primates were implanted unilaterally with one or more DBS leads. LFPs were collected in montage recordings during resting state conditions and during motor tasks that facilitate the expression of parkinsonian motor signs. These recordings were made in the naïve state in one subject, in the parkinsonian state in two subjects, and in both naïve and parkinsonian states in two subjects. LFPs measured at rest were consistent over time for a given recording location and parkinsonian state in a given subject; however, LFPs were highly variable between subjects, between and within recording locations, and across parkinsonian states. LFPs in both naïve and parkinsonian states across all recorded nuclei contained a spectral peak in the beta band (10-30 Hz). Moreover, the spectral content of recorded LFPs was modulated by passive and active movement of the subjects' limbs. LFPs recorded during a cued-reaching task displayed task-related beta desynchronization in STN and GP. The bidirectional capabilities of the Activa® PC + S also allowed for recording LFPs while delivering DBS. The therapeutic effect of STN DBS on parkinsonian rigidity outlasted stimulation for 30-60 s, but there was no correlation with beta band power. This study emphasizes (1) the variability in spontaneous LFPs amongst subjects and (2) the value of using the Activa® PC + S system to record neural data in the context of behavioral tasks that allow one to evaluate a subject's symptomatology.


First record of atypical pigmentation pattern in fin whale Balaenoptera physalus in the Atlantic Ocean.
Atypical pigmentation, which is rarely observed in the wild, may influence social interactions between animals and can be detrimental for survival. Hypopigmentation, which is the lack of pigment in a part or on the entire body, is a type of atypical pigmentation pattern that can be either acquired (e.g. vitiligo) or congenital resulting from the inheritance of mutations in pigment-related genes (e.g. albinism, leucism and piebaldism). This study documents atypical pigmentation in a fin whale Balaenoptera physalus off the northwestern coast of the Iberian Peninsula (Atlantic Ocean). Photographic and video data collected between 2016 and 2017 on 30 individual fin whales were examined. One fully-grown fin whale exhibited hypopigmentation. Several white patches of different shapes and sizes were present across the body of the fin whale including on the head, body, dorsal fin, flippers, and flukes. The position, shape, and lack of inflammation of the white patches on the whale observed, along with its body length and condition, might indicate that the depigmentation pattern is due to vitiligo. To our knowledge, this is the first case of atypical pigmentation pattern in fin whales described with photographs and video records. As these observations are rare, especially in highly migratory, long-lived, marine mammal species, this study provides valuable information to better understand the occurrence of this phenomenon. Further studies are needed to determine the ecological and physiological implications of atypical colourations, which might have a significant influence on the animal's survival.


Slum Health: Arresting COVID-19 and Improving Well-Being in Urban Informal Settlements.
The informal settlements of the Global South are the least prepared for the pandemic of COVID-19 since basic needs such as water, toilets, sewers, drainage, waste collection, and secure and adequate housing are already in short supply or non-existent. Further, space constraints, violence, and overcrowding in slums make physical distancing and self-quarantine impractical, and the rapid spread of an infection highly likely. Residents of informal settlements are also economically vulnerable during any COVID-19 responses. Any responses to COVID-19 that do not recognize these realities will further jeopardize the survival of large segments of the urban population globally. Most top-down strategies to arrest an infectious disease will likely ignore the often-robust social groups and knowledge that already exist in many slums. Here, we offer a set of practice and policy suggestions that aim to (1) dampen the spread of COVID-19 based on the latest available science, (2) improve the likelihood of medical care for the urban poor whether or not they get infected, and (3) provide economic, social, and physical improvements and protections to the urban poor, including migrants, slum communities, and their residents, that can improve their long-term well-being. Immediate measures to protect residents of urban informal settlements, the homeless, those living in precarious settlements, and the entire population from COVID-19 include the following: (1) institute informal settlements/slum emergency planning committees in every urban informal settlement; (2) apply an immediate moratorium on evictions; (3) provide an immediate guarantee of payments to the poor; (4) immediately train and deploy community health workers; (5) immediately meet Sphere Humanitarian standards for water, sanitation, and hygiene; (6) provide immediate food assistance; (7) develop and implement a solid waste collection strategy; and (8) implement immediately a plan for mobility and health care. Lessons have been learned from earlier pandemics such as HIV and epidemics such as Ebola. They can be applied here. At the same time, the opportunity exists for public health, public administration, international aid, NGOs, and community groups to innovate beyond disaster response and move toward long-term plans.


UNC-4/UNC-37-dependent repression of motor neuron-specific genes controls synaptic choice in Caenorhabditis elegans.
The UNC-4 homeoprotein and the Groucho-like corepressor UNC-37 specify synaptic choice in the Caenorhabditis elegans motor neuron circuit. In unc-4 mutants, VA motor neurons are miswired with inputs from interneurons normally reserved for their lineal sisters, the VB motor neurons. Here we show that UNC-4 and UNC-37 function together in VA motor neurons to repress VB-specific genes and that this activity depends on physical contact between UNC-37 and a conserved Engrailed-like repressor domain (eh1) in UNC-4. Missense mutations in the UNC-4 eh1 domain disrupt interactions between UNC-4 and UNC-37 and result in the loss of UNC-4-dependent repressor activity in vivo. A compensatory amino acid substitution in UNC-37 suppresses specific unc-4 alleles by restoring physical interactions with UNC-4 as well as UNC-4-dependent repression of VB-specific genes. We propose that repression of VB-specific genes by UNC-4 and UNC-37 is necessary for the creation of wild-type inputs to VA motor neurons. The existence of mammalian homologs of UNC-4 and UNC-37 indicates that a similar mechanism could regulate synaptic choice in the vertebrate spinal cord.


An extremely robust strong-field control of atomic coherence.
We propose and analyze a scheme for adiabatic creation of maximum coherence of 0.5 with a controllable phase between a ground state and the excited state in a model Λ-structured atom using two short laser pulses. One of the pulses has constant carrier frequency quasi-resonant with transition between an initially empty ground and the excite states. The frequency of the second pulse is swept through the resonance with the adjacent transition between the initially populated ground state and the common excited state of the atom. We demonstrate high degree of robustness of the scheme against variation of parameters of the laser radiation in relatively broad region of values. The proposed scheme may find practical applications in the field of multi-photon ionization, high-order harmonics and Raman sideband generation, as well as in nonlinear wave mixing in coherently prepared media.


HIV infection in patients attending clinics for sexually transmitted diseases in England and Wales 1988. A Collaborative Study by Consultants in Genitourinary Medicine and the Public Health Laboratory Service.
A national study of the prevalence of HIV antibody in homosexual and heterosexual patients attending sexually transmitted disease clinics in the years 1985-7 was continued in 1988. Among homosexual men in two clinics in south east England HIV antibody was less prevalent in 1988 (30 (7.1%) of 420) than in 1987 (81 (14.6%) of 556) but was more prevalent in 1988 in other regions where the corresponding findings were 48 (4.9%) of 975 and 36 (2.8%) of 1291. HIV antibody prevalence continued to be higher in homosexual men aged 25 or more years or with one or more specified minor complaints. The trend was similar in heterosexual patients: among men in the south east in 1988 HIV antibody was found in 1 (0.1%) of 948 but in 10 (1.0%) of 962 in 1987 and among women in 1 (0.1%) of 1043 in 1988 but in 7 (0.7%) of 949 in 1987. In other regions the corresponding findings for men were 2 (0.1%) of 5620 in 1988 and 3 (0.1%) of 5312 in 1987 and for women were 7 (0.2%) of 4483 in 1988 and 1 (0.1%) of 4778 in 1987. The two most notable differences between findings in 1987 and 1988 were that in the south east the number of heterosexual intravenous drug abusers with HIV antibody decreased from five to one among men and from five to none among women and for the first time in the study HIV antibody positive women without known risk factors were identified--1 (0.1%) of 1043 in the south east and 2 (0.04%) of 4483 in other regions. Refusals to participate continued to increase but, as before, comparison of patients who agreed and refused in terms of age, the presence of symptoms suggesting HIV infection, travel abroad, and number of sexual partners in the past twelve months showed little evidence of selective bias that might have artificially reduced the prevalence.


Early marginal ulcer following Roux-en-Y gastric bypass under proton pump inhibitor treatment: prospective multicentric study.
Causal factors of gastrojejunal ulcers after Roux-en-Y gastric bypass include peptic acid secretion from the gastric pouch. Esomeprazole is a potent inhibitor of acid secretion. To assess the occurrence of dyspepsia and gastrojejunal ulcers within the first 2 months after Roux-en-Y gastric bypass during the use of esomeprazole. One hundred eighteen morbid obese subjects were submitted to Roux-en-Y gastric bypass. Preoperative upper gastrointestinal tract endoscopy was negative for H. pylori. All subjects received esomeprazole for 60 days after surgery. Two weeks after surgery only 13 mild symptoms were reported. After 2 months, 17 also moderate complaints were registered. Endoscopy around the 60th day showed esophagitis in 10 (8.5%), hiatal hernia in 2 (1.7%), foreign body in the anastomotic line in 12 (10.2%) and gastrojejunal ulcers was observed in 9 (7.6%) subjects, 2 of which had a suture material or metallic staple granuloma in the gastrojejunostomy. Ten subjects took nonsteroidal anti-inflammatory drugs at least once during study, but none of them developed ulcer. None of the subjects with ulcer had dyspeptic symptoms. The incidence of ulcer in the gastrojejunal anastomosis within the first 2 months following Rouxen-Y gastric bypass under proton pump inhibitors is considerable. It was not related to the use of non-steroidal anti-inflammatory drugs, highlighting the possibility of ischemia and foreign body as causal factors. The ulcers were asymptomatic, and all post-surgical dyspeptic symptoms were moderate in severity.


Pupillary response to representations of light in paintings.
It is known that, although the level of light is the primary determinant of pupil size, cognitive factors can also affect pupil diameter. It has been demonstrated that photographs of the sun produce pupil constriction independently of their luminance and other low-level features, suggesting that high-level visual processing may also modulate pupil response. Here, we measure pupil response to artistic paintings of the sun, moon, or containing a uniform lighting, that, being mediated by the artist's interpretation of reality and his technical rendering, require an even higher level of interpretation compared with photographs. We also study how chromatic content and spatial layout affect the results by presenting grey-scale and inverted versions of each painting. Finally, we assess directly with a categorization test how subjective image interpretation affects pupil response. We find that paintings with the sun elicit a smaller pupil size than paintings with the moon, or paintings containing no visible light source. The effect produced by sun paintings is reduced by disrupting contextual information, such as by removing color or manipulating the relations between paintings features that make more difficult to identify the source of light. Finally, and more importantly, pupil diameter changes according to observers' interpretation of the scene represented in the same stimulus. In conclusion, results show that the subcortical pupillary response to light is modulated by subjective interpretation of luminous objects, suggesting the involvement of cortical systems in charge of cognitive processes, such as attention, object recognition, familiarity, memory, and imagination.


A slow cooling rate of indomethacin melt spatially confined in microcontainers increases the physical stability of the amorphous drug without influencing its biorelevant dissolution behaviour.
Amorphous indomethacin was prepared by melting the γ-form of indomethacin, spatially confined within microcontainers (inner diameter of 223 μm), followed by cooling of the melt at a rate of 14, 23 or 36 K/min. The physical stability of the amorphous indomethacin within microcontainers was investigated using Raman microscopy. Furthermore, the dissolution behaviour of confined amorphous indomethacin was evaluated in biorelevant intestinal media at pH 6.5. After 30 days of storage, 10.3 ± 1.2 % of the amorphous indomethacin cooled at 14 K/min and confined within microcontainers was found to be crystalline. When the melt of indomethacin was cooled at 23 or 36 K/min, 20.7 ± 1.5 and 31.0 ± 2.6 % of the indomethacin were found to be crystalline after storage for 30 days. Scanning electron microscopy showed a smooth surface of amorphous indomethacin within the microcontainers when cooling the melt at 14 K/min, whereas cracks and an uneven surface were observed when cooling at rates of 23 and 36 K/min. The uneven surface is hypothesised to be the main reason for the lower physical stability, as the cracks could act as nucleation sites for crystal growth. The rate of cooling was not seen to have any effect on the dissolution of amorphous indomethacin from the microcontainers.


Can altered muscle synergies control unimpaired gait?
Recent studies have postulated that the human motor control system recruits groups of muscles through low-dimensional motor commands, or muscle synergies. This scheme simplifies the neural control problem associated with the high-dimensional structure of the neuromuscular system. Several lines of evidence have suggested that neurological injuries, such as stroke or cerebral palsy, may reduce the dimensions that are available to the motor control system, and these altered dimensions or synergies are thought to contribute to impaired walking patterns. However, no study has investigated whether impaired low-dimensional control spaces necessarily lead to impaired walking patterns. In this study, using a two-dimensional model of walking, we developed a synergy-based control framework that can simulate the dynamics of walking. The simulation analysis showed that a synergy-based control scheme can produce well-coordinated movements of walking matching unimpaired gait. However, when the dimensions available to the controller were reduced, the simplified emergent pattern deviated from unimpaired gait. A system with two synergies, similar to those seen after neurological injury, could not produce an unimpaired walking pattern. These findings provide further evidence that altered muscle synergies can contribute to impaired gait patterns and may need to be directly addressed to improve gait after neurological injury.


Injection seeding for the enhancement of high-order anti-Stokes stimulated Raman scattering.
We have investigated the effects of Stokes and anti-Stokes pulse injection on high-order anti-Stokes Raman scattering for efficient generation of vacuum-ultraviolet radiation. The output energies as well as the spatial profiles of anti-Stokes pulses were much improved by injection. Enhancement factors increased for higher orders of anti-Stokes pulses. The shift of optimum pressure together with the change of profile from a ring to a Gaussian-like shape induced by injection indicates that the enhancement is due to the modification of phase-matching geometry in four-wave Raman mixing for anti-Stokes generation.


Constructing hierarchical task analysis in surgery.
Generic technical skills are required by a surgeon to perform a complete operation or procedure. They alone do not form a task or subtask but allow the surgeon to perform so. Specific technical skills are required to complete a task or subtask, which can be depicted by hierarchical task analysis (HTA). In this study we aim to demonstrate a reliable and valid method to construct a surgical HTA. One hundred thirty video recordings of operations and procedures (30 laparoscopic cholecystectomies, 20 open inguinal hernia repairs, 20 saphenofemoral junction ligations, 20 upper GI and 40 lower GI endoscopies) from 37 different expert surgeons were assessed in view of constructing a HTA. Three research surgeons with more than eight years of postgraduate surgical experience assessed each operation or procedure blindly and independently and constructed a HTA. Each consultant surgeon assessed the HTA constructed by the researchers and modified it according to his/her own technical style. For tasks there was a 100% correlation between the researchers and individual expert surgeons. Mean interrater reliability for subtasks was k = 0.89 (range = 0.81-0.95), p < 0.05. Content and face validities of the HTA were confirmed by the expert surgeons. This study outlines a valid and reliable method of constructing a surgical task analysis and HTA for any operation or procedure, which could be used to assess and evaluate trainees' and expert surgeons' specific technical skills.


Development of a Job-Exposure Matrix (AsbJEM) to Estimate Occupational Exposure to Asbestos in Australia.
Occupational exposure data on asbestos are limited and poorly integrated in Australia so that estimates of disease risk and attribution of disease causation are usually calculated from data that are not specific for local conditions. To develop a job-exposure matrix (AsbJEM) to estimate occupational asbestos exposure levels in Australia, making optimal use of the available exposure data. A dossier of all available exposure data in Australia and information on industry practices and controls was provided to an expert panel consisting of three local industrial hygienists with thorough knowledge of local and international work practices. The expert panel estimated asbestos exposures for combinations of occupation, industry, and time period. Intensity and frequency grades were estimated to enable the calculation of annual exposure levels for each occupation-industry combination for each time period. Two indicators of asbestos exposure intensity (mode and peak) were used to account for different patterns of exposure between occupations. Additionally, the probable type of asbestos fibre was determined for each situation. Asbestos exposures were estimated for 537 combinations of 224 occupations and 60 industries for four time periods (1943-1966; 1967-1986; 1987-2003; ≥2004). Workers in the asbestos manufacturing, shipyard, and insulation industries were estimated to have had the highest average exposures. Up until 1986, 46 occupation-industry combinations were estimated to have had exposures exceeding the current Australian exposure standard of 0.1 f ml(-1). Over 90% of exposed occupations were considered to have had exposure to a mixture of asbestos varieties including crocidolite. The AsbJEM provides empirically based quantified estimates of asbestos exposure levels for Australian jobs since 1943. This exposure assessment application will contribute to improved understanding and prediction of asbestos-related diseases and attribution of disease causation.


Managers' practices related to work-family balance predict employee cardiovascular risk and sleep duration in extended care settings.
An increasing proportion of U.S. workers have family caregiving responsibilities. The purpose of this study was to determine whether employees in extended care settings whose managers are supportive, open, and creative about work-family needs, such as flexibility with work schedules, have lower cardiovascular disease (CVD) risk and longer sleep than their less supported counterparts. From semistructured interviews with managers, we constructed a work-family balance score of manager openness and creativity in dealing with employee work-family needs. Trained interviewers collected survey and physiologic outcome data from 393 employees whose managers had a work-family score. Employee outcomes are sleep duration (actigraphy) and CVD risk assessed by blood cholesterol, high glycosylated hemoglobin/diabetes, blood pressure/hypertension, body-mass index, and tobacco consumption. Employees whose managers were less supportive slept less (29 min/day) and were over twice as likely to have 2 or more CVD risk factors (ORs = 2.1 and 2.03 for low and middle manager work-family scores, respectively) than employees whose managers were most open and creative. Employees who provide direct patient care exhibited particularly elevated CVD risk associated with low manager work-family score. Managers' attitudes and practices may affect employee health, including sleep duration and CVD risk.


Iron and zinc content of selected foods in the diet of schoolchildren in Kumi district, east of Uganda: a cross-sectional study.
Iron and zinc are essential micronutrients for humans and deficiency of the two elements is widespread in the world with the highest prevalence in less developed countries. There are few data on dietary intake of iron and zinc in Uganda, and no food composition table is available. There is hardly any widely published literature that clearly documents the quality of Ugandan children's diet. Thus information of both food intake and the concentration of these trace elements in local food ingredients are needed in order to assess daily intake. The present study focused on the iron and zinc content in selected foods and intake of the micronutrients iron and zinc among schoolchildren in Kumi District, Uganda. Over a period of 4 weeks single 24-hour dietary recall interviews were carried out on a convenience sample of 178 schoolchildren (9-15 years old). Data from the dietary recalls was used when selecting foods for chemical analysis. Results from this study showed that the iron concentrations varied, and were high in some cereals and vegetables. The zinc concentrations in foods generally corresponded with results from other African countries (Mali and Kenya). Data from the 24-hour dietary recall showed that the daily Recommended Nutrient Intake (RNI) was met for iron but not for zinc. The schoolchildren of Kumi district had a predominantly vegetable based diet. Foods of animal origin were consumed occasionally. The iron content in the selected foods was high and variable, and higher than in similar ingredients from Kenya and Mali, while the zinc concentrations were generally in accordance with reported values. The total daily zinc (mg) intake does not meet the daily RNI. The iron intake is adequate according to RNI, but due to iron contamination and reduced bioavailability, RNI may not be met in a vegetable based diet. More studies are needed to investigate possible sources of contamination.


Cell-Type-Specific Sensorimotor Processing in Striatal Projection Neurons during Goal-Directed Behavior.
Goal-directed sensorimotor transformation drives important aspects of mammalian behavior. The striatum is thought to play a key role in reward-based learning and action selection, receiving glutamatergic sensorimotor signals and dopaminergic reward signals. Here, we obtain whole-cell membrane potential recordings from the dorsolateral striatum of mice trained to lick a reward spout after a whisker deflection. Striatal projection neurons showed strong task-related modulation, with more depolarization and action potential firing on hit trials compared to misses. Direct pathway striatonigral neurons, but not indirect pathway striatopallidal neurons, exhibited a prominent early sensory response. Optogenetic stimulation of direct pathway striatonigral neurons, but not indirect pathway striatopallidal neurons, readily substituted for whisker stimulation evoking a licking response. Our data are consistent with direct pathway striatonigral neurons contributing a "go" signal for goal-directed sensorimotor transformation leading to action initiation. VIDEO ABSTRACT.


Production and characterization of the slime polysaccharide of Pseudomonas aeruginosa.
The slime polysaccharides produced by Pseudomonas aeruginosa isolated from a variety of human infections were investigated. Slime production in culture seemed optimal when adequate amounts of carbohydrate were present and under conditions of either high osmotic pressure or inadequate protein supply. The polysaccharides produced by the organisms were similar to each other, to the slime of Azotobacter vinelandii, and to seaweed alginic acids. They were composed of beta-1,4-linked d-mannuronic acid residues and variable amounts of its 5-epimer l-guluronic acid. All bacterial polymers contained o-acetyl groups which are absent in the alginates. The polysaccharides differed considerably in the ratio of mannuronic to guluronic acid content and in the number of o-acetyl groups. The particular composition of the slime was not found to be characteristic for the disease process from which the mucoid variants of P. aeruginosa were obtained.


Decreases in colonic and systemic inflammation in chronic HIV infection after IL-7 administration.
Despite antiretroviral therapy (ART), some HIV-infected persons maintain lower than normal CD4(+) T-cell counts in peripheral blood and in the gut mucosa. This incomplete immune restoration is associated with higher levels of immune activation manifested by high systemic levels of biomarkers, including sCD14 and D-dimer, that are independent predictors of morbidity and mortality in HIV infection. In this 12-week, single-arm, open-label study, we tested the efficacy of IL-7 adjunctive therapy on T-cell reconstitution in peripheral blood and gut mucosa in 23 ART suppressed HIV-infected patients with incomplete CD4(+) T-cell recovery, using one cycle (consisting of three subcutaneous injections) of recombinant human IL-7 (r-hIL-7) at 20 µg/kg. IL-7 administration led to increases of both CD4(+) and CD8(+) T-cells in peripheral blood, and importantly an expansion of T-cells expressing the gut homing integrin α4β7. Participants who underwent rectosigmoid biopsies at study baseline and after treatment had T-cell increases in the gut mucosa measured by both flow cytometry and immunohistochemistry. IL-7 therapy also resulted in apparent improvement in gut barrier integrity as measured by decreased neutrophil infiltration in the rectosigmoid lamina propria 12 weeks after IL-7 administration. This was also accompanied by decreased TNF and increased FOXP3 expression in the lamina propria. Plasma levels of sCD14 and D-dimer, indicative of systemic inflammation, decreased after r-hIL-7. Increases of colonic mucosal T-cells correlated strongly with the decreased systemic levels of sCD14, the LPS coreceptor - a marker of monocyte activation. Furthermore, the proportion of inflammatory monocytes expressing CCR2 was decreased, as was the basal IL-1β production of peripheral blood monocytes. These data suggest that administration of r-hIL-7 improves the gut mucosal abnormalities of chronic HIV infection and attenuates the systemic inflammatory and coagulation abnormalities that have been linked to it.


Improving protein secondary structure prediction using a simple k-mer model.
Some first order methods for protein sequence analysis inherently treat each position as independent. We develop a general framework for introducing longer range interactions. We then demonstrate the power of our approach by applying it to secondary structure prediction; under the independence assumption, sequences produced by existing methods can produce features that are not protein like, an extreme example being a helix of length 1. Our goal was to make the predictions from state of the art methods more realistic, without loss of performance by other measures. Our framework for longer range interactions is described as a k-mer order model. We succeeded in applying our model to the specific problem of secondary structure prediction, to be used as an additional layer on top of existing methods. We achieved our goal of making the predictions more realistic and protein like, and remarkably this also improved the overall performance. We improve the Segment OVerlap (SOV) score by 1.8%, but more importantly we radically improve the probability of the real sequence given a prediction from an average of 0.271 per residue to 0.385. Crucially, this improvement is obtained using no additional information. http://supfam.cs.bris.ac.uk/kmer


Computer-Aided Discovery and Characterization of Novel Ebola Virus Inhibitors.
The Ebola virus (EBOV) causes severe human infection that lacks effective treatment. A recent screen identified a series of compounds that block EBOV-like particle entry into human cells. Using data from this screen, quantitative structure-activity relationship models were built and employed for virtual screening of a ∼17 million compound library. Experimental testing of 102 hits yielded 14 compounds with IC50 values under 10 μM, including several sub-micromolar inhibitors, and more than 10-fold selectivity against host cytotoxicity. These confirmed hits include FDA-approved drugs and clinical candidates with non-antiviral indications, as well as compounds with novel scaffolds and no previously known bioactivity. Five selected hits inhibited BSL-4 live-EBOV infection in a dose-dependent manner, including vindesine (0.34 μM). Additional studies of these novel anti-EBOV compounds revealed their mechanisms of action, including the inhibition of NPC1 protein, cathepsin B/L, and lysosomal function. Compounds identified in this study are among the most potent and well-characterized anti-EBOV inhibitors reported to date.


Accelerated aortic flow assessment with compressed sensing with and without use of the sparsity of the complex difference image.
Phase contrast (PC) cardiac MR is widely used for the clinical assessment of blood flow in cardiovascular disease. One of the challenges of PC cardiac MR is the long scan time which limits both spatial and temporal resolution. Compressed sensing reconstruction with accelerated PC acquisitions is a promising technique to increase the scan efficiency. In this study, we sought to use the sparsity of the complex difference of the two flow-encoded images as an additional constraint term to improve the compressed sensing reconstruction of the corresponding accelerated PC data acquisition. Using retrospectively under-sampled data, the proposed reconstruction technique was optimized and validated in vivo on 15 healthy subjects. Then, prospectively under-sampled data was acquired on 11 healthy subjects and reconstructed with the proposed technique. The results show that there is good agreement between the cardiac output measurements from the fully sampled data and the proposed compressed sensing reconstruction method using complex difference sparsity up to acceleration rate 5. In conclusion, we have developed and evaluated an improved reconstruction technique for accelerated PC cardiac MR that uses the sparsity of the complex difference of the two flow-encoded images.


Neuromodulation of pigment movement in the RPE of normal and 6-OHDA-lesioned goldfish retinas.
The role of dopamine as the endogenous signal-initiating light-dependent changes in the distribution of pigment granules in goldfish retinal pigment epithelium was investigated. In normal retinas, light adaptation resulted in the dispersion of pigment granules. This effect of light was mimicked by the intraocular injection of dopamine or serotonin, which is thought to increase endogenous dopamine release, into dark-adapted eyes. The effect of light, dopamine, or serotonin on dark-adapted retinas was blocked by the dopamine receptor antagonists haloperidol and sulpiride. However, lesioning the endogenous source of retinal dopamine, by prior intraocular injection of 6-hydroxydopamine (6-OHDA), did not block the dispersion of pigment granules in light-adapted retinas. No significant differences in pigment dispersion were noted between unlesioned and lesioned light- or dark-adapted retinas. However, the effect of light on pigment dispersion was no longer blocked by haloperidol or sulpiride in 6-OHDA lesioned animals. Dopamine and serotonin mimicked the effect of light when injected into lesioned dark-adapted eyes, but their effects were also not blocked by haloperidol or sulpiride. These results suggest that dopamine, acting on D2 receptors, is sufficient to induce pigment migration in unlesioned animals. In 6-OHDA-lesioned animals, however, pigment migration is mediated by a receptor mechanism other than D2.


A critical role for IL-17RB signaling in HTLV-1 tax-induced NF-κB activation and T-cell transformation.
Human T-cell leukemia virus type 1 (HTLV-1) infection is linked to the development of adult T-cell leukemia (ATL) and the neuroinflammatory disease HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP). The HTLV-1 Tax protein functions as a potent viral oncogene that constitutively activates the NF-κB transcription factor to transform T cells; however, the underlying mechanisms remain obscure. Here, using next-generation RNA sequencing we identified the IL-25 receptor subunit IL-17RB as an aberrantly overexpressed gene in HTLV-1 immortalized T cells. Tax induced the expression of IL-17RB in an IκB kinase (IKK) and NF-κB-dependent manner. Remarkably, Tax activation of the canonical NF-κB pathway in T cells was critically dependent on IL-17RB expression. IL-17RB and IL-25 were required for HTLV-1-induced immortalization of primary T cells, and the constitutive NF-κB activation and survival of HTLV-1 transformed T cells. IL-9 was identified as an important downstream target gene of the IL-17RB pathway that drives the proliferation of HTLV-1 transformed cells. Furthermore, IL-17RB was overexpressed in leukemic cells from a subset of ATL patients and also regulated NF-κB activation in some, but not all, Tax-negative ATL cell lines. Together, our results support a model whereby Tax instigates an IL-17RB-NF-κB feed-forward autocrine loop that is obligatory for HTLV-1 leukemogenesis.


Clinical subgroup clustering analysis in a systemic lupus erythematosus cohort from Western Pennsylvania.
Systemic lupus erythematosus (SLE) is a complex heterogenous autoimmune disease that can affect multiple organs. We performed clinical clustering analysis to describe a lupus cohort from the University of Pittsburgh Medical Center. A total of 724 patients who met the American College of Rheumatology (ACR) classification criteria for SLE were included in this study. Clustering was performed using the ACR classification criteria and the partitioning around medoid method. Correlation analysis was performed using the Spearman's Rho test. Patients with SLE in our cohort identify three district clinical disease subsets. Patients in cluster 1 were significantly more likely to develop renal and hematologic involvement, and had overrepresentation in African-American and male lupus patients. Clusters 2 and 3 identified a milder disease, with a significantly less likelihood of organ complications. Patients in cluster 2 are characterized by malar rash and photosensitivity, while patients in cluster 3 are characterized by oral ulcers, which is present in ~90% of patients within this cluster. The presence of photosensitivity or oral ulcers appears to be protective against the development of lupus nephritis in our cohort. We describe a large cohort of SLE from Western Pennsylvania and identify three distinct clinical disease subgroups. Clustering analysis might help to better manage and predict disease complications in heterogenous diseases like lupus.


Extraordinary low transmission effects for ultra-thin patterned metal films.
Thin metal films show a residual transmission for light in the visible and UV spectral range. This transmission can be strongly reduced by an appropriate sub-wavelength patterning of the metal film. Our investigation is focused on metal films with a thickness much below 100nm, where the transmission response is dominated by the individual posts acting like antennas and cannot be attributed to the excitation of surface plasmons. The almost complete suppression of transmission for ultra-thin metal films depends mainly on the absorber width, but not on the pitch of the pattern. The effect is robust with respect to imperfections of the geometry or larger features imprinted into the sub-wavelength pattern.


Inference of neuronal network spike dynamics and topology from calcium imaging data.
Two-photon calcium imaging enables functional analysis of neuronal circuits by inferring action potential (AP) occurrence ("spike trains") from cellular fluorescence signals. It remains unclear how experimental parameters such as signal-to-noise ratio (SNR) and acquisition rate affect spike inference and whether additional information about network structure can be extracted. Here we present a simulation framework for quantitatively assessing how well spike dynamics and network topology can be inferred from noisy calcium imaging data. For simulated AP-evoked calcium transients in neocortical pyramidal cells, we analyzed the quality of spike inference as a function of SNR and data acquisition rate using a recently introduced peeling algorithm. Given experimentally attainable values of SNR and acquisition rate, neural spike trains could be reconstructed accurately and with up to millisecond precision. We then applied statistical neuronal network models to explore how remaining uncertainties in spike inference affect estimates of network connectivity and topological features of network organization. We define the experimental conditions suitable for inferring whether the network has a scale-free structure and determine how well hub neurons can be identified. Our findings provide a benchmark for future calcium imaging studies that aim to reliably infer neuronal network properties.


Knowledge-fused differential dependency network models for detecting significant rewiring in biological networks.
Modeling biological networks serves as both a major goal and an effective tool of systems biology in studying mechanisms that orchestrate the activities of gene products in cells. Biological networks are context-specific and dynamic in nature. To systematically characterize the selectively activated regulatory components and mechanisms, modeling tools must be able to effectively distinguish significant rewiring from random background fluctuations. While differential networks cannot be constructed by existing knowledge alone, novel incorporation of prior knowledge into data-driven approaches can improve the robustness and biological relevance of network inference. However, the major unresolved roadblocks include: big solution space but a small sample size; highly complex networks; imperfect prior knowledge; missing significance assessment; and heuristic structural parameter learning. To address these challenges, we formulated the inference of differential dependency networks that incorporate both conditional data and prior knowledge as a convex optimization problem, and developed an efficient learning algorithm to jointly infer the conserved biological network and the significant rewiring across different conditions. We used a novel sampling scheme to estimate the expected error rate due to "random" knowledge. Based on that scheme, we developed a strategy that fully exploits the benefit of this data-knowledge integrated approach. We demonstrated and validated the principle and performance of our method using synthetic datasets. We then applied our method to yeast cell line and breast cancer microarray data and obtained biologically plausible results. The open-source R software package and the experimental data are freely available at http://www.cbil.ece.vt.edu/software.htm. Experiments on both synthetic and real data demonstrate the effectiveness of the knowledge-fused differential dependency network in revealing the statistically significant rewiring in biological networks. The method efficiently leverages data-driven evidence and existing biological knowledge while remaining robust to the false positive edges in the prior knowledge. The identified network rewiring events are supported by previous studies in the literature and also provide new mechanistic insight into the biological systems. We expect the knowledge-fused differential dependency network analysis, together with the open-source R package, to be an important and useful bioinformatics tool in biological network analyses.


Detection and correction for EPID and gantry sag during arc delivery using cine EPID imaging.
Electronic portal imaging devices (EPIDs) have been studied and used for pretreatment and in-vivo dosimetry applications for many years. The application of EPIDs for dosimetry in arc treatments requires accurate characterization of the mechanical sag of the EPID and gantry during rotation. Several studies have investigated the effects of gravity on the sag of these systems but each have limitations. In this study, an easy experiment setup and accurate algorithm have been introduced to characterize and correct for the effect of EPID and gantry sag during arc delivery. Three metallic ball bearings were used as markers in the beam: two of them fixed to the gantry head and the third positioned at the isocenter. EPID images were acquired during a 360° gantry rotation in cine imaging mode. The markers were tracked in EPID images and a robust in-house developed MATLAB code was used to analyse the images and find the EPID sag in three directions as well as the EPID + gantry sag by comparison to the reference gantry zero image. The algorithm results were then tested against independent methods. The method was applied to compare the effect in clockwise and counter clockwise gantry rotations and different source-to-detector distances (SDDs). The results were monitored for one linear accelerator over a course of 15 months and six other linear-accelerators from two treatment centers were also investigated using this method. The generalized shift patterns were derived from the data and used in an image registration algorithm to correct for the effect of the mechanical sag in the system. The Gamma evaluation (3%, 3 mm) technique was used to investigate the improvement in alignment of cine EPID images of a fixed field, by comparing both individual images and the sum of images in a series with the reference gantry zero image. The mechanical sag during gantry rotation was dependent on the gantry angle and was larger in the in-plane direction, although the patterns were not identical for various linear-accelerators. The reproducibility of measurements was within 0.2 mm over a period of 15 months. The direction of gantry rotation and SDD did not affect the results by more than 0.3 mm. Results of independent tests agreed with the algorithm within the accuracy of the measurement tools. When comparing summed images, the percentage of points with Gamma index <1 increased from 85.4% to 94.1% after correcting for the EPID sag, and to 99.3% after correction for gantry + EPID sag. The measurement method and algorithms introduced in this study use cine-images, are highly accurate, simple, fast, and reproducible. It tests all gantry angles and provides a suitable automatic analysis and correction tool to improve EPID dosimetry and perform comprehensive linac QA for arc treatments.


Adaptive capability of recurrent neural networks with fixed weights for series-parallel system identification.
By a fundamental neural filtering theorem, a recurrent neural network with fixed weights is known to be capable of adapting to an uncertain environment. This letter reports some mathematical results on the performance of such adaptation for series-parallel identification of a dynamical system as compared with the performance of the best series-parallel identifier possible under the assumption that the precise value of the uncertain environmental process is given. In short, if an uncertain environmental process is observable (not necessarily constant) from the output of a dynamical system or constant (not necessarily observable), then a recurrent neural network exists as a series-parallel identifier of the dynamical system whose output approaches the output of an optimal series-parallel identifier using the environmental process as an additional input.


Tests of gene order from three-locus linkage data.
Exact tests for gene order are derived and compared for three loci using linkage data from phase-known, completely informative marker loci (i.e. parents are heterozygotes with at most one allele identical at each locus), or from triple back-cross matings. A simulation method, based on resampling genotypes of children, is introduced to obtain approximations to the distribution of the test statistics for general mating types in families consisting of children and parents, with or without grandparents, as are used in many studies in human gene mapping. The method is illustrated by an application to linkage data on chromosome 13.


Contribution of polymorphisms in IKZF1 gene to childhood acute leukemia: a meta-analysis of 33 case-control studies.
Two common polymorphisms in the IKZF1 gene (rs4132601 and rs11978267 variants) have been reported to be associated with childhood acute leukemia (AL) risk, however the results were inconsistent. Here, we conducted a meta-analysis to generate large-scale evidence on whether IKZF1 variants are risk factors for childhood AL. The PubMed, Embase, EBSCO, and Web of Science were searched up to June 2, 2014 for studies on the association of IKZF1 polymorphisms with childhood AL risk. Data were extracted and the odd ratios (ORs) and 95% confidence intervals (95% CIs) were calculated by a fixed-effects or random-effects model. Subgroup analysis by ethnicity and leukemia subtype, sensitivity and cumulative meta-analyses were performed. Moreover, publication bias was assessed by Begg's and Egger's tests. In total, 33 case control studies were finally included in this meta-analysis. For rs4132601 polymorphism, significantly increased AL risk was observed in all genetic models (the association was still significant when the p value was Bonferroni adjusted to 0.025). In the subgroup analysis by tumor type, statistical association was observed in B-cell precursor ALL (BCP-ALL). Additionally, when stratified by ethnicity, significantly increased AL risk was only observed in European subgroup, but not among African or mixed population subgroups. Finally, similar results were found for rs11978267 polymorphism. In summary, this meta-analysis provides evidence that rs4132601 and rs11978267 polymorphisms in the IKZF1 gene might contribute to the occurrence of BCP-ALL, especially in European populations. Moreover, further studies with large sample size are required to clarify possible roles of IKZF1 variants in other ethnic groups (e.g., Asians and Africans).


Tobacco Smoking, Nicotine Dependence, and Patterns of Prescription Opioid Misuse: Results From a Nationally Representative Sample.
The misuse of prescription opioid medications is a growing public health crisis. Given evidence of complex nicotine-opioid interactions, and initial support for the role of smoking status as a risk factor for prescription opioid misuse, a more detailed analysis of how current and historical patterns of smoking may influence misuse of prescription opioids is warranted. The current study is the first to test whether varying levels of current/historical smoking (current daily, current intermittent, former daily, never) and indices of smoking heaviness/nicotine dependence may be associated with greater likelihood of past-year prescription opioid misuse in the general population. Data were derived from the National Survey on Drug Use and Health (N = 24,348). Consistent with hypotheses, after accounting for sociodemographic factors and major depressive/alcohol use disorders, both daily and intermittent smokers were greater than 3 times more likely to report past-year nonmedical prescription opioid use than were never smokers. In addition, daily smokers were observed to be nearly 5 times more likely, and intermittent smokers were nearly 3 times more likely, to have met past-year abuse/dependence criteria, relative to never smokers. Results further revealed positive associations between various indices of smoking heaviness/nicotine dependence and opioid medication misuse, and these findings remained largely consistent when analyses were stratified by gender. These findings indicate that smokers are not a homogeneous group with regard to risk for opioid misuse, and support the utility of comprehensive smoking assessment in the context of opioid-based treatment/tapering.


Genetic susceptibility to beryllium: a case-referent study of men and women of working age with sarcoidosis or other chronic lung disease.
The study was designed to investigate whether beryllium exposure was related to illness diagnosed as sarcoidosis. Chronic beryllium disease (CBD) and sarcoidosis are clinically and pathologically indistinguishable, with only the presence of beryllium-specific T-lymphocytes identifying CBD. Testing for such cells is not feasible in community studies of sarcoidosis but a second characteristic of CBD, its much greater incidence in those with a glutamic acid residue at position 69 of the HLA-DPB1 gene (Glu69), provides an alternative approach to answering this question. Cases of sarcoidosis aged 18-60 years diagnosed in Alberta, Canada, from 1999 to 2005 were approached through their specialist physician, together with age-matched and sex-matched referents with other chronic lung disease. Referents were grouped into chronic obstructive pulmonary disease (COPD), asthma and other lung disease. Participants completed a telephone questionnaire, including industry-specific questionnaires. DNA was extracted from mailed-in mouthwash samples and genotyped for Glu69. Duration of employment in types of work with independently documented beryllium exposure was calculated. DNA was extracted for 655 cases (270 Glu69 positive) and 1382 referents (561 positive). No increase in sarcoidosis was seen with either Glu69 or beryllium exposure (none, <10, ≥10 years) as main effects: longer duration in possible beryllium jobs was related to COPD. In Glu69 positive men with exposure ≥10 years, the trend towards increasing rate of COPD was reversed, and a significant interaction of duration of exposure and Glu69 was detected (OR=4.51 95% CI 1.17 to 17.48). The gene-environment interaction supports the hypothesis that some cases diagnosed as sarcoidosis result from occupational beryllium exposure.


Identification of three molecular and functional subtypes in canine hemangiosarcoma through gene expression profiling and progenitor cell characterization.
Canine hemangiosarcomas have been ascribed to an endothelial origin based on histologic appearance; however, recent findings suggest that these tumors may arise instead from hematopoietic progenitor cells. To clarify this ontogenetic dilemma, we used genome-wide expression profiling of primary hemangiosarcomas and identified three distinct tumor subtypes associated with angiogenesis (group 1), inflammation (group 2), and adipogenesis (group 3). Based on these findings, we hypothesized that a common progenitor may differentiate into the three tumor subtypes observed in our gene profiling experiment. To investigate this possibility, we cultured hemangiosarcoma cell lines under normal and sphere-forming culture conditions to enrich for tumor cell progenitors. Cells from sphere-forming cultures displayed a robust self-renewal capacity and exhibited genotypic, phenotypic, and functional properties consistent with each of the three molecular subtypes seen in primary tumors, including expression of endothelial progenitor cell (CD133 and CD34) and endothelial cell (CD105, CD146, and αvβ3 integrin) markers, expression of early hematopoietic (CD133, CD117, and CD34) and myeloid (CD115 and CD14) differentiation markers in parallel with increased phagocytic capacity, and acquisition of adipogenic potential. Collectively, these results suggest that canine hemangiosarcomas arise from multipotent progenitors that differentiate into distinct subtypes. Improved understanding of the mechanisms that determine the molecular and phenotypic differentiation of tumor cells in vivo could change paradigms regarding the origin and progression of endothelial sarcomas.


Semiparametric Regression Analysis of Panel Count Data: A Practical Review.
Panel count data arise in many applications when the event history of a recurrent event process is only examined at a sequence of discrete time points. In spite of the recent methodological developments, the availability of their software implementations has been rather limited. Focusing on a practical setting where the effects of some time-independent covariates on the recurrent events are of primary interest, we review semiparametric regression modelling approaches for panel count data that have been implemented in R package spef. The methods are grouped into two categories depending on whether the examination times are associated with the recurrent event process after conditioning on covariates. The reviewed methods are illustrated with a subset of the data from a skin cancer clinical trial.


Predicting objectively assessed physical activity from the content and regulation of exercise goals: evidence for a mediational model.
Grounded in self-determination theory (Deci & Ryan, 2000), the purpose of this work was to examine effects of the content and motivation of adults' exercise goals on objectively assessed moderate-to-vigorous physical activity (MVPA). After reporting the content and motivation of their exercise goals, 101 adult participants (Mage = 38.79 years; SD = 11.5) wore an ActiGraph (GT1M) accelerometer for seven days. Accelerometer data were analyzed to provide estimates of engagement in MVPA and bouts of physical activity. Goal content did not directly predict behavioral engagement; however, mediation analysis revealed that goal content predicted behavior via autonomous exercise motivation. Specifically, intrinsic versus extrinsic goals for exercise had a positive indirect effect on average daily MVPA, average daily MVPA accumulated in 10-min bouts and the number of days on which participants performed 30 or more minutes of MVPA through autonomous motivation. These results support a motivational sequence in which intrinsic versus extrinsic exercise goals influence physical activity behavior because such goals are associated with more autonomous forms of exercise motivation.


Supported Lipid Bilayers for the Generation of Dynamic Cell-Material Interfaces.
Supported lipid bilayers (SLB) offer unique possibilities for studying cellular membranes and have been used as a synthetic architecture to interact with cells. Here, the state-of-the-art in SLB-based technology is presented. The fabrication, analysis, characteristics and modification of SLBs are described in great detail. Numerous strategies to form SLBs on different substrates, and the means to patteren them, are described. The use of SLBs as model membranes for the study of membrane organization and membrane processes in vitro is highlighted. In addition, the use of SLBs as a substratum for cell analysis is presented, with discrimination between cell-cell and cell-extracellular matrix (ECM) mimicry. The study is concluded with a discussion of the potential for in vivo applications of SLBs.


p-i-n heterojunctions with BiFeO3 perovskite nanoparticles and p- and n-type oxides: photovoltaic properties.
We formed p-i-n heterojunctions based on a thin film of BiFeO3 nanoparticles. The perovskite acting as an intrinsic semiconductor was sandwiched between a p-type and an n-type oxide semiconductor as hole- and electron-collecting layer, respectively, making the heterojunction act as an all-inorganic oxide p-i-n device. We have characterized the perovskite and carrier collecting materials, such as NiO and MoO3 nanoparticles as p-type materials and ZnO nanoparticles as the n-type material, with scanning tunneling spectroscopy; from the spectrum of the density of states, we could locate the band edges to infer the nature of the active semiconductor materials. The energy level diagram of p-i-n heterojunctions showed that type-II band alignment formed at the p-i and i-n interfaces, favoring carrier separation at both of them. We have compared the photovoltaic properties of the perovskite in p-i-n heterojunctions and also in p-i and i-n junctions. From current-voltage characteristics and impedance spectroscopy, we have observed that two depletion regions were formed at the p-i and i-n interfaces of a p-i-n heterojunction. The two depletion regions operative at p-i-n heterojunctions have yielded better photovoltaic properties as compared to devices having one depletion region in the p-i or the i-n junction. The results evidenced photovoltaic devices based on all-inorganic oxide, nontoxic, and perovskite materials.


A Chemical Approach for Profiling Intracellular AKT Signaling Dynamics from Single Cells.
We present here a novel chemical method to continuously analyze intracellular AKT signaling activities at single-cell resolution, without genetic manipulations. A pair of cyclic peptide-based fluorescent probes were developed to recognize the phosphorylated Ser474 site and a distal epitope on AKT. A Förster resonance energy transfer signal is generated upon concurrent binding of the two probes onto the same AKT protein, which is contingent upon the Ser474 phosphorylation. Intracellular delivery of the probes enabled dynamic measurements of the AKT signaling activities. We further implemented this detection strategy on a microwell single-cell platform, and interrogated the AKT signaling dynamics in a human glioblastoma cell line. We resolved unique features of the single-cell signaling dynamics following different perturbations. Our study provided the first example of monitoring the temporal evolution of cellular signaling heterogeneities and unveiled biological information that was inaccessible to other methods.


Examining visual complexity and its influence on perceived duration.
We investigated whether visual complexity of novel abstract patterns affects perceived duration. Previous research has reported that complex visual stimuli led to an underestimation of durations. However, to clarify the nature of the time estimation process, it is necessary to establish which component of image complexity, spatial or semantic, plays the critical role. Here we tested the impact of specific spatial properties. We used unfamiliar and abstract patterns made using black-and-white checkerboards in which the difference between stimuli was exclusively in configuration. Visual complexity was quantified by the GIF index based on a compression algorithm, which scanned the pattern in both horizontal and vertical directions. This metric correlated positively with subjective complexity (Experiment 1A). In the second study, we increased variability in the stimuli by changing the number of items across patterns while keeping overall size constant. A high positive correlation was found between objective and subjective complexity (r = 0.95) (Experiment 2A). In Experiments 1B and 2B, observers estimated pattern durations in seconds using a continuous scale. A multilevel linear analysis found that perceived duration was not predicted by visual complexity for either of the two sets of stimuli. These results provide new constraints to theories of time perception, hypothesizing that complexity leads to an underestimation of duration when it reduces attention to time.