Search

Photon-counting detector (PCD)-based computed tomography (CT) offers several advantages over conventional energy-integrating detector-based CT. Among them, the ability to discriminate energy exhibits significant potential for clinical applications because it provides material-specific information. That is, material decomposition (MD) can be achieved through energy discrimination. In this study, deep learning-based material decomposition was performed using live animal data. We propose MD-Unet, which is a deep learning strategy for material decomposition based on an Unet architecture trained with data from three energy bins. To mitigate the data insufficiency, we developed a pretrained model incorporating various simulation data forms and augmentation strategies. Incorporating these approaches into model training results in enhanced precision in material decomposition, thereby enabling the identification of distinct materials at individual pixel locations. The trained network was applied to the acquired animal data to evaluate material decomposition results. Compared with conventional methods, the newly generated MD-Unet demonstrated more accurate material decomposition imaging. Moreover, the network demonstrated an improved material decomposition ability and significantly reduced noise. In addition, they can potentially offer an enhancement level similar to that of a typical contrast agent. This implies that it can acquire images of the same quality with fewer contrast agents administered to patients, thereby demonstrating its significant clinical value., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Han et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Chronic pain is a substantial health problem with a high prevalence of comorbid depression. To understand the link between chronic pain and depression, cognitive factors including pain catastrophising and pain self-efficacy have been theorised as significant contributing variables. There is relatively strong evidence that pain catastrophising mediates the relationship between pain severity and depression symptoms. There is also emerging evidence that the mediation role of pain catastrophising may vary as a function of pain self-efficacy. However, it is unknown whether this model will apply in a tertiary pain clinic sample. Thus, this study aimed to examine the respective moderating and mediating roles of pain self-efficacy and pain catastrophising on the association between pain severity and depressive symptoms in a large clinical sample of Australian adults living with chronic pain. Participants (n = 1195) completed all questionnaire measures prior to their first appointments at one tertiary pain service. As expected, the PROCESS path analysis showed that pain catastrophising mediated the relationship between pain severity and depressive symptoms. Further, there was support for the moderating effect of pain self-efficacy; as pain self-efficacy decreased, the relationship strengthened between both pain severity and pain catastrophising, as well as pain catastrophising and depressive symptoms. These findings may have important clinical implications including how relationships between these factors may be considered in the provision of care for those with chronic pain. Notably, these measures could be used in triaging processes to inform treatment decisions., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Kardash et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Skill competency assessments induce stress and anxiety and may affect nursing student performance. Little is known about stress and perceived anxiety levels and their relationship in the mock skill competency assessment., Methods: A cross-sectional study was conducted to examine the stress levels (as assessed by heart rate variability, HRV) and perceived anxiety before, during and after the mock skill competency assessment, and to explore their relationships to performance in a total of ninety first-year undergraduate nursing students., Results: The HRV decreased significantly during the assessment and increased significantly 10 min after the assessment (p < 0.01). Higher performers showed significantly lower HRV during and after the assessment (p < 0.01). The assessment score was negatively correlated with HRV during and after the assessment (p < 0.05)., Conclusions: Considering assessment-related stress and anxiety through a mock assessment prior to the actual skill assessment provides implications for future nursing education., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Wong et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Aim: The aim of this literature review was to identify, summarize, and critically appraise available empirical articles on the knowledge, attitudes, and practices towards childhood fever management among South-East and East Asian parents., Design: A literature review following PRISMA., Methods: Articles were limited to those available in the English language. Articles had to be empirical studies that used a qualitative or quantitative research design with full-text available; focus on parental knowledge, attitudes, and practices towards fever; and be published in South-East and East Asia. Searches were conducted with CINAHL, PubMed and Scopus from inception to June 2022, and eleven articles were included after removing duplicates and excluding irrelevant articles., Results: Narrative synthesis was conducted according to four themes: source of fever information, knowledge level, attitudes, and practices towards childhood fever. Parents showed different fever knowledge needs and various information-seeking behaviors. A low level of fever knowledge was revealed in terms of temperature, fever causes, potential harms and influencing factors. South-East and East Asian parents mainly reported anxiety, concerns and fever phobia. Fever assessment methods and fever management strategies varied based on parents' cultural background and beliefs., Conclusions: The findings of this review highlight that inadequacy of fever knowledge and negative attitudes towards childhood fever exist in South-East and East Asian parents. Parents have diverse cultural practices during their children's febrile episodes. However, some of them conflict with current medical guidelines, as they prioritize fever and body temperature reduction. This raises questions about their effectiveness and safety. Although some of them are medically discouraged, there are others that have been proven beneficial for the symptomatic relief of childhood fever. The results indicate an urgent need to develop a cultural-sensitive educational intervention for childhood fever management among South-East and East Asian parents. Unified educational interventions are needed to address parental concerns and fever-related knowledge needs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Ng et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Cognitive flexibility (CF) enables individuals to readily shift from one concept or mode of practice/thoughts to another in response to changes in the environment and feedback, making CF vital to optimise success in obtaining goals. However, how CF relates to other executive functions (e.g., working memory, response inhibition), mental abilities (e.g., creativity, literacy, numeracy, intelligence, structure learning), and social factors (e.g., multilingualism, tolerance of uncertainty, perceived social support, social decision-making) is less well understood. The current study aims to (1) establish the construct validity of CF in relation to other executive function skills and intelligence, and (2) elucidate specific relationships between CF, structure learning, creativity, career decision making and planning, and other life skills., Methods: This study will recruit up to 400 healthy Singaporean young adults (age 18-30) to complete a wide range of cognitive tasks and social questionnaires/tasks. The richness of the task/questionnaire battery and within-participant administration enables us to use computational modelling and structural equation modelling to examine connections between the latent constructs of interest., Significance and Impact: The current study is the first systematic investigation into the construct validity of CF and its interrelationship with other important cognitive skills such as learning and creativity, within an Asian context. The study will further explore the concept of CF as a non-unitary construct, a novel theoretical proposition in the field. The inclusion of a structure learning paradigm is intended to inform future development of a novel intervention paradigm to enhance CF. Finally, the results of the study will be useful for informing classroom pedagogy and the design of lifelong learning policies and curricula, as part of the wider remit of the Cambridge-NTU Centre for Lifelong Learning and Individualised Cognition (CLIC)., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Background: This is a pilot proof-of-concept study to evaluate the utility of a custom 15-gene circulating tumor DNA (ctDNA) panel as a potential companion molecular next-generation sequencing (NGS) assay for identifying somatic single nucleotide variants and indels in non-small-cell lung cancer (NSCLC) patients. The custom panel covers the hotspot mutations in EGFR, KRAS, NRAS, BRAF, PIK3CA, ERBB2, MET, KIT, PDGFRA, ALK, ROS1, RET, NTRK1, NTRK2 and NTRK3 genes which serve as biomarkers for guiding treatment decisions in NSCLC patients., Method: The custom 15-gene ctDNA NGS panel was designed using ArcherDX Assay Designer. A total of 20 ng or 50 ng input ctDNA was used to construct the libraries. The analytical performance was evaluated using reference standards at different allellic frequencies (0.1%, 1%, 5% and parental). The clinical performance was evaluated using plasma samples collected from 10 treatment naïve advanced stage III or IV NSCLC patients who were tested for tissue EGFR mutations. The bioinformatics analysis was performed using the proprietary Archer Analysis Software., Results: For the analytical validation, we achieved 100% sensitivity and specificity for the detection of known mutations in the reference standards. The limit of detection was 1% allelic frequency. Clinical validation showed that the clinical sensitivity and specificity of the assay for detecting EGFR mutation were 83.3% and 100% respectively. In addition, the NGS panel also detected other mutations of uncertain significance in 6 out of 10 patients., Conclusion: This preliminary analysis showed that the custom 15-gene ctDNA NGS panel demonstrated good analytical and clinical performances for the EGFR mutation. Further studies incorporating the validation of other candidate gene mutations are warranted., Competing Interests: The authors declare that they have no conflict of interest.

Plant-based anticancer agents have the potential to stimulate the immune system to act against cancer cells. A standardized bioactive subfraction of the Malaysian herb, Strobilanthes crispus (L.) Blume (S. crispus) termed F3, demonstrates strong anticancer effects in both in vitro and in vivo models. The anticancer effects might be attributable to its immunomodulatory properties as S. crispus has been traditionally used to enhance the immune system. The current study examined whether F3 could stimulate anti-tumorigenic immunogenicity against 4T1 cells in vitro and in 4T1 cell-induced mammary carcinoma mouse model. We observed that F3 induced significant increase in MHC class I and class II molecules. CD4+, CD8+ and IL-2+ (p<0.05 for all) cells infiltration was also significantly increased in the breast tumor microenvironment of F3-treated mice compared with the tumors of untreated mice. The number of CD68+ macrophages was significantly lower in F3-treated mice. We conclude that the antitumor and antimetastatic effects of S. crispus involve strong infiltration of T cells in breast cancer potentially through increased tumor antigen presentation via MHC proteins, as well as reduction of infiltrating tumor-associated macrophages., Competing Interests: The authors have declared that no competing interests exist.

There remains a limited understanding of the HIV prevention and treatment needs among female sex workers in many parts of the world. Systematic reviews of existing literature can help fill this gap; however, well-done systematic reviews are time-demanding and labor-intensive. Here, we propose an automatic document classification approach to a systematic review to significantly reduce the effort in reviewing documents and optimizing empiric decision making. We first describe a manual document classification procedure that is used to curate a pertinent training dataset and then propose three classifiers: a keyword-guided method, a cluster analysis-based method, and a random forest approach that utilizes a large set of feature tokens. This approach is used to identify documents studying female sex workers that contain content relevant to either HIV or experienced violence. We compare the performance of the three classifiers by cross-validation in terms of area under the curve of the receiver operating characteristic and precision and recall plot, and found random forest approach reduces the amount of manual reading for our example by 80%; in sensitivity analysis, we found that even trained with only 10% of data, the classifier can still avoid reading 75% of future documents (68% of total) while retaining 80% of relevant documents. In sum, the automated procedure of document classification presented here could improve both the precision and efficiency of systematic reviews and facilitate live reviews, where reviews are updated regularly. We expect to obtain a reasonable classifier by taking 20% of retrieved documents as training samples. The proposed classifier could also be used for more meaningfully assembling literature in other research areas and for rapid documents screening with a tight schedule, such as COVID-related work during the crisis., Competing Interests: The authors have declared that no competing interests exist.

Objectives: While depression and anxiety are common in women and men of reproductive age, preconception interventions to optimize the health of individuals with mental illness before pregnancy is limited and focuses primarily on psychotropic medication management. Comparing individuals with depression, anxiety, and comorbidity to those with neither condition, we identified areas of preconception care optimization related to psychosocial risk factors, general physical health, medication use, and uptake of high-risk health behaviours. We also investigated differences in preconception health care use, attitudes, and knowledge., Method: We conducted a nationwide survey of 621 women (n = 529) and men (n = 92) across Canada who were planning a pregnancy within five years, including those with lifetime or current depression (n = 38), anxiety (n = 55), and comorbidity (n = 104) and those without mental illness (n = 413). Individuals with depression, anxiety, and comorbidity were compared to individuals without mental illness using logistic regression, adjusted for age, sex, and education level., Results: Individuals with a lifetime or current mental illness were significantly more likely to have several risk factors for suboptimal reproductive and perinatal outcomes, including increased rates of obesity, stress, fatigue, loneliness, number of chronic health conditions, and medication use. Further, they were more likely to have high-risk health behaviours including increased substance use, internet addiction, poorer eating habits, and decreased physical activity. By assessing depression, anxiety, or both separately, we also determined there was variation in risk factors by mental illness type., Conclusion: Our nationwide study is one of the first and largest to examine the preconception care needs of women and men with a lifetime or current mental illness who are pregnancy-planning. We found this population has many important reproductive and perinatal risk factors that are modifiable via preconception interventions which could have a significant positive impact on their health trajectories and those of their future children., Competing Interests: The authors have no conflicts of interest to declare.

Objectives: To establish the factorial structure and internal consistency of the Internet Addiction Test (IAT) in parents, the level and correlates of problematic internet use, and patterns and types of screen use., Study Design: Data were collected through an online questionnaire about preconception health among Canadian women and men with ≥1 child. The questionnaire included the IAT and questions about time spent on screens by device type, use of screens during meals and in the bedroom, and perceptions of overuse. Factor analysis was completed to determine the factorial structure of the IAT, with multivariable linear regression used to determine correlates of the IAT., Results: The sample included 1,156 respondents (mean age: 34.3 years; 83.1% female). The IAT had two factors: "impairment in time management" and "impairment in socio-emotional functioning" of which respondents had more impairment in time management than socio-emotional functioning. Based on the original IAT, 19.4% of respondents would be classified as having a mild internet use problem with 3.0% having a moderate or severe issue. In the multivariable model, perceived stress (b = .28, SE = .05, p < .001) and depressive symptoms (b = .24, SE = .10, p = .017) were associated with higher IAT scores. Handheld mobile devices were the most common type of screen used (mean = 3 hours/day) followed by watching television (mean = 2 hours/day)., Conclusion: Parents spent a significant portion of their time each day using screens, particularly handheld mobile devices. The disruption caused by mobile devices may hinder opportunities for positive parent-child interactions, demonstrating the need for resources to support parents ever-growing use of technologies., Competing Interests: The authors have declared that no competing interests exist.

Background: Anxiety, nausea and vomiting are common side effects suffered by paediatric patients receiving chemotherapy. Emerging evidence supports the efficacy of immersive virtual reality (IVR) on improving anxiety and distress symptoms including nausea and vomiting among this vulnerable group. This trial aims to assess the feasibility and acceptability of IVR for preventing and managing anxiety, nausea and vomiting among paediatric cancer patients receiving their first chemotherapy., Method and Analysis: An exploratory trial supplemented by qualitative methods will be conducted. We will recruit 20 paediatric patients who are aged between 6 and 12 years, chemotherapy naïve, scheduled to receive their first intravenous chemotherapy and able to understand Chinese. Participants will be randomly allocated to intervention or control groups. The intervention group will receive the IVR intervention for three sessions as follows: 4 hours before chemotherapy, 5 minutes before and during their first course chemotherapy and 5 minutes before and during their second course chemotherapy. The control group will receive standard care only. Main outcome measures included (1) key parameters for the design of a definitive trial (i.e. screening, eligibility, consent and withdrawal rates); (2) anxiety, anticipatory and acute chemotherapy-induced nausea and vomiting for collection of preliminary data; (3) feasibility and acceptability of the intervention. Semi-structured interviews will be conducted with patients, parents and oncology nurses. Generalized estimating equations model will be used to compare each of the outcome measures across the time points between the two groups. Qualitative data will be analysed by conventional content analysis., Expected Results: The results of this exploratory trial will inform the design and conduct of future definitive trial., Trial Registration Number: ChiCTR1900021694; Pre-results., Competing Interests: The authors have declared that no competing interests exist.

Encouraging citizens to use health checkup services is a health promotion strategy. In nations with aging populations, ensuring equitable use of health check-ups by senior citizens is a public health concern. The objective of this research was to quantify income-related inequality and its effect on the use of health checkup services in Taiwan during the 2007-2008 global financial crisis. We used the 2005 and 2009 datasets of the Taiwan National Health Interview Surveys to assess how income-related inequality influenced health check-up use among older adults in Taiwan during the 2007-2008 financial crisis. Corrected concentration indices (CCIs) were calculated and decomposed to determine the influences of explanatory variables. The dependent variable was whether participants had used free senior health check-ups in the past year, and the determinant factors were health behavior, health situation, socioeconomic and demographic factors, and area health care resources accessibility factors. The study assessed 2,460 older adults from the 2005 dataset and 2,514 such individuals from the 2009 dataset. The utilization of health check-ups increased from 21.6% in 2005 to 34.0% in 2009. Income-related inequality in the use of health check-up services was generally tilted toward the higher income individuals among both women and men in 2005 and 2009, and income-related inequality decreased among women group and increased among men group with non significantly from 2005 to 2009 (women: CCI decreased from.0738 in 2005 to.0658 in 2009; men: CCI increased from.1068 in 2005 to.1256 in 2009). We analyzed the effect of explanatory factors on men's and women's intention to use health check-ups by using a probit model. After controlling for other factors, we determined that income significantly influenced women's health check-up service use in 2005 and men's in 2005 and 2009. Positive health behavior significantly increased health check-up services use among men and women group after the financial crisis, and negative health behavior significantly reduced health check-ups use among men across financial crisis. The 2008 global financial crisis strengthened the effect on health check-ups use of income-related inequality of elderly men, especially in older adults with negative health behaviors. Elderly men with negative health behaviors tended to contribute more income-related inequality in use health check-up services after the financial crisis. Health promotion initiatives should focus their efforts on elderly men with negative health behaviors., Competing Interests: The authors have declared that no competing interests exist.

Metal artifacts are considered a major challenge in computed tomography (CT) as these adversely affect the diagnosis and treatment of patients. Several approaches have been developed to address this problem. The present study explored the clinical potential of a novel photon-counting detector (PCD) CT system in reducing metal artifacts in head CT scans. In particular, we studied the recovery of an oral tumor region located under metal artifacts after correction. Three energy thresholds were used to group data into three bins (bin 1: low-energy, bin 2: middle-energy, and bin 3: high-energy) in the prototype PCD CT system. Three types of physical phantoms were scanned on the prototype PCD CT system. First, we assessed the accuracy of iodine quantification using iodine phantoms at varying concentrations. Second, we evaluated the performance of material decomposition (MD) and virtual monochromatic images (VMIs) using a multi-energy CT phantom. Third, we designed an ATOM phantom with metal insertions to verify the effect of the proposed metal artifact reduction. In particular, we placed an insertion-mimicking an iodine-enhanced oral tumor in the beam path of metallic objects. Normalized metal artifact reduction (NMAR) was performed for each energy bin image, followed by an image-based MD and VMI reconstruction. Image quality was analyzed quantitatively by contrast-to-noise ratio (CNR) measurements. The results of iodine quantification showed a good match between the true and measured iodine concentrations. Furthermore, as expected, the contrast between iodine and the surrounding material was higher in bin 1 image than in bin 3 image. On the other hand, the bin 3 image of the ATOM phantom showed fewer metal artifacts than the bin 1 image because of the higher photon energy. The result of quantitative assessment demonstrated that the 40-keV VMI (CNR: 20.6 ± 1.2) with NMAR and MD remarkably increased the contrast of the iodine-enhanced region compared with that of the conventional images (CNR: 10.4 ± 0.5) having 30 to 140 keV energy levels. The PCD-based multi-energy CT imaging has immense potential to maximize the contrast of the target tissue and reduce metal artifacts simultaneously. We believe that it would open the door to novel applications for the diagnosis and treatment of several diseases., Competing Interests: The authors have read the journal’s policy and have the following competing interests: All authors are paid employees of Samsung Electronics Co., Ltd. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

The Southwest Pacific represents an independent biogeographic province for deep-sea hydrothermal vent fauna. Different degrees of genetic connectivity among vent fields in Manus, North Fiji and Lau Basins have been reported for various molluscan and crustacean species, presumably reflecting their different levels of dispersal ability as swimming larvae. The present study investigates the population connectivity of the hydrothermal vent limpet Shinkailepas tollmanni (family Phenacolepadidae) in the Southwest Pacific. Our analyses using mitochondrial COI-gene sequences and shell morphometric traits suggest a panmictic population structure throughout its geographic and bathymetric ranges, spanning 4,000 km from the westernmost Manus Basin (151ºE; 1,300 m deep) to the easternmost Lau Basin (176ºE; 2,720 m). The measurements of its embryonic and larval shells demonstrate that the species hatches as a planktotrophic veliger larva with an embryonic shell diameter of 170-180 μm and settles at the vent environment with the larval shell diameter of 750-770 μm. This substantial growth as a feeding larva, ca. 80 times in volume, is comparable or even greater than those of confamilial species in the hydrothermal-vent and methane-seep environments in the Northwest Pacific and Atlantic Oceans. Large pigmented eyes in newly settled juveniles are another common feature in this and other phenacolepadids inhabiting the chemosynthetic environments. These results put together suggest that the larvae of S. tollmanni migrate vertically from deep-sea vents to surface waters to take advantages of richer food supplies and faster currents and stay pelagic for an extended period of time (> 1 year), as previously indicated for the confamilial species., Competing Interests: The authors have read the journal’s policy and have the following potential competing interests: ADT is the CEO of Blackbeard Biologic: Science and Environmental Advisors. This company did not yet exist when ADT was involved in working on this study, and therefore did not provide any funding in the form of salary. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare.

Purpose: To determine whether SRD5A2 promoter methylation is associated with cancer progression during androgen deprivation therapy (ADT) in CRPC., Patients and Methods: In a Local CRPC cohort, 42 prostatic specimens were collected from patients who were diagnosed as CRPC and underwent transurethral resection of the prostate (TURP) at Massachusetts General Hospital (MGH). In a metastatic CRPC (Met CRPC) cohort, 12 metastatic biopsies were collected from CRPC patients who would be treated with abiraterone plus dutasteride (Clinical Trial NCT01393730). As controls, 36 benign prostatic specimens were collected from patients undergoing prostate reduction surgery for symptoms of bladder outlet obstruction secondary to benign prostatic hyperplasia (BPH). The methylation status of cytosine-phosphate-guanine (CpG) site(s) at SRD5A2 promoter regions was tested., Results: Compared with benign prostatic tissue, CRPC samples demonstrated higher SRD5A2 methylation in the whole promoter region (Local CRPC cohort: P < 0.001; Met CRPC cohort: P <0.05). In Local CRPC cohort, a higher ratio of methylation was correlated with better OS (R2 = 0.33, P = 0.013). Hypermethylation of specific regions (nucleotides -434 to -4 [CpG# -39 to CpG# -2]) was associated with a better OS (11.3±5.8 vs 6.4±4.4 years, P = 0.001) and PFS (8.4±5.4 vs 4.5±3.9 years, P = 0.003) with cutoff value of 37.9%. Multivariate analysis showed that SRD5A2 methylation was associated with OS independently (whole promoter region: P = 0.035; specific region: P = 0.02)., Conclusion: Our study demonstrate that SRD5A2 methylation in promoter regions, specifically at CpG# -39 to -2, is significantly associated with better survival for CRPC patients treated with ADT. Recognition of epigenetic modifications of SRD5A2 may affect the choices and sequence of available therapies for management of CRPC., Competing Interests: The authors have declared that no competing interests exist.

The maturation of photon-counting detector (PCD) technology promises to enhance routine CT imaging applications with high-fidelity spectral information. In this paper, we demonstrate the power of this synergy and our complementary reconstruction techniques, performing 4D, cardiac PCD-CT data acquisition and reconstruction in a mouse model of atherosclerosis, including calcified plaque. Specifically, in vivo cardiac micro-CT scans were performed in four ApoE knockout mice, following their development of calcified plaques. The scans were performed with a prototype PCD (DECTRIS, Ltd.) with 4 energy thresholds. Projections were sampled every 10 ms with a 10 ms exposure, allowing the reconstruction of 10 cardiac phases at each of 4 energies (40 total 3D volumes per mouse scan). Reconstruction was performed iteratively using the split Bregman method with constraints on spectral rank and spatio-temporal gradient sparsity. The reconstructed images represent the first in vivo, 4D PCD-CT data in a mouse model of atherosclerosis. Robust regularization during iterative reconstruction yields high-fidelity results: an 8-fold reduction in noise standard deviation for the highest energy threshold (relative to unregularized algebraic reconstruction), while absolute spectral bias measurements remain below 13 Hounsfield units across all energy thresholds and scans. Qualitatively, image domain material decomposition results show clear separation of iodinated contrast and soft tissue from calcified plaque in the in vivo data. Quantitatively, spatial, spectral, and temporal fidelity are verified through a water phantom scan and a realistic MOBY phantom simulation experiment: spatial resolution is robustly preserved by iterative reconstruction (10% MTF: 2.8-3.0 lp/mm), left-ventricle, cardiac functional metrics can be measured from iodine map segmentations with ~1% error, and small calcifications (615 μm) can be detected during slow moving phases of the cardiac cycle. Given these preliminary results, we believe that PCD technology will enhance dynamic CT imaging applications with high-fidelity spectral and material information., Competing Interests: We have the following interests: This work was made possible by the loan of a prototype SANTIS 0804 ME photon-counting x-ray detector from DECTRIS, Ltd. (Baden-Dättwil, Switzerland; https://www.dectris.com/). There are no patents, products in development, or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.

To understand the origin, migration, and distribution of organisms across disjunct deep-sea vent habitats, previous studies have documented the population genetic structures of widely distributed fauna, such as gastropods, bivalves, barnacles, and squat lobsters. However, a limited number of investigations has been conducted in the Southwest Pacific Ocean, and many questions remain. In this study, we determined the population structure of the bythograeid crab Austinograea alayseae from three adjacent vent systems (Manus Basin, North Fiji Basin, and Tonga Arc) in the Southwest Pacific Ocean using the sequences of two mitochondrial genes (COI and 16S rDNA) and one nuclear gene (28S rDNA). Populations were divided into a Manus clade and a North Fiji-Tonga clade, with sequence divergence values in the middle of the barcoding gap for bythograeids. We inferred that hydrographic and/or physical barriers act on the gene flow of A. alayseae between the Manus and North Fiji basins. Austinograea alayseae individuals interact freely between the North Fiji Basin and the Lau Basin (Tonga Arc). Although further studies of genetic differentiation over a geological time scale, life-history attributes, and genome-based population genetics are needed to improve our understanding of the evolutionary history of A. alayseae, our results contribute to elucidating the phylogeny, evolution, and biogeography of bythograeids., Competing Interests: The authors have declared that no competing interests exist.

Background: Hand grip strength (HGS) is associated with a number of causes resulting in cardiovascular death, in addition to bone fragility, and the presence of sarcopenia. The goal of our study was to analyze HGS of students based on chronological and biological age and propose normative standards for children and adolescents from Chile., Methods: We studied 4604 school children of both sexes between the ages of 6.0 and 17.9 years of age. Weight, standing height, sitting height, and hand grip strength (HGS- right and left) were measured. The Body Mass Index (BMI) was calculated, and the biological age was calculated by using age at peak height velocity (APHV)., Results: When arranged by chronological age, no significant differences occurred in HGS between both sexes of school children from age 6 to 12 years of age. However, from ages 13 to 17, males showed greater HGS than females. Significant differences also emerged between both sexes and at all levels for biological age (APHV). For males, chronological age explained the HGS occurring between 0.74 to 0.75% and for females between 0.54 to 0.59%. For males, biological age explained the HGS for the range of 0.79 to 0.80% and 0.62 to 0.67% for females. The normative data for HGS for both sexes is expressed in percentiles., Conclusions: HGS during childhood and adolescence needs be analyzed and interpreted in terms of biological age rather than chronological age. The normative data to evaluate the HGS are a tool that can help professionals working in clinical and epidemiological contexts., Competing Interests: The authors have declared that no competing interests exist.

Background: Electron microscopy (EM) characteristics of the urothelium in interstitial cystitis/bladder pain syndrome (IC/BPS) and their association with clinical condition are unclear., Methods: Ten IC/BPS patients who were admitted for hydrodistention and 5 patients with stress urinary incontinence (control patients) were enrolled. All patients provided detailed clinical histories and underwent urodynamic studies. Cystoscopic bladder biopsies were obtained and processed for transmission EM (TEM) and scanning EM (SEM). The severity of the urothelium findings was graded on a 4-point scale (0: none, 1: mild, 2: moderate, and 3: severe). The EM findings between IC/BPS and control patients were compared; the results were analyzed using the chi-square test., Results: Compared with the urothelium of control patients, the urothelium of IC/BPS patients had more severe defects of the urothelial cell layers and integrity of umbrella cells in TEM (p = 0.045 and 0.01, respectively). In SEM, umbrella cell pleomorphism increased and microplicae of the cell membrane decreased in the IC/BPS group, and both were more severe than in the control group (p = 0.022 and 0.007, respectively). The patients with moderate to severe defects of umbrella cell integrity had more severe bladder pain and smaller maximal bladder capacity (MBC) (both p = 0.010). Patients with moderate to severe defects in microplicae of the cell membrane had smaller cystometric bladder capacity and MBC (p = 0.037 and 0.047, respectively)., Conclusions: The results revealed significant urothelium defects in IC/BPS, especially in the umbrella cells. Defects of umbrella cells may play an important role in the pathogenesis of IC/BPS., Competing Interests: The authors have declared that no competing interests exist.

Various sociocultural factors affect healthcare-seeking behaviors. In Taiwanese society, superstitions and lunar festivals play important roles in people's lives. We investigated the impact of "Ghost Month" (the 7th lunar month) and Chinese New Year (the 12th lunar month and the 1st lunar month of the following year) on the number of elective surgeries and emergent surgeries in Taiwan. The number of total knee replacement (TKR) surgeries and proximal femur fracture (PFF) surgeries in each lunar month from 2000 to 2011 were extracted from the Taiwan National Health Insurance Database, a computerized and population-based database. Patients were then sorted by location of residence or gender. The average number of TKR surgeries performed was significantly lower during the 1st, 7th, and 12th lunar months in urban areas, whereas in rural areas this trend was only evident in the 7th and 12th lunar months. There was however, no significant difference in the average number of PFF surgeries in each lunar month except for an increase seen in the 1st lunar month in rural patients (p<0.05). When sorted by gender, the average number of TKR surgeries was significantly decreased in the 7th and 12th lunar months in male patients, and decreased in the 1st, 7th, and 12th lunar months in female patients. In contrast, there was no difference in the average numbers of PFF surgeries in the 7th and 12th lunar months either in male or female patients. We proposed that the timing of elective surgeries such as TKR might be influenced by Ghost Month and Chinese New Year; however, emergent PFF surgeries were not significantly influenced by sociocultural beliefs and taboos in Taiwan.

Background: Increased incidence of intraoperative awareness was reported in critically ill patients during major operations, particularly under total intravenous (TIVA) or endotracheal general anesthesia (ETGA). However, the incidence and effect of anesthesia techniques on awareness in generally healthy, non-critically ill patients during operations have yet to receive significant attention., Methods and Results: This retrospective matched case-control study was conducted between January 2009 to December 2014. Surgical patients (ASA physical status I-III) whom reported intraoperative awareness during this study period were interviewed and their medical records were reviewed. The potential risk factors for awareness were compared with the non-case matched controls, who were randomly selected from the database. A total of 61436 patients were included and 16 definite cases of intraoperative awareness were identified. Patients who received ETGA and TIVA had significantly higher incidence of developing awareness compared to those who were anesthetized using laryngeal masks (LMA) (P = 0.03). Compared with the matched controls (n = 80), longer anesthesia time was associated with increased incidence of awareness (odds ratio 2.04; 95% CI 1.30-3.20, per hour increase). Perioperative use of muscle relaxant was also associated with increased incidence of awareness, while significantly lower incidence of awareness was found in patients who were anesthetized with volatile anesthetics., Conclusions: The overall incidence of awareness was 0.023% in the ASA≤ III surgical patients who received general anesthesia. Anesthesia with a laryngeal mask under spontaneous ventilation and supplemented with volatile anesthetics may be the preferred anesthesia technique in generally healthy patients in order to provide a lower risk of intraoperative awareness.

Background: The Dual Energy X-Ray Absorptiometry (DXA) is the gold standard for measuring BMD and bone mineral content (BMC). In general, DXA is ideal for pediatric use. However, the development of specific standards for particular geographic regions limits its use and application for certain socio-cultural contexts. Additionally, the anthropometry may be a low cost and easy to use alternative method in epidemiological contexts. The goal of our study was to develop regression equations for predicting bone health of children and adolescents based on anthropometric indicators to propose reference values based on age and sex., Methods: 3020 students (1567 males and 1453 females) ranging in ages 4.0 to 18.9 were studied from the Maule Region (Chile). Anthropometric variables evaluated included: weight, standing height, sitting height, forearm length, and femur diameter. A total body scan (without the head) was conducted by means of the Dual Energy X-Ray Absorptiometry. Bone mineral density (BMD) and the bone mineral content (BMC) were also determined. Calcium consumption was controlled for by recording the intake of the three last days prior to the evaluation. Body Mass Index (BMI) was calculated, and somatic maturation was determined by using the years of peak growth rate (APHV)., Results: Four regression models were generated to calculate bone health: for males BMD = (R2 = 0.79) and BMC = (R2 = 0.84) and for the females BMD = (R2 = 0.76) and BMC = (R2 = 0.83). Percentiles were developed by using the LMS method (p3, p5, p15, p25, p50, p75, p85, p95 and p97)., Conclusions: Regression equations and reference curves were developed to assess the bone health of Chilean children and adolescents. These instruments help identify children with potential underlying problems in bone mineralization during the growth stage and biological maturation.

Introduction: A high hemoglobin glycation index (HGI) and glycated hemoglobin (HbA1c) level are associated with greater inflammatory status, and dipeptidyl peptidase-4 (DPP-4) inhibitors can suppress inflammation. We aimed to evaluate the relationship between HGI and the therapeutic effect of DPP-4 inhibitors., Methods: This retrospective cohort study followed 468 patients with type 2 diabetes receiving DPP-4 inhibitor treatment for 1 year. Estimated HbA1c was calculated using a linear regression equation derived from another 2969 randomly extracted patients with type 2 diabetes based on fasting plasma glucose (FPG) level. The subjects were divided into two groups based on HGI (HGI = observed HbA1c - estimated HbA1c). Mixed model repeated measures were used to compare the treatment efficacy after 1 year in patients with a low (HGI<0, n = 199) and high HGI (HGI≧0, n = 269)., Results: There were no significant group differences in mean changes of FPG after 1 year (-12.8 and -13.4 mg/dL in the low and high HGI groups, respectively). However, the patients with a high HGI had a significantly greater reduction in HbA1c from baseline compared to those with a low HGI (-1.9 versus -0.3% [-20.8 versus -3.3 mmol/mol]). Improvements in glycemic control were statistically significantly associated with the tested DPP-4 inhibitors in the high HGI group (-2.4, -1.4, -1.2 and -2.2% [-26.2, -15.3, -13.1 and -24.0 mmol/mol] for vildagliptin, linagliptin, saxagliptin and sitagliptin, respectively) but not in the low HGI group., Conclusions: The HGI index derived from FPG and HbA1c may be able to identify who will have a better response to DPP-4 inhibitors., Competing Interests: The authors have declared that no competing interests exist.

Antrodia camphorata is a fungus that is endemic to Taiwan, and its fruiting body has been used as a folk medicine for the prevention or treatment of diverse diseases. The present study is aimed at investigating the antimelanogenesis and antioxidation effect of the ethanolic extract of Antrodia camphorata fruiting body (EE-AC), as well as its antiproliferation effects in B16-F0 melanoma cells. Regarding antimelanogenic effects, EE-AC had effective cupric ions reducing capacity and expressed more potent inhibitory effect than kojic acid on mushroom tyrosinase activity. Moreover, EE-AC significantly inhibited cellular tyrosinase activity and the melanin content in B16-F0 cells at 12.5 μg/mL concentration without cell toxicities. Regarding antioxidant effects, EE-AC exhibited potent DPPH radical- and SOD-like-scavenging activities. Regarding antiproliferative effects, EE-AC exhibited a selective cytotoxic effect and markedly inhibited the migration ability of B16-F0 cells. EE-AC increased the population of B16-F0 cells at sub-G1 phase of the cell cycle. EE-AC also caused the increase of early apoptotic cells and chromatin condensation, which indicated the apoptotic effects in B16-F0 cells. We demonstrated that EE-AC possessed antimelanogenic, antioxidant and anti-skin cancer actions. The results would contribute to the development and application of cosmetics, healthy food and pharmaceuticals., Competing Interests: The authors have declared that no competing interests exist.

Visual crowding-the inability to see an object when it is surrounded by flankers in the periphery-does not block semantic activation: unrecognizable words due to visual crowding still generated robust semantic priming in subsequent lexical decision tasks. Based on the previous finding, the current study further explored whether unrecognizable crowded words can be temporally integrated into a phrase. By showing one word at a time, we presented Chinese four-word idioms with either a congruent or incongruent ending word in order to examine whether the three preceding crowded words can be temporally integrated to form a semantic context so as to affect the processing of the ending word. Results from both behavioral (Experiment 1) and Event-Related Potential (Experiment 2 and 3) measures showed congruency effect in only the non-crowded condition, which does not support the existence of unconscious multi-word integration. Aside from four-word idioms, we also found that two-word (modifier + adjective combination) integration-the simplest kind of temporal semantic integration-did not occur in visual crowding (Experiment 4). Our findings suggest that integration of temporally separated words might require conscious awareness, at least under the timing conditions tested in the current study.

Unlabelled: Coccydynia is pain in the coccygeal region, and usually treated conservatively. Extracorporeal shock wave therapy (ESWT) was incorporated as non-invasive treatment of many musculoskeletal conditions. However, the effects of ESWT on coccydynia are less discussed. The purpose of this study is to evaluate the effects of ESWT on the outcomes of coccydynia. Patients were allocated to ESWT (n = 20) or physical modality (SIT) group (n = 21) randomly, and received total treatment duration of 4 weeks. The visual analog scale (VAS), Oswestry disability index (ODI), and self-reported satisfaction score were used to assess treatment effects. The VAS and ODI scores were significantly decreased after treatment in both groups, and the decrease in the VAS score was significantly greater in the ESWT group. The mean proportional changes in the ODI scores were greater in the ESWT group than in the SIT group, but the between-group difference was not statistically significant. The patients in the ESWT group had significantly higher subjective satisfaction scores than SIT group. We concluded that ESWT is more effective and satisfactory in reducing discomfort and disability caused by coccydynia than the use of physical modalities. Thus, ESWT is recommended as an alternative treatment option for patients with coccydynia., Trial Registration: ClinicalTrials.gov NCT02313324.

The large quantities of data now being transferred via high-speed networks have made deep packet inspection indispensable for security purposes. Scalable and low-cost signature-based network intrusion detection systems have been developed for deep packet inspection for various software platforms. Traditional approaches that only involve central processing units (CPUs) are now considered inadequate in terms of inspection speed. Graphic processing units (GPUs) have superior parallel processing power, but transmission bottlenecks can reduce optimal GPU efficiency. In this paper we describe our proposal for a hybrid CPU/GPU pattern-matching algorithm (HPMA) that divides and distributes the packet-inspecting workload between a CPU and GPU. All packets are initially inspected by the CPU and filtered using a simple pre-filtering algorithm, and packets that might contain malicious content are sent to the GPU for further inspection. Test results indicate that in terms of random payload traffic, the matching speed of our proposed algorithm was 3.4 times and 2.7 times faster than those of the AC-CPU and AC-GPU algorithms, respectively. Further, HPMA achieved higher energy efficiency than the other tested algorithms.

This paper presents a methodology based on multivariate data analysis for characterizing potential source contributions of emerging contaminants (ECs) detected in 26 river water samples across multi-scape regions during dry and wet seasons. Based on this methodology, we unveil an approach toward potential source contributions of ECs, a concept we refer to as the "Pharmaco-signature." Exploratory analysis of data points has been carried out by unsupervised pattern recognition (hierarchical cluster analysis, HCA) and receptor model (principal component analysis-multiple linear regression, PCA-MLR) in an attempt to demonstrate significant source contributions of ECs in different land-use zone. Robust cluster solutions grouped the database according to different EC profiles. PCA-MLR identified that 58.9% of the mean summed ECs were contributed by domestic impact, 9.7% by antibiotics application, and 31.4% by drug abuse. Diclofenac, ibuprofen, codeine, ampicillin, tetracycline, and erythromycin-H2O have significant pollution risk quotients (RQ>1), indicating potentially high risk to aquatic organisms in Taiwan.

Nanoimprinting lithography (NIL) is a manufacturing process that can produce macroscale surface areas with nanoscale features. In this paper, this technique is used to solve three fundamental issues for the application of localized surface plasmonic resonance (LSPR) in practical clinical measurements: assay sensitivity, chip-to-chip variance, and the ability to perform assays in human serum. Using NIL, arrays of 140 nm square features were fabricated on a sensing area of 1.5 mm x 1.5 mm with low cost. The high reproducibility of NIL allowed for the use of a one-chip, one-measurement approach with 12 individually manufactured surfaces with minimal chip-to-chip variations. To better approximate a real world setting, all chips were modified with a biocompatible, multi-component monolayer and inter-chip variability was assessed by measuring a bioanalyte standard (2.5-75 ng/ml) in the presence of a complex biofluid, human serum. In this setting, nanoimprinted LSPR chips were able to provide sufficient characteristics for a 'low-tech' approach to laboratory-based bioanalyte measurement, including: 1) sufficient size to interface with a common laboratory light source and detector without the need for a microscope, 2) high sensitivity in serum with a cardiac troponin limit of detection of 0.55 ng/ml, and 3) very low variability in chip manufacturing to produce a figure of merit (FOM) of 10.5. These findings drive LSPR closer to technical comparability with ELISA-based assays while preserving the unique particularities of a LSPR based sensor, suitability for multiplexing and miniaturization, and point-of-care detections.

Studies of genetic connectivity and population structure in deep-sea chemosynthetic ecosystems often focus on endosymbiont-hosting species that are directly dependent on chemical energy extracted from vent effluent for survival. Relatively little attention has been paid to vent-associated species that are not exclusively dependent on chemosynthetic ecosystems. Here we assess connectivity and population structure of two vent-associated invertebrates--the shrimp Chorocaris sp. 2 and the squat lobster Munidopsis lauensis--that are common at deep-sea hydrothermal vents in the western Pacific. While Chorocaris sp. 2 has only been observed at hydrothermal vent sites, M. lauensis can be found throughout the deep sea but occurs in higher abundance around the periphery of active vents We sequenced mitochondrial COI genes and deployed nuclear microsatellite markers for both species at three sites in Manus Basin and either North Fiji Basin (Chorocaris sp. 2) or Lau Basin (Munidopsis lauensis). We assessed genetic differentiation across a range of spatial scales, from approximately 2.5 km to more than 3000 km. Population structure for Chorocaris sp. 2 was comparable to that of the vent-associated snail Ifremeria nautilei, with a single seemingly well-mixed population within Manus Basin that is genetically differentiated from conspecifics in North Fiji Basin. Population structure for Munidopsis lauensis was more complex, with two genetically differentiated populations in Manus Basin and a third well-differentiated population in Lau Basin. The unexpectedly high level of genetic differentiation between M. lauensis populations in Manus Basin deserves further study since it has implications for conservation and management of diversity in deep-sea hydrothermal vent ecosystems.

Iterated prisoner's dilemma (IPD) researchers have shown that strong positive reputations plus an efficient reputation evaluation system encourages both sides to pursue long-term collaboration and to avoid falling into mutual defection cycles. In agent-based environments with reliable reputation rating systems, agents interested in maximizing their private interests must show concern for other agents as well as their own self-reputations--an important capability that standard IPD game agents lack. Here we present a novel learning agent model possessing self-reputation awareness. Agents in our proposed model are capable of evaluating self-behaviors based on a mix of public and private interest considerations, and of testing various solutions aimed at meeting social standards. Simulation results indicate multiple outcomes from the addition of a small percentage of self-reputation awareness agents: faster cooperation, faster movement toward stability in an agent society, a higher level of public interest in the agent society, the resolution of common conflicts between public and private interests, and a lower potential for rational individual behavior to transform into irrational group behavior.

Interferon-beta (IFN-β) treatment may not be effective in neuromyelitis optica (NMO). Whether the poor response to IFN-β is related to long spinal cord lesions (LSCL) or the NMO disease entity itself is unclear. We evaluated the spinal cord involvement of patients with multiple sclerosis (MS) and NMO, as well as the response after receiving IFN-β. Forty-nine MS and 21 NMO patients treated with IFN-β for at least 2 years from 2002-2008 were enrolled in this study and the treatment response was analyzed 2 years post-treatment. In the study, spinal cord lesions were present in 57.1% (28/49) of the MS patients, of which 16.3% (8/49) presented spinal cord lesions longer than 3 vertebral segments (LSCL). Responses to IFN-β treatment were seen in 69.3% (34/49) of all the MS cases, of which the appropriate response rates were 76.1% (16/21) in MS patients without spinal cord lesions and 37.5% (3/8) in patients with LSCL. Only 14.2% (3/21) of NMO patients responded to IFN-β treatment. In conclusion, spinal cord lesion is common in MS patients in Taiwan. Both NMO and MS patients with LSCL had a poor response to IFN-β treatment. NMO patients had a worse response to IFN-β treatment than MS patients with LSCL, which shows that the crucial structural defect is something other than LSCL such as the elevated serum IL17 level in NMO compared to MS.

The number of scholarly documents available on the web is estimated using capture/recapture methods by studying the coverage of two major academic search engines: Google Scholar and Microsoft Academic Search. Our estimates show that at least 114 million English-language scholarly documents are accessible on the web, of which Google Scholar has nearly 100 million. Of these, we estimate that at least 27 million (24%) are freely available since they do not require a subscription or payment of any kind. In addition, at a finer scale, we also estimate the number of scholarly documents on the web for fifteen fields: Agricultural Science, Arts and Humanities, Biology, Chemistry, Computer Science, Economics and Business, Engineering, Environmental Sciences, Geosciences, Material Science, Mathematics, Medicine, Physics, Social Sciences, and Multidisciplinary, as defined by Microsoft Academic Search. In addition, we show that among these fields the percentage of documents defined as freely available varies significantly, i.e., from 12 to 50%.

Prostate cancer (PCa) cells undergoing neuroendocrine differentiation (NED) are clinically relevant to the development of relapsed castration-resistant PCa. Increasing evidences show that autophagy involves in the development of neuroendocrine (NE) tumors, including PCa. To clarify the effect of autophagy on NED, androgen-sensitive PCa LNCaP cells were examined. Treatment of LNCaP cells with IL-6 resulted in an induction of autophagy. In the absence of androgen, IL-6 caused an even stronger activation of autophagy. Similar result was identified in NED induction. Inhibition of autophagy with chloroquine (CQ) markedly decreased NED. This observation was confirmed by beclin1 and Atg5 silencing experiments. Further supporting the role of autophagy in NED, we found that LC3 was up-regulated in PCa tissue that had relapsed after androgen-deprivation therapy when compared with their primary tumor counterpart. LC3 staining in relapsed PCa tissue showed punctate pattern similar to the staining of chromogranin A (CgA), a marker for NED cells. Moreover, autophagy inhibition induced the apoptosis of IL-6 induced NE differentiated PCa cells. Consistently, inhibition of autophagy by knockdown of beclin1 or Atg5 sensitized NE differentiated LNCaP cells to etoposide, a chemotherapy drug. To identify the mechanisms, phosphorylation of IL-6 downstream targets was analyzed. An increase in phospho-AMPK and a decrease in phospho-mTOR were found, which implies that IL-6 regulates autophagy through the AMPK/mTOR pathway. Most important to this study is the discovery of REST, a neuronal gene-specific transcriptional repressor that is involved in autophagy activation. REST was down-regulated in IL-6 treatment. Knockdown experiments suggest that REST is critical to NED and autophagy activation by IL-6. Together, our studies imply that autophagy is involved in PCa progression and plays a cytoprotective role when NED is induced in PCa cells by IL-6 treatment. These results reveal the potential of targeting autophagy as part of a combined therapeutic regime for NE tumors.

ARX/Arx is a homeodomain-containing transcription factor necessary for the specification and early maintenance of pancreatic endocrine α-cells. Many transcription factors important to pancreas development, including ARX/Arx, are also crucial for proper brain development. Although null mutations of ARX in human patients result in the severe neurologic syndrome XLAG (X-linked lissencephaly associated with abnormal genitalia), the most common mutation is the expansion of the first polyalanine tract of ARX, which results primarily in the clinical syndrome ISSX (infantile spasms). Mouse models of XLAG, ISSX and other human ARX mutations demonstrate a direct genotype-phenotype correlation in ARX-related neurologic disorders. Furthermore, mouse models utilizing a polyalanine tract expansion mutation have illustrated critical developmental differences between null mutations and expansion mutations in the brain, revealing context-specific defects. Although Arx is known to be required for the specification and early maintenance of pancreatic glucagon-producing α-cells, the consequences of the Arx polyalanine expansion on pancreas development remain unknown. Here we report that mice with an expansion mutation in the first polyalanine tract of Arx exhibit impaired α-cell specification and maintenance, with gradual α-cell loss due to apoptosis. This is in contrast to the re-specification of α-cells into β- and δ-cells that occurs in mice null for Arx. Overall, our analysis of an Arx polyalanine expansion mutation on pancreatic development suggests that impaired α-cell function might also occur in ISSX patients.

Genomic and proteomic analysis of normal and cancer tissues has yielded abundant molecular information for potential biomarker and therapeutic targets. Considering potential advantages in accessibility to pharmacological intervention, identification of targets resident on the vascular endothelium within tumors is particularly attractive. By employing mass spectrometry (MS) as a tool to identify proteins that are over-expressed in tumor-associated endothelium relative to normal cells, we aimed to discover targets that could be utilized in tumor angiogenesis cancer therapy. We developed proteomic methods that allowed us to focus our studies on the discovery of cell surface/secreted proteins, as they represent key antibody therapeutic and biomarker opportunities. First, we isolated endothelial cells (ECs) from human normal and kidney cancer tissues by FACS using CD146 as a marker. Additionally, dispersed human colon and lung cancer tissues and their corresponding normal tissues were cultured ex-vivo and their endothelial content were preferentially expanded, isolated and passaged. Cell surface proteins were then preferentially captured, digested with trypsin and subjected to MS-based proteomic analysis. Peptides were first quantified, and then the sequences of differentially expressed peptides were resolved by MS analysis. A total of 127 unique non-overlapped (157 total) tumor endothelial cell over-expressed proteins identified from directly isolated kidney-associated ECs and those identified from ex-vivo cultured lung and colon tissues including known EC markers such as CD146, CD31, and VWF. The expression analyses of a panel of the identified targets were confirmed by immunohistochemistry (IHC) including CD146, B7H3, Thy-1 and ATP1B3. To determine if the proteins identified mediate any functional role, we performed siRNA studies which led to previously unidentified functional dependency for B7H3 and ATP1B3.

Background: The Dietary Approaches to Stop Hypertension (DASH) diet has been recognized as effective to lower blood pressure in feeding trials, but compliance with the diet must be persistent to maximize health benefits in clinical practice. This paper reports a systematic review of the latest evidence on the method to assess DASH compliance and the corresponding patients' compliance in interventional settings., Methods: The databases including MEDLINE, EBM Reviews, EMBASE, and CINAHL Plus were searched for original research studies published in the period of January 1992-December 2012 that evaluated compliance with DASH diet. Studies written in English language, with DASH intervention, with complete documentation of the degree of DASH compliance and the assessment method used were included in this review. The search terms included: dietary approaches to stop hypertension, DASH, compliance, adherence, consistency, and concordance., Results: Nine studies were included. Different types of interventions were identified, ranging from feeding trial to dietary counseling. These studies differed in the assessment methods used to evaluate DASH compliance, which included objective approaches like measurement of urinary excretion, and subjective approaches like dietary intake assessment for DASH target comparison and construction of DASH scoring systems. Compliance levels were lower in educational interventions than that of the original DASH feeding trial., Conclusions: To conclude, although no consensus existed regarding the best approach to assess DASH compliance, its suboptimal compliance warrants attention. This study implied a need to investigate effective approaches to sustain the DASH dietary pattern beyond counselling alone.

Soluble human leukocyte antigen-G (HLA-G) is a non-classical class Ib HLA molecule that is secreted from blastocysts. Soluble HLA-G modulates the immune tolerance of the mother and can be used as a prognostic factor for the clinical pregnancy rate. However, the underlying mechanism of how soluble HLA-G5 affects pregnancy remains largely unknown. We hypothesized that soluble HLA-G5 promotes successful implantation and pregnancy by modulating trophoblast invasion through receptor binding and activation of extracellular signal-regulated protein kinase (ERK) signaling pathway. Recombinant HLA-G5 protein over-expressed in E. coli BL21 was purified to near homogeneity. We studied the expression of HLA-G5 and its receptors, the leukocyte immunoglobulin-like receptor subfamily B1 (LILRB1) and killer cell immunoglobulin-like receptor 2DL4 (KIR2DL4), in primary trophoblasts and trophoblastic (JAr and JEG-3) cell lines by florescence-labeled HLA-G5. HLA-G5 was detected in the primary trophoblasts and JEG-3 cells. The LILRB1 and KIR2DL4 receptors were expressed in both primary trophoblasts and trophoblastic cell lines. HLA-G5 stimulated cell invasion (p<0.05) and increased urokinase (uPA) and matrix metalloproteinases (MMPs) transcripts and their activity (p<0.05) in trophoblastic cells. HLA-G5 activated the ERK signaling pathway and induced ERK1/2 phosphorylation in the trophoblastic cell lines. Addition of ERK inhibitors (U0126 and PD98059) nullified the stimulatory effect of HLA-G5 on trophoblastic cell invasion. Taken together, HLA-G5 induced trophoblast invasion by binding to KIR2DL4 and LILRB1, by increasing uPA and MMPs expressions and by activating the ERK signaling pathway.

Background: Alkylphenols, such as nonylphenol (NP) and 4-octylphenol (4-OP), have the potential to disturb immune system due to their weak estrogen-like activity, an effect with potential serious public health impact due to the worldwide distribution of these substances. Plasmacytoid dendritic cells (PDCs) can secrete large amounts of type I IFNs and are critical in immune regulation. However, there has been limited study about the influence of alkylphenols on the function of pDCs., Objective: The aim of this study was to examine the effect of alkylphenols on pDC functions in vitro and in vivo and then further explored the involved signaling pathways and epigenetic changes., Methods: Circulating pDCs from human peripheral blood mononuclear cells were treated with alkylphenols with or without CpG stimulation. Alkylphenol-associated cytokine responses, signaling events, histone modifications and viral activity were further examined. In NP-exposed mice, the effect of NP on splenic pDC function and allergic lung inflammation were also assessed., Results: The results showed that NP increased the expression of TNF-α, but suppressed IL-10 production in the range of physiological doses, concomitant with activation of the MKK3/6-p38 signaling pathway and enhanced levels of acetylated histone 3 as well as histone 4 at the TNFA gene locus. Further, in CpG-stimulated pDCs, NP suppressed type I IFNs production, associated with down-regulation of IRF-7 and MKK1/2-ERK-Elk-1 pathways and led to the impaired anti-enterovirus 71 activity in vitro. Additionally, splenic pDCs from NP-exposed mice showed similar cytokine changes upon CpG stimulation under conditions relevant to route and level of exposure in humans. NP treatment also enhanced allergic lung inflammation in vivo., Conclusion: Alkylphenols may influence pDCs' functions via their abilities to induce expression of a pro-inflammatory cytokine, TNF-α, and to suppress regulatory cytokines, including IL-10, IFN-α and IFN-β, suggesting the potential impact of endocrine disrupting chemicals on immune regulation.

Background: The development and progression of colorectal cancer (CRC) involve a complex process of multiple genetic changes. Tumor suppressor p53 is capable of determining the fate of CRC cells. However, the role of a p53-inducible modulator, ribosomal protein S27-like (RPS27L), in CRC is unknown., Methods: Here, the differential expression of RPS27L was examined in the feces and colonic tissues of CRC patients, to explore its possible correlation with patient survival and to investigate the cellular mechanisms underlying their clinical outcomes. Eighty intermediate-stage CRC patients (42 at stage II and 38 at stage III) were divided into two groups according to their fecal RPS27L mRNA levels. The survival probabilities of the groups were estimated using the Kaplan-Meier method. The RPS27L protein in the colonic tissues of stage III patients with different prognoses was further examined immunohistochemically. RPS27L expression in LoVo cells was manipulated to examine the possible cellular responses in vitro., Results: Elevated RPS27L expression, in either feces or tissues, was related to a better prognosis. In vitro, RPS27L-expressing LoVo cells ceased DNA synthesis and apoptotic activity while the expression of their DNA repair molecules was upregulated., Conclusions: Elevated RPS27L may improve the prognoses of certain CRC patients by enhancing the DNA repair capacity of their colonic cells, and can be determined in feces. By integrating clinical, molecular, and cellular data, our study demonstrates that fecal RPS27L may be a useful index for predicting prognoses and guiding personalized therapeutic strategies, especially in patients with intermediate-stage CRC.

The specification and differentiation of pancreatic endocrine cell populations (α-, β-, δ, PP- and ε-cells) is orchestrated by a combination of transcriptional regulators. In the pancreas, Aristaless-related homeobox gene (Arx) is expressed first in the endocrine progenitors and then restricted to glucagon-producing α-cells. While the functional requirement of Arx in early α-cell specification has been investigated, its role in maintaining α-cell identity has yet to be explored. To study this later role of Arx, we have generated mice in which the Arx gene has been ablated specifically in glucagon-producing α-cells. Lineage-tracing studies and immunostaining analysis for endocrine hormones demonstrate that ablation of Arx in neonatal α-cells results in an α-to-β-like conversion through an intermediate bihormonal state. Furthermore, these Arx-deficient converted cells express β-cell markers including Pdx1, MafA, and Glut2. Surprisingly, short-term ablation of Arx in adult mice does not result in a similar α-to-β-like conversion. Taken together, these findings reveal a potential temporal requirement for Arx in maintaining α-cell identity.

We performed a genome wide analysis of 164 urothelial carcinoma samples and 27 bladder cancer cell lines to identify copy number changes associated with disease characteristics, and examined the association of amplification events with stage and grade of disease. Multiplex inversion probe (MIP) analysis, a recently developed genomic technique, was used to study 80 urothelial carcinomas to identify mutations and copy number changes. Selected amplification events were then analyzed in a validation cohort of 84 bladder cancers by multiplex ligation-dependent probe assay (MLPA). In the MIP analysis, 44 regions of significant copy number change were identified using GISTIC. Nine gene-containing regions of amplification were selected for validation in the second cohort by MLPA. Amplification events at these 9 genomic regions were found to correlate strongly with stage, being seen in only 2 of 23 (9%) Ta grade 1 or 1-2 cancers, in contrast to 31 of 61 (51%) Ta grade 3 and T2 grade 2 cancers, p<0.001. These observations suggest that analysis of genomic amplification of these 9 regions might help distinguish non-invasive from invasive urothelial carcinoma, although further study is required. Both MIP and MLPA methods perform well on formalin-fixed paraffin-embedded DNA, enhancing their potential clinical use. Furthermore several of the amplified genes identified here (ERBB2, MDM2, CCND1) are potential therapeutic targets.

In a structure-activity relationship (SAR) study, 3-methoxy-1,4-phenanthrenequinones, calanquinone A (6a), denbinobin (6b), 5-OAc-calanquinone A (7a) and 5-OAc-denbinobin (7b), have significantly promising cytotoxicity against various human cancer cell lines (IC(50) 0.08-1.66 µg/mL). Moreover, we also established a superior pharmacophore model for cytotoxicity (r = 0.931) containing three hydrogen bond acceptors (HBA1, HBA2 and HBA3) and one hydrophobic feature (HYD) against MCF-7 breast cancer cell line. The pharmacophore model indicates that HBA3 is an essential feature for the oxygen atom of 5-OH in 6a-b and for the carbonyl group of 5-OCOCH(3) in 7a-b, important for their cytotoxic properties. The SAR for moderately active 5a-b (5-OCH(3)), and highly active 6a-b and 7a-b, are also elaborated in a spatial aspect model. Further rational design and synthesis of new cytotoxic phenanthrene analogs can be implemented via this model. Additionally, employing a ChemGPS-NP based model for cytotoxicity mode of action (MOA) provides support for a preliminary classification of compounds 6a-b as topoisomerase II inhibitors.

Renal angiomyolipoma are part of the PEComa family of neoplasms, and occur both in association with Tuberous Sclerosis Complex (TSC) and independent of that disorder. Previous studies on the molecular genetic alterations that occur in angiomyolipoma are very limited. We evaluated 9 angiomyolipoma for which frozen tissue was available from a consecutive surgical series. Seven of 8 samples subjected to RT-PCR-cDNA sequencing showed mutations in TSC2; none showed mutations in TSC1 or RHEB. Six of the seven mutations were deletions. We searched for 983 activating and inactivating mutations in 115 genes, and found none in these tumors. Similarly analysis for genomic regions of loss or gain, assessed by Affymetrix SNP6.0 analysis, showed no abnormalities. Loss of heterozygosity in the TSC2 region was commonly seen, except in patients with low frequency TSC2 mutations. We conclude that sporadic renal angiomyolipoma usually have mutations in TSC2, but not TSC1 or RHEB, and have no other common genomic events, among those we searched for. However, chromosomal translocations and gene fusion events were not assessed here. TSC2 inactivation by mutation is a consistent and likely necessary genetic event in the pathogenesis of most angiomyolipoma.

Protoapigenone, a natural flavonoid possessing an unusual p-quinol moiety on its B-ring, is a novel prospective anticancer agent with low toxicity that is currently in development. The first economical, one-step synthesis of protoapigenone from apigenin is described on up to gram scale. 13 new 1'-O-alkylflavone analogs were also synthesized, either from apigenin or β-naphthoflavone. The in vitro cytotoxic activity of each compound was tested on six human cancer cell lines (HepG2, Hep3B, Ca9-22, A549, MCF-7 and MDA-MB-231). In the case of 1'-O-alkyl-protoapigenone derivatives, structure-activity relationships were found depending on the side-chain, and protoapigenone 1'-O-butyl ether was found to exert significantly stronger activity against three of the cell lines (Hep3B, MCF-7 and MDA-MB-231) than its non-substituted analog, protoapigenone itself. In contrast to this, all β-naphthoflavone derivatives bearing the same pharmacophore on their B-ring showed decreased cytotoxic activities when substituted with an O-alkyl side-chain at position 1', comparing to that of the non-substituted compound.

Background: The signaling pathways imposing hormonal control over adipocyte differentiation are poorly understood. While insulin and Akt signaling have been found previously to be essential for adipogenesis, the relative importance of their many downstream branches have not been defined. One direct substrate that is inhibited by Akt-mediated phosphorylation is the tuberous sclerosis complex 2 (TSC2) protein, which associates with TSC1 and acts as a critical negative regulator of the mammalian target of rapamycin (mTOR) complex 1 (mTORC1). Loss of function of the TSC1-TSC2 complex results in constitutive mTORC1 signaling and, through mTORC1-dependent feedback mechanisms and loss of mTORC2 activity, leads to a concomitant block of Akt signaling to its other downstream targets., Methodology/principal Findings: We find that, despite severe insulin resistance and the absence of Akt signaling, TSC2-deficient mouse embryo fibroblasts and 3T3-L1 pre-adipocytes display enhanced adipocyte differentiation that is dependent on the elevated mTORC1 activity in these cells. Activation of mTORC1 causes a robust increase in the mRNA and protein expression of peroxisome proliferator-activated receptor gamma (PPARgamma), which is the master transcriptional regulator of adipocyte differentiation. In examining the requirements for different Akt-mediated phosphorylation sites on TSC2, we find that only TSC2 mutants lacking all five previously identified Akt sites fully block insulin-stimulated mTORC1 signaling in reconstituted Tsc2 null cells, and this mutant also inhibits adipogenesis. Finally, renal angiomyolipomas from patients with tuberous sclerosis complex contain both adipose and smooth muscle-like components with activated mTORC1 signaling and elevated PPARgamma expression., Conclusions/significance: This study demonstrates that activation of mTORC1 signaling is a critical step in adipocyte differentiation and identifies TSC2 as a primary target of Akt driving this process. Therefore, the TSC1-TSC2 complex regulates the differentiation of mesenchymal cell lineages, at least in part, through its control of mTORC1 activity and PPARgamma expression.