The structure of importin α and the nuclear localization peptide of ChREBP, and small compound inhibitors of ChREBP-importin α interactions.
The carbohydrate response element binding protein (ChREBP) is a glucose-responsive transcription factor that plays a critical role in glucose-mediated induction of genes involved in hepatic glycolysis and lipogenesis. In response to fluctuating blood glucose levels ChREBP activity is regulated mainly by nucleocytoplasmic shuttling of ChREBP. Under high glucose ChREBP binds to importin α and importin β and translocates into the nucleus to initiate transcription. We have previously shown that the nuclear localization signal site (NLS) for ChREBP is bipartite with the NLS extending from Arg158 to Lys190. Here, we report the 2.5 Å crystal structure of the ChREBP-NLS peptide bound to importin α. The structure revealed that the NLS binding is monopartite, with the amino acid residues K171RRI174 from the ChREBP-NLS interacting with ARM2-ARM5 on importin α. We discovered that importin α also binds to the primary binding site of the 14-3-3 proteins with high affinity, which suggests that both importin α and 14-3-3 are each competing with the other for this broad-binding region (residues 117-196) on ChREBP. We screened a small compound library and identified two novel compounds that inhibit the ChREBP-NLS/importin α interaction, nuclear localization, and transcription activities of ChREBP. These candidate molecules support developing inhibitors of ChREBP that may be useful in treatment of obesity and the associated diseases.


Clinical Activity of the γ-Secretase Inhibitor PF-03084014 in Adults With Desmoid Tumors (Aggressive Fibromatosis).
Purpose Desmoid tumors (aggressive fibromatosis) arise from connective tissue cells or fibroblasts. In general, they are slow growing and do not metastasize; however, locally aggressive desmoid tumors can cause severe morbidity and loss of function. Disease recurrence after surgery and/or radiation and diagnosis of multifocal desmoid tumors highlight the need to develop effective systemic treatments for this disease. In this study, we evaluate objective response rate after therapy with the γ-secretase inhibitor PF-03084014 in patients with recurrent, refractory, progressive desmoid tumors. Patients and Methods Seventeen patients with desmoid tumors received PF-03084014 150 mg orally twice a day in 3-week cycles. Response to treatment was evaluated at cycle 1 and every six cycles, that is, 18 weeks, by RECIST (Response Evaluation Criteria in Solid Tumors) version 1.1. Patient-reported outcomes were measured at baseline and at every restaging visit by using the MD Anderson Symptoms Inventory. Archival tumor and blood samples were genotyped for somatic and germline mutations in APC and CTNNB1. Results Of 17 patients accrued to the study, 15 had mutations in APC or CTNNB1 genes. Sixteen patients (94%) were evaluable for response; five (29%) experienced a confirmed partial response and have been on study for more than 2 years. Another five patients with prolonged stable disease as their best response remain on study. Patient-reported outcomes confirmed clinician reporting that the investigational agent was well tolerated and, in subgroup analyses, participants who demonstrated partial response also experienced clinically meaningful and statistically significant improvements in symptom burden. Conclusion PF-03084014 was well tolerated and demonstrated promising clinical benefit in patients with refractory, progressive desmoid tumors who receive long-term treatment.


Single subunit chimeric integrins as mimics and inhibitors of endogenous integrin functions in receptor localization, cell spreading and migration, and matrix assembly.
The ability of single subunit chimeric receptors containing various integrin beta intracellular domains to mimic and/or inhibit endogenous integrin function was examined. Chimeric receptors consisting of the extracellular and transmembrane domains of the small subunit of the human interleukin-2 receptor connected to either the beta 1, beta 3, beta 3B, or beta 5 intracellular domain were transiently expressed in normal human fibroblasts. When expressed at relatively low levels, the beta 3 and beta 5 chimeras mimicked endogenous ligand-occupied integrins and, like the beta 1 chimera (LaFlamme, S. E., S. K. Akiyama, and K. M. Yamada. 1992. J. Cell Biol. 117:437), concentrated with endogenous integrins in focal adhesions and sites of fibronectin fibril formation. In contrast, the chimeric receptor containing the beta 3B intracellular domain (a beta 3 intracellular domain modified by alternative splicing) was expressed diffusely on the cell surface, indicating that alternative splicing can regulate integrin receptor distribution by an intracellular mechanism. Furthermore, when expressed at higher levels, the beta 1 and beta 3 chimeric receptors functioned as dominant negative mutants and inhibited endogenous integrin function in localization to fibronectin fibrils, fibronectin matrix assembly, cell spreading, and cell migration. The beta 5 chimera was a less effective inhibitor, and the beta 3B chimera and the reporter lacking an intracellular domain did not inhibit endogenous integrin function. Comparison of the relative levels of expression of the transfected beta 1 chimera and the endogenous beta 1 subunit indicated that in 10 to 15 h assays, the beta 1 chimera can inhibit cell spreading when expressed at levels approximately equal to the endogenous beta 1 subunit. Levels of chimeric receptor expression that inhibited cell spreading also inhibited cell migration, whereas lower levels were able to inhibit alpha 5 beta 1 localization to fibrils and matrix assembly. Our results indicate that single subunit chimeric integrins can mimic and/or inhibit endogenous integrin receptor function, presumably by interacting with cytoplasmic components critical for endogenous integrin function. Our results also demonstrate that beta intracellular domains, expressed in this context, display specificity in their abilities to mimic and inhibit endogenous integrin function. Furthermore, the approach that we have used permits the analysis of intracellular domain function in the processes of cell spreading, migration and extracellular matrix assembly independent of effects due to the rest of integrin dimers. This approach should prove valuable in the further analysis of integrin intracellular domain function in these and other integrin-mediated processes requiring the interaction of integrins with cytoplasmic components.


Noncoding RNA in development.
Non-protein-coding sequences increasingly dominate the genomes of multicellular organisms as their complexity increases, in contrast to protein-coding genes, which remain relatively static. Most of the mammalian genome and indeed that of all eukaryotes is expressed in a cell- and tissue-specific manner, and there is mounting evidence that much of this transcription is involved in the regulation of differentiation and development. Different classes of small and large noncoding RNAs (ncRNAs) have been shown to regulate almost every level of gene expression, including the activation and repression of homeotic genes and the targeting of chromatin-remodeling complexes. ncRNAs are involved in developmental processes in both simple and complex eukaryotes, and we illustrate this in the latter by focusing on the animal germline, brain, and eye. While most have yet to be systematically studied, the emerging evidence suggests that there is a vast hidden layer of regulatory ncRNAs that constitutes the majority of the genomic programming of multicellular organisms and plays a major role in controlling the epigenetic trajectories that underlie their ontogeny.


Maternal vitamin D status in pregnancy is associated with adiposity in the offspring: findings from the Southampton Women's Survey.
Low vitamin D status has been linked to adiposity, but little is known of the effects of low status in pregnancy on offspring body composition. The objective was to determine how maternal vitamin D status relates to lean and fat mass of the offspring. The offspring of 977 pregnant women, who had serum 25-hydroxyvitamin D [25(OH)D] measured at 34 wk gestation, were followed up within 3 wk of birth and at 4 and 6 y of age for dual-energy X-ray absorptiometry assessment of lean and fat mass. The median maternal serum 25(OH)D concentration was 62 nmol/L (IQR: 43-89 nmol/L); 35% of the women studied had values <50 nmol/L. Lower vitamin D status was associated with lower fat mass in the offspring at birth but with greater fat mass at ages 4 and 6 y. It was not associated with lean mass at any of the ages studied. The opposing associations seen between maternal 25(OH)D (SDs) and fat mass (SDs) in the offspring at birth and at age 6 y were robust to adjustment for a range of confounding factors, including maternal BMI and weight gain in pregnancy [β (95% CI): 0.08 (0.02, 0.15) and -0.10 (-0.17, -0.02), respectively]. The key independent predictors of higher maternal vitamin D status were season of assessment and use of vitamin D supplements. Lower maternal vitamin D status may be linked to programmed differences in offspring fat mass. The findings require replication but add to a growing evidence base for a role of vitamin D in the origins of adiposity.


Group schema therapy for eating disorders: study protocol.
The treatment of eating disorders is a difficult endeavor, with only a relatively small proportion of clients responding to and completing standard cognitive behavioural therapy (CBT). Given the prevalence of co-morbidity and complex personality traits in this population, Schema Therapy has been identified as a potentially viable treatment option. A case series of Group Schema Therapy for Eating Disorders (ST-E-g) yielded positive findings and the study protocol outlined in this article aims to extend upon these preliminary findings to evaluate group Schema Therapy for eating disorders in a larger sample (n = 40). Participants undergo a two-hour assessment where they complete a number of standard questionnaires and their diagnostic status is ascertained using the Eating Disorder Examination. Participants then commence treatment, which consists of 25 weekly group sessions lasting for 1.5 h and four individual sessions. Each group consists of five to eight participants and is facilitated by two therapists, at least one of who is a registered psychologist trained on schema therapy. The primary outcome in this study is eating disorder symptom severity. Secondary outcomes include: cognitive schemas, self-objectification, general quality of life, self-compassion, schema mode presentations, and Personality Disorder features. Participants complete psychological measures and questionnaires at pre, post, six-month and 1-year follow-up. This study will expand upon preliminary research into the efficacy of group Schema Therapy for individuals with eating disorders. If group Schema Therapy is shown to reduce eating disorder symptoms, it will hold considerable promise as an intervention option for a group of disorders that is typically difficult to treat. ACTRN12615001323516. Registered: 2/12/2015 (retrospectively registered, still recruiting).


Cardiorespiratory fitness, alcohol, and mortality in men: the Cooper Center longitudinal study.
Studies have found that higher levels of cardiorespiratory fitness and light to moderate alcohol intake reduce the risk for premature death. Scant evidence, however, exists assessing the joint effects of both measures on all-cause and cardiovascular disease (CVD) mortality. This study aims to examine the independent and joint effects of alcohol consumption and cardiorespiratory fitness on all-cause and cardiovascular-related mortality in a large cohort of men. This prospective study included 29,402 men who came to the Cooper Clinic (Dallas, TX) for a preventive medicine visit from 1973 to 2006. Data were analyzed in 2011. The primary exposure variables were tertiles of cardiorespiratory fitness and four categories of alcohol consumption, and the outcomes were all-cause and CVD mortality. Cox proportional hazards regression was used to model the association between alcohol intake, cardiorespiratory fitness, and all-cause and CVD mortality, controlling for potential confounders. A total of 1830 (all-cause) and 523 (CVD) deaths occurred in men over an average follow-up period of 17.4 years (SD=9.1). A linear relationship was observed (p<0.001) between increased fitness and reduced all-cause and CVD mortality. Specifically, moderate and high levels of fitness reduced the risk for all-cause mortality (HR=0.67, 95% CI=0.60, 0.74, and HR=0.57, 95% CI=0.49, 0.67, respectively) and CVD mortality in comparison to the low-fitness reference group (HR=0.70, 95% CI=0.57, 0.85; HR=0.54, 95% CI=0.40, 0.75, respectively), while controlling for alcohol intake and other covariates. A significant curvilinear relationship was found (p=0.01) between alcohol intake and all-cause mortality (but not CVD mortality), while controlling for fitness and other covariates. In a categoric examination of alcohol intake and mortality, adjusting for fitness and other confounders, there was no statistically significant effect of light drinking compared to heavy drinking on all-cause mortality or CVD mortality. An examination of the joint effects of fitness and alcohol on all-cause mortality showed that moderate and high fitness levels were protective against mortality irrespective of alcohol consumption levels. Few significant combined effects for CVD mortality reduction were found. Alcohol consumption did not significantly modify the association between fitness and mortality in this large cohort of men.


The social and economic determinants of smoking in Moscow, Russia.
Despite a high prevalence of smoking for decades, recent research has documented an increase in the rates of both male and female smoking in post-Soviet Russia. As yet, however, little research has taken place on smoking at the subnational level. The current study addresses this deficit by examining smoking in Moscow -- the city that has been at the forefront of the entry into the Russian market of transnational tobacco corporations (TTCs) in the transition period. Data were obtained from the Moscow Health Survey 2004 -- a stratified random sample of 1190 people representative of Moscow's larger population. Information was obtained about subjects' smoking habits and age of smoking initiation. The prevalence of smoking was high among both men (55.5%) and women (26.9%), with significantly higher rates in the younger age groups. There was also a high prevalence of smoking initiation before age 15 years, especially in the youngest women (18-30 years). Logistic regression analysis showed that respondents' age, binge drinking, locus of control and economic situation were important determinants of smoking. Although lifestyle factors seem to underpin the generally high levels of smoking, other things, such as its high prevalence in the younger generations and the factors associated with smoking (locus of control), nevertheless suggest that the TTCs may have played an important role in the spread of smoking in transitional Russia's changing social environment.


Stomal recurrence. A critical analysis of risk factors.
Stomal recurrence developed in 5% of the 507 patients who underwent total laryngectomy for a squamous cell carcinoma. The most common site of the primary tumor was the glottis, followed by the supraglottic and pyriform sinus regions. Initial subglottic extension of the tumor and metastatic lymphadenopathy were the most significant risk factors. The primary tumor size, prior emergency tracheostomy, and conservation surgical procedures had no effect on the incidence of stomal recurrence. The median survival in patients with stomal malignant neoplasms was only five months. In high-risk patients, extended dissection or elective postoperative radiotherapy is recommended.


Population stratification confounds genetic association studies among Latinos.
In the United States, asthma prevalence and mortality are the highest among Puerto Ricans and the lowest among Mexicans. Case-control association studies are a powerful strategy for identifying genes of modest effect in complex diseases. However, studies of complex disorders in admixed populations such as Latinos may be confounded by population stratification. We used ancestry informative markers (AIMs) to identify and correct for population stratification among Mexican and Puerto Rican subjects participating in case-control studies of asthma. Three hundred and sixty-two subjects with asthma (Mexican: 181, Puerto Rican: 181) and 359 ethnically matched controls (Mexican: 181, Puerto Rican: 178) were genotyped for 44 AIMs. We observed a greater than expected degree of association between pairs of AIMs on different chromosomes in Mexicans (P < 0.00001) and Puerto Ricans (P < 0.00002) providing evidence for population substructure and/or recent admixture. To assess the effect of population stratification on association studies of asthma, we measured differences in genetic background of cases and controls by comparing allele frequencies of the 44 AIMs. Among Puerto Ricans but not in Mexicans, we observed a significant overall difference in allele frequencies between cases and controls (P = 0.0002); of 44 AIMs tested, 8 (18%) were significantly associated with asthma. However, after adjustment for individual ancestry, only two of these markers remained significantly associated with the disease. Our findings suggest that empirical assessment of the effects of stratification is critical to appropriately interpret the results of case-control studies in admixed populations.


Chronic rhinitis and its association with headache frequency and disability in persons with migraine: results of the American Migraine Prevalence and Prevention (AMPP) Study.
Rhinitis is a comorbidity of migraine, but its relationship to migraine headache frequency and headache-related disability is unknown. To determine if rhinitis and its subtypes are associated with an increased frequency and associated disability of migraine. The AMPP Study is a longitudinal study of individuals with "severe" headache from the US population. Respondents meeting ICHD-2 criteria for migraine in 2008 were identified and the presence of rhinitis was determined using the European Community Respiratory Health Survey (ECRHS). Those with rhinitis were subtyped as allergic, non-allergic, mixed and unclassified based on a rhinitis questionnaire. The primary outcome measures were categories of headache-day frequency and headache-related disability as measured by the Migraine Disability Assessment Scale (MIDAS). Logistic regression for ordered categories was used for modeling each outcome separately, adjusted for sociodemographics profile, headache features, headache treatments and comorbidities. The AMPP Study questionnaire was mailed to 17,892 persons and returned by 60.1% of respondents. Among the migraine sample ( N = 5849), 66.8% had rhinitis with mixed rhinitis as the most common form. The presence of rhinitis of any type was associated with headache frequency after adjusting for sociodemographic variables only (OR 1.33; 95% CI 1.16, 1.53) and in the fully adjusted model (OR 1.25; 95% CI 1.05-1.49). Headache-related disability (MIDAS category) was associated with rhinitis after adjusting for sociodemographic features (OR 1.30; 95% CI 1.17-1.46), but lost significance in the fully adjusted model (OR 1.10; 95% CI 0.96-1.26). Mixed rhinitis was associated with an increased headache frequency category in the model adjusted for sociodemographics (OR 1.45; 95% CI 1.24-1.70) and in that adjusted for all covariates (OR 1.28; 95% CI 1.05-1.57). The odds ratio for MIDAS categories were similarly increased in both models for the mixed rhinitis group. The frequency and disability of migraine are higher in persons with rhinitis, particularly those with mixed rhinitis. These results, however, should be considered preliminary until confirmed in future studies because of the modest questionnaire response rate in this study.


Comparative Analysis of Peripheral Corneal Geometry in Health and Keratoconus.
To describe and compare corneal peripheral angles in normal and keratoconic eyes, to gain a better understanding of the topography of the periphery of the cornea in keratoconus and assist practitioners in the selection and fitting of large diameter contact lenses. Eighty-eight eyes were included in the study, divided into three groups: healthy (A0, 28 eyes), keratoconus at stage I according to the Amsler-Krumeich classification (AI, 33 eyes) and keratoconus at stages II to IV (AII, 27 eyes). The Pentacam Scheimpflug system was used to manually measure the corneal peripheral angles corresponding to a chord length range between 8.6 and 12.6 mm at 8 different peripheral locations. The peripheral angle was influenced by ocular condition and by the peripheral location, with no interaction effect between both factors. Statistically significant differences were found in mean corneal peripheral angles between groups A0 (30.84°±2.33°) and AI (31.63°±2.02°) (P=0.001) and between A0 and AII (31.37°±2.11°) (P=0.030). The differences between AI and AII were not significant. In all eyes, the largest and smallest peripheral angles were found at the temporal inferior and temporal superior locations, respectively, with a mean difference between largest and smallest of 3.37°±1.42° in healthy eyes and 2.96°±1.54° in keratoconus (AI+AII). Clinically insignificant differences were found in peripheral angles between keratoconus and healthy eyes, giving support to the use of large diameter, intralimbal contact lenses with peripheral designs, and resting on the same corneal region, as those fitted on normal corneas.


SCREENED: A Multistage Model of Thyroid Gland Function for Screening Endocrine-Disrupting Chemicals in a Biologically Sex-Specific Manner.
Endocrine disruptors (EDs) are chemicals that contribute to health problems by interfering with the physiological production and target effects of hormones, with proven impacts on a number of endocrine systems including the thyroid gland. Exposure to EDs has also been associated with impairment of the reproductive system and incidence in occurrence of obesity, type 2 diabetes, and cardiovascular diseases during ageing. SCREENED aims at developing in vitro assays based on rodent and human thyroid cells organized in three different three-dimensional (3D) constructs. Due to different levels of anatomical complexity, each of these constructs has the potential to increasingly mimic the structure and function of the native thyroid gland, ultimately achieving relevant features of its 3D organization including: 1) a 3D organoid based on stem cell-derived thyrocytes, 2) a 3D organoid based on a decellularized thyroid lobe stromal matrix repopulated with stem cell-derived thyrocytes, and 3) a bioprinted organoid based on stem cell-derived thyrocytes able to mimic the spatial and geometrical features of a native thyroid gland. These 3D constructs will be hosted in a modular microbioreactor equipped with innovative sensing technology and enabling precise control of cell culture conditions. New superparamagnetic biocompatible and biomimetic particles will be used to produce "magnetic cells" to support precise spatiotemporal homing of the cells in the 3D decellularized and bioprinted constructs. Finally, these 3D constructs will be used to screen the effect of EDs on the thyroid function in a unique biological sex-specific manner. Their performance will be assessed individually, in comparison with each other, and against in vivo studies. The resulting 3D assays are expected to yield responses to low doses of different EDs, with sensitivity and specificity higher than that of classical 2D in vitro assays and animal models. Supporting the "Adverse Outcome Pathway" concept, proteogenomic analysis and biological computational modelling of the underlying mode of action of the tested EDs will be pursued to gain a mechanistic understanding of the chain of events from exposure to adverse toxic effects on thyroid function. For future uptake, SCREENED will engage discussion with relevant stakeholder groups, including regulatory bodies and industry, to ensure that the assays will fit with purposes of ED safety assessment. In this project review, we will briefly discuss the current state of the art in cellular assays of EDs and how our project aims at further advancing the field of cellular assays for EDs interfering with the thyroid gland.


CD31 (PECAM-1) exists as a dimer and is heavily N-glycosylated.
CD31 (PECAM-1) is a highly abundant cell surface glycoprotein expressed on hemopoietic and endothelial cells where it functions as a homophilic adhesion and signaling receptor. Since dimerization and appropriate glycosylation are important features in the regulation of cell surface interactions and signal transduction, we studied the pattern of glycosylation as well as the ability of CD31 to undergo dimerization, both in solution and when expressed on cell membranes. CD31 is heavily glycosylated, with an approximate carbohydrate content of 21%. Nineteen neutral and thirteen sialylated glycans were identified. Ultracentrifugation analysis showed that soluble recombinant CD31 exists in equilibrium between a monomer and a dimer with an approximate dissociation constant of 12.5 microM. Chemical cross-linking studies of both soluble and membrane-expressed CD31 confirmed that CD31 exists as a dimer. These studies suggest that, like E-cadherin, PECAM-dimerization is likely to play a role in CD31 adhesion and signaling.


A review on citation amnesia in depression and inflammation research.
Once original scientific results are published the author has the "intellectual property" and may claim ownership. Discovery credit is one of the most important "rewards" for scientists and thus incorrect credits undermine the reward system of science. Scientists who publish should therefore give proper credit and acknowledge the primary sources. Failure to do so is regarded as "citation negligence", "the disregard syndrome", "citation amnesia", "plagiarism by omission", "bibliographic plagiarism" or "citation plagiarism", and may range from an unconscious or conscious "failure to credit a prior discoverer so as to give an improper impression of priority" to "the appropriation of another person's ideas or results without given proper credit". False discovery credit is considered to be "a menace to honest science", "a serious transgression" or "intellectual theft, be it intentional or not". This paper describes some examples of citation amnesia showing that scientists often fail to credit prior sources and give false discovery credit to other scientists. One example is the association between major depression and activated immuno-inflammatory pathways, a discovery by European groups and published in many papers since 1990. Now, 25 years later, it is commonplace that these theories are credited to secondary American sources whose work in "the last decade", did or did not examine these pathways in major depression. This gives an improper impression of priority of American-based scientists. Here it is proposed that this citation amnesia and plagiarism reinforced the wrong science and had negative effects on the development of immune-inflammatory biomarkers and new immune-related treatments for depression. It is concluded that journal editors should improve their citation standards to guarantee correct assignment of discovery credit for example by demanding a signed pledge from the authors that correct citations to the primary sources were made.


The regulation of cognitive enhancement devices: extending the medical model.
This article presents a model for regulating cognitive enhancement devices (CEDs). Recently, it has become very easy for individuals to purchase devices which directly modulate brain function. For example, transcranial direct current stimulators are increasingly being produced and marketed online as devices for cognitive enhancement. Despite posing risks in a similar way to medical devices, devices that do not make any therapeutic claims do not have to meet anything more than basic product safety standards. We present the case for extending existing medical device legislation to cover CEDs. Medical devices and CEDs operate by the same or similar mechanisms and pose the same or similar risks. This fact coupled with the arbitrariness of the line between treatment and enhancement count in favour of regulating these devices in the same way. In arguing for this regulatory model, the paper highlights potential challenges to its implementation, and suggests solutions.


Polyadenylation and beyond: emerging roles for noncanonical poly(A) polymerases.
The addition of nontemplated nucleotides, particularly adenylyl and uridylyl residues, to the 3' ends of RNA substrates has been the focus of much attention in recent years, and these studies have generated some intriguing surprises. In addition to the well-known canonical poly(A) polymerase (PAP) that polyadenylates mRNAs prior to export from the nucleus to the cytoplasm, a separate class of noncanonical poly(A) polymerases has emerged over the past decade. Studies on various organisms have led to the realization that these noncanonical PAPs, which are conserved from yeast to mammals, play crucial and diverse roles in the regulation of gene expression. Here we review the current knowledge of these enzymes, with an emphasis on the human proteins, and highlight recent discoveries that have implications far beyond the understanding of RNA metabolism itself.


Understanding the Warburg effect: the metabolic requirements of cell proliferation.
In contrast to normal differentiated cells, which rely primarily on mitochondrial oxidative phosphorylation to generate the energy needed for cellular processes, most cancer cells instead rely on aerobic glycolysis, a phenomenon termed "the Warburg effect." Aerobic glycolysis is an inefficient way to generate adenosine 5'-triphosphate (ATP), however, and the advantage it confers to cancer cells has been unclear. Here we propose that the metabolism of cancer cells, and indeed all proliferating cells, is adapted to facilitate the uptake and incorporation of nutrients into the biomass (e.g., nucleotides, amino acids, and lipids) needed to produce a new cell. Supporting this idea are recent studies showing that (i) several signaling pathways implicated in cell proliferation also regulate metabolic pathways that incorporate nutrients into biomass; and that (ii) certain cancer-associated mutations enable cancer cells to acquire and metabolize nutrients in a manner conducive to proliferation rather than efficient ATP production. A better understanding of the mechanistic links between cellular metabolism and growth control may ultimately lead to better treatments for human cancer.


You cannot tell a book by looking at the cover: Cryptic complexity in bacterial evolution.
Do genetically closely related organisms under identical, but strong selection pressure converge to a common resistant genotype or will they diverge to different genomic solutions? This question gets at the heart of how rough is the fitness landscape in the local vicinity of two closely related strains under stress. We chose a Growth Advantage in Stationary Phase (GASP) E scherichia coli strain to address this question because the GASP strain has very similar fitness to the wild-type (WT) strain in the absence of metabolic stress but in the presence of metabolic stress continues to divide and does not enter into stationary phase. We find that under strong antibiotic selection pressure by the fluoroquinolone antibiotic ciprofloxacin in a complex ecology that the GASP strain rapidly evolves in under 20 h missense mutation in gyrA only 2 amino acids removed from the WT strain indicating a convergent solution, yet does not evolve the other 3 mutations of the WT strain. Further the GASP strain evolves a prophage e14 excision which completely inhibits biofilm formation in the mutant strain, revealing the hidden complexity of E. coli evolution to antibiotics as a function of selection pressure. We conclude that there is a cryptic roughness to fitness landscapes in the absence of stress.


Complete assignment of structural genes involved in flavonoid biosynthesis influencing bulb color to individual chromosomes of the shallot (Allium cepa L.).
We analyzed Japanese bunching onion (Allium fistulosum L.) - shallot (Allium cepa L. Aggregatum group) alien chromosome addition lines in order to assign the genes involved in the flavonoid biosynthesis pathway to chromosomes of the shallot. Two complete sets of alien monosomic additions (2n = 2x + 1 = 17) were used for determining the chromosomal locations of several partial sequences of candidate genes, CHS, CHI, F3H, DFR, and ANS via analyses of PCR-based markers. The results of DNA marker analyses showed that the CHS-A, CHS-B, CHI, F3H, DFR, and ANS genes should be assigned to chromosomes 2A, 4A, 3A, 3A, 7A, and 4A, respectively. HPLC analyses of 14 A. fistulosum - shallot multiple alien additions (2n = 2x + 2 - 2x + 7 = 18 - 23) were conducted to identify the anthocyanin compounds produced in the scaly leaves. A direct comparison between the genomic constitution and the anthocyanin compositions of the multiple additions revealed that a 3GT gene for glucosylation of anthocyanidin was located on 4A. Thus, we were able to assign all structural genes involved in flavonoid biosynthesis influencing bulb color to individual chromosomes of A. cepa.


Ex vivo production of platelets from stem cells.
Stem cell technology holds great promise for transfusion medicine, and generation of platelets from stem cells would be transformative. Platelet transfusions are life saving for millions of people and the clinical demand for platelets continues to increase: there is a real need to increase the supply of platelets. Accordingly, there is great interest in the potential of producing platelets from stem cells for clinical use. There has been initial success in ex vivo generation of platelets from stem cells using cord blood stem cells, embryonic stem cells and induced pluripotent stem cells. However, the platelet yields achieved by these strategies have not been sufficient for clinical purposes. This review provides updated information about the current strategies of ex vivo generation of platelets. Megakaryocytopoiesis and platelet generation, along with the importance of genetic determinants of these processes, are reviewed in the context of efforts to generate these products from stem cells. Current challenges and rate-limiting steps in ex vivo platelet generation are discussed, together with strategies to overcome them. While much work remains, great progress has been made, moving ex vivo generation of platelets ever closer to the clinic.


Activated CARD11 accelerates germinal center kinetics, promoting mTORC1 and terminal differentiation.
Activating mutations in the adapter protein CARD11 associated with diffuse large B cell lymphomas (DLBCLs) are predicted to arise during germinal center (GC) responses, leading to inappropriate activation of NF-κB signaling. Here, we modeled the B cell-intrinsic impact of the L251P activating mutation in CARD11 (aCARD11) on the GC response. Global B cell aCARD11 expression led to a modest increase in splenic B cells and a severe reduction in B1 B cell numbers, respectively. Following T cell-dependent immunization, aCARD11 cells exhibited increased rates of GC formation, resolution, and differentiation. Restriction of aCARD11 to GC B cells similarly altered the GC response and B cell differentiation. In this model, aCARD11 promoted dark zone skewing along with increased cycling, AID levels, and class switch recombination. Furthermore, aCard11 GC B cells displayed increased biomass and mTORC1 signaling, suggesting a novel strategy for targeting aCARD11-driven DLBCL. While aCARD11 potently impacts GC responses, the rapid GC contraction suggests it requires collaboration with events that limit terminal differentiation to promote lymphoma.


A chiral lanthanide metal-organic framework for selective sensing of Fe(iii) ions.
A new luminescent lanthanide metal-organic framework was synthesized under hydrothermal conditions by introducing the tetracarboxylic acid ligand (H4bptc) as a chromophore to sensitize the luminescence of the Tb(3+) ion. Interestingly, despite the achiral nature of H4bptc, the resulting framework exhibited rare chiral helical channels. The luminescence studies revealed that this material showed a highly selective fluorescence quenching response to Fe(3+) ions in liquid suspension, which can be conveniently observed by the naked eye under UV light. Thus, this material can be potentially used as a chemosensor for sensing Fe(3+) ions and its selective luminescence quenching response can be explained in terms of the competitive absorption mechanism.


Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus.
Antiviral immunity against a pathogen is mounted upon recognition by the host of virally associated structures. One of these viral 'signatures', double-stranded (ds) RNA, is a replication product of most viruses within infected cells and is sensed by Toll-like receptor 3 (TLR3) and the recently identified cytosolic RNA helicases RIG-I (retinoic acid inducible gene I, also known as Ddx58) and Mda5 (melanoma differentiation-associated gene 5, also known as Ifih1 or Helicard). Both helicases detect dsRNA, and through their protein-interacting CARD domains, relay an undefined signal resulting in the activation of the transcription factors interferon regulatory factor 3 (IRF3) and NF-kappaB. Here we describe Cardif, a new CARD-containing adaptor protein that interacts with RIG-I and recruits IKKalpha, IKKbeta and IKKvarepsilon kinases by means of its C-terminal region, leading to the activation of NF-kappaB and IRF3. Overexpression of Cardif results in interferon-beta and NF-kappaB promoter activation, and knockdown of Cardif by short interfering RNA inhibits RIG-I-dependent antiviral responses. Cardif is targeted and inactivated by NS3-4A, a serine protease from hepatitis C virus known to block interferon-beta production. Cardif thus functions as an adaptor, linking the cytoplasmic dsRNA receptor RIG-I to the initiation of antiviral programmes.


High-Intensity Exercise and Carbohydrate Supplementation do not Alter Plasma Visfatin.
The purpose of the study was to examine the effect of high-intensity exercise and carbohydrate supplementation (CHO) on plasma visfatin. On 2 separate days, 10 sprint-trained males (age = 26.4 ± 5.3 yr; Ht = 1.77 ± 0.03 m; Wt = 78.78 ± 9.10 kg; BF% = 13.96 ± 7.28%) completed 4, 3-min bouts of cycling at 50% mean anaerobic power, with 6 min of rest between bouts. On CHO day, subjects ingested 50g of CHO 30 min before exercise. On control day, subjects ingested a sugar-free drink (CON) 30 min before exercise. Blood was drawn before supplementation, 15 min before exercise, before and after each exercise bout, and 15 and 30 min post exercise. Visfatin, glucose, and insulin were determined. Truncal fat was assessed by dual energy x-ray. Visfatin was not significantly different between treatments (CHO vs CON) at any time point (p = 0.163), and was not significantly altered by exercise (p = 0.692). Insulin [25.65 vs 8.35 mU/l, CHO vs CON, respectively] and glucose [138.57 vs 98.10 mg/dl, CHO vs CON, respectively] were significantly elevated after CHO ingestion and remained elevated throughout the first half of exercise. Baseline visfatin was significantly correlated with truncal fat (r2 = 0.7782, p < 0.05). Visfatin was correlated to truncal fat in sprint-trained males, but was not altered by exercise or CHO supplementation.


The stigma of migraine.
People who have a disease often experience stigma, a socially and culturally embedded process through which individuals experience stereotyping, devaluation, and discrimination. Stigma has great impact on quality of life, behavior, and life chances. We do not know whether or not migraine is stigmatizing. We studied 123 episodic migraine patients, 123 chronic migraine patients, and 62 epilepsy patients in a clinical setting to investigate the extent to which stigma attaches to migraine, using epilepsy as a comparison. We used the stigma scale for chronic illness, a 24-item questionnaire suitable for studying chronic neurologic diseases, and various disease impact measures. Patients with chronic migraine had higher scores (54.0±20.2) on the stigma scale for chronic illness than either episodic migraine (41.7±14.8) or epilepsy patients (44.6±16.3) (p<0.001). Subjects with migraine reported greater inability to work than epilepsy subjects. Stigma correlated most strongly with the mental component score of the short form of the medical outcomes health survey (SF-12), then with ability to work and migraine disability score for chronic and episodic migraine and the Liverpool impact on epilepsy scale for epilepsy. Analysis of covariance showed adjusted scores for the stigma scale for chronic illness were similar for chronic migraine (49.3; 95% confidence interval, 46.2 to 52.4) and epilepsy (46.5; 95% confidence interval, 41.6 to 51.6), and lower for episodic migraine (43.7; 95% confidence interval, 40.9 to 46.6). Ability to work was the strongest predictor of stigma as measured by the stigma scale for chronic illness. In our model, adjusted stigma was similar for chronic migraine and epilepsy, which were greater than for episodic migraine. Stigma correlated most strongly with inability to work, and was greater for chronic migraine than epilepsy or episodic migraine because chronic migraine patients had less ability to work.


Absolute quantitation of glycosylation site occupancy using isotopically labeled standards and LC-MS.
N-linked glycans are required to maintain appropriate biological functions on proteins. Underglycosylation leads to many diseases in plants and animals; therefore, characterizing the extent of glycosylation on proteins is an important step in understanding, diagnosing, and treating diseases. To determine the glycosylation site occupancy, protein N-glycosidase F (PNGase F) is typically used to detach the glycan from the protein, during which the formerly glycosylated asparagine undergoes deamidation to become an aspartic acid. By comparing the abundance of the resulting peptide containing aspartic acid against the one containing non-glycosylated asparagine, the glycosylation site occupancy can be evaluated. However, this approach can give inaccurate results when spontaneous chemical deamidation of the non-glycosylated asparagine occurs. To overcome this limitation, we developed a new method to measure the glycosylation site occupancy that does not rely on converting glycosylated peptides to their deglycosylated forms. Specifically, the overall protein concentration and the non-glycosylated portion of the protein are quantified simultaneously by using heavy isotope-labeled internal standards coupled with LC-MS analysis, and the extent of site occupancy is accurately determined. The efficacy of the method was demonstrated by quantifying the occupancy of a glycosylation site on bovine fetuin. The developed method is the first work that measures the glycosylation site occupancy without using PNGase F, and it can be done in parallel with glycopeptide analysis because the glycan remains intact throughout the workflow.


Early- but not late-adolescent Western diet consumption programs for long-lasting memory impairments in male but not female rats.
Early life Western diet (WD) consumption leads to impaired memory function, particularly for processes mediated by the hippocampus. However, the precise critical developmental window(s) during which WD exposure negatively impacts hippocampal function are unknown. Here, we exposed male and female rats to a WD model involving free access to a variety of high-fat and/or high-sugar food and drink items during either the early-adolescent period (postnatal days [PN] 26-41; WD-EA) or late-adolescent period (PN 41-56; WD-LA). Control (CTL) rats were given healthy standard chow throughout both periods. To evaluate long-lasting memory capacity well beyond the early life WD exposure periods, we performed behavioral assessments after both a short (4 weeks for WD-EA, 2 weeks for WD-LA) and long (12 weeks for WD-EA, 10 weeks for WD-LA) period of healthy diet intervention. Results revealed no differences in body weight or body composition between diet groups, regardless of sex. Following the shorter period of healthy diet intervention, both male and female WD-EA and WD-LA rats showed deficits in hippocampal-dependent memory compared to CTL rats. Following the longer healthy diet intervention period, memory impairments persisted in male WD-EA but not WD-LA rats. In contrast, in female rats the longer healthy diet intervention reversed the initial memory impairments in both WD-EA and WD-LA rats. Collectively, these findings reveal that early-adolescence is a critical period of long-lasting hippocampal vulnerability to dietary insults in male but not female rats, thus highlighting developmental- and sex-specific effects mediating the relationship between the early life nutritional environment and long-term cognitive health.


Nuclear medicine practice in Japan: a report of the sixth nationwide survey in 2007.
The Subcommittee on the Survey of Nuclear Medicine Practice in Japan has performed a nationwide survey of nuclear medicine practice every 5 years since 1982 to provide detailed information on its present status. Questionnaires were sent to all institutions known to the Japan Radioisotope Association to conduct nuclear medicine examinations. The questionnaires addressed the number and kind of nuclear medicine examinations performed as well as the kind and dose of the radiopharmaceuticals used during the month of June 2007. The annual number of total or specific examinations was then estimated. Of the institutions sent questionnaires, 1219 were for in vivo study, 49 for in vitro study, and 212 for positron emission tomography (PET) study. Of these, 92.2% provided answers. A total of 1569 gamma cameras were installed in 1119 institutions, of which 70% were dual-head cameras. The estimated total annual number of in vivo examinations expressed by the number of administered radiopharmaceuticals was 1.41 million, representing a decrease of 11.5% when compared with that of the previous survey (2002). The frequency of study with respect to single-photon emission computed tomography (SPECT) slightly increased to 42.3% from 39.9% in the previous survey. The most frequently performed scintigraphy was bone (38.3%), followed by myocardium (26.2%) and brain perfusion (14.1%). Brain perfusion scintigraphy slightly increased, whereas tumor scintigraphy decreased by one-half when compared with the previous survey. The most commonly used radiopharmaceutical for each scintigraphy was (99m)Tc-HMDP for bone, thallium-201 ((201)Tl)-chloride for myocardium, gallium-67 ((67)Ga)-citrate for tumor, and technetium-99m-ethylcysteinate dimmer ((99m)Tc-ECD) for brain. The number of PET institutes increased from 36 to 212. (18)F-fluorodeoxyglucose ((18)F-FDG)-PET dramatically increased 14.8-fold during the past 5 years. Radionuclide therapy also increased. (131)I therapy for thyroid cancer and hyperthyroidism was conducted yearly in 2373 and 4146 patients, respectively. A total of 13.1 million in vitro radioassays were carried out yearly, the number of which has been decreasing continuously since 1992. It was proved that the content of nuclear medicine practice in Japan has changed considerably in the past 5 years. Namely, (18)F-FDG-PET and radionuclide therapy increased. This report might be useful for understanding the present trends of nuclear medicine practice in Japan.


Acute Respiratory Distress Syndrome Measurement Error. Potential Effect on Clinical Study Results.
Identifying patients with acute respiratory distress syndrome (ARDS) is a recognized challenge. Experts often have only moderate agreement when applying the clinical definition of ARDS to patients. However, no study has fully examined the implications of low reliability measurement of ARDS on clinical studies. To investigate how the degree of variability in ARDS measurement commonly reported in clinical studies affects study power, the accuracy of treatment effect estimates, and the measured strength of risk factor associations. We examined the effect of ARDS measurement error in randomized clinical trials (RCTs) of ARDS-specific treatments and cohort studies using simulations. We varied the reliability of ARDS diagnosis, quantified as the interobserver reliability (κ-statistic) between two reviewers. In RCT simulations, patients identified as having ARDS were enrolled, and when measurement error was present, patients without ARDS could be enrolled. In cohort studies, risk factors as potential predictors were analyzed using reviewer-identified ARDS as the outcome variable. Lower reliability measurement of ARDS during patient enrollment in RCTs seriously degraded study power. Holding effect size constant, the sample size necessary to attain adequate statistical power increased by more than 50% as reliability declined, although the result was sensitive to ARDS prevalence. In a 1,400-patient clinical trial, the sample size necessary to maintain similar statistical power increased to over 1,900 when reliability declined from perfect to substantial (κ = 0.72). Lower reliability measurement diminished the apparent effectiveness of an ARDS-specific treatment from a 15.2% (95% confidence interval, 9.4-20.9%) absolute risk reduction in mortality to 10.9% (95% confidence interval, 4.7-16.2%) when reliability declined to moderate (κ = 0.51). In cohort studies, the effect on risk factor associations was similar. ARDS measurement error can seriously degrade statistical power and effect size estimates of clinical studies. The reliability of ARDS measurement warrants careful attention in future ARDS clinical studies.


Do primary care physicians require training in core clinical dermatology? A cross sectional needs assessment study from Western India.
Primary care physicians play a crucial role in managing patients with common skin disorders who form around one-third of outpatient attendees. This study aimed to assess the need for dermatology training among primary care physicians by assessing their knowledge, self-perception of ability to diagnose and manage skin disorders, and their difficulties in managing these patients. A descriptive, cross-sectional, needs assessment study was done among primary care physicians (n = 61) of rural (n = 34) and urban (n = 27) areas of Vadodara district. A pre-validated semi-structured questionnaire (for self-rating of the ability to diagnose/manage skin disorders and difficulties faced in managing patients) was used along with a photo-quiz (for knowledge assessment) while approaching primary care physicians during their monthly review meeting with prior permission. Data were analyzed by Epi InfoTM software and manual content analysis. The mean score on the photo-quiz was 4.1/10. Forty-three (70.5%) participants rated their ability to diagnose/manage skin disorders as 'average' on a five-point Likert scale. Various difficulties (n = 89) narrated by participants were related to their knowledge/skill, disease factors, patients and administrative aspects. Three-fourths of the participants managed difficulties by referring patients to dermatologists. One-third suggested conducting training in common skin disorders. The study population included primary care physicians from the government healthcare setup only. Knowledge assessment was done using a short ten-item photo-quiz instead of actual patients. Primary care physicians had poor knowledge of skin disorders, and a majority overrated their own ability for clinical management of these disorders. Most common difficulties faced were related to clinical management. There is a need for training on common skin disorders.


Interleukin-4 is a potent mitogen for capillary endothelium.
Interleukin-4 (IL-4) is a mitogen for both microvascular (human adrenal capillary, HACE) and large vessel (human umbilical vein, HUVEC) endothelial cells. Comparison of growth promotion by IL-4 to that by the potent endothelial mitogen fibroblast growth factor (FGF) showed the activity of IL-4 on HACE cells to be strong (50% of that with FGF) but on HUVEC's weak (12% of that with FGF). Growth stimulation was characterised by both 3H-thymidine incorporation and by cell number, and was maximal at 1 nM IL-4. The presence of IL-4 receptors on HACE cells and HUVEC's was confirmed by specific binding of radioiodinated IL-4. Scatchard analysis confirmed a single high affinity binding receptor on both HACE cells (Kd = 80 pM, 358 receptors/cell) and HUVEC's (Kd = 88 pM, 2,580 receptors/cell). Potent activity on capillary as opposed to large vessel endothelium places IL-4 in a unique position amongst endothelial mitogens.


Drinking water residence time in distribution networks and emergency department visits for gastrointestinal illness in Metro Atlanta, Georgia.
We examined whether the average water residence time, the time it takes water to travel from the treatment plant to the user, for a zip code was related to the proportion of emergency department (ED) visits for gastrointestinal (GI) illness among residents of that zip code. Individual-level ED data were collected from all hospitals located in the five-county metro Atlanta area from 1993 to 2004. Two of the largest water utilities in the area, together serving 1.7 million people, were considered. People served by these utilities had almost 3 million total ED visits, 164,937 of them for GI illness. The relationship between water residence time and risk for GI illness was assessed using logistic regression, controlling for potential confounding factors, including patient age and markers of socioeconomic status (SES). We observed a modestly increased risk for GI illness for residents of zip codes with the longest water residence times compared with intermediate residence times (odds ratio (OR) for Utility 1 = 1.07, 95% confidence interval (CI) = 1.03, 1.10; OR for Utility 2 = 1.05, 95% CI = 1.02, 1.08). The results suggest that drinking water contamination in the distribution system may contribute to the burden of endemic GI illness.


Fast track randomized controlled trial to prevent externalizing psychiatric disorders: findings from grades 3 to 9.
This study tests the efficacy of the Fast Track Program in preventing antisocial behavior and psychiatric disorders among groups varying in initial risk. Schools within four sites (Durham, NC; Nashville, TN; Seattle, WA; and rural central Pennsylvania) were selected as high-risk institutions based on neighborhood crime and poverty levels. After screening 9,594 kindergarteners in these schools, 891 highest risk and moderate-risk children (69% male and 51% African American) were randomly assigned by matched sets of schools to intervention or control conditions. The 10-year intervention (begun in 1991 with three yearly cohorts) included parent behavior-management training, child social-cognitive skills training, reading tutoring, home visiting, mentoring, and a universal classroom curriculum. Outcomes included criterion counts and psychiatric diagnoses after grades 3, 6, and 9 for conduct disorder, oppositional defiant disorder, attention-deficit/hyperactivity disorder, any externalizing disorder, and self-reported antisocial behavior. Grade 9 outcomes were assessed between 2000 and 2003, depending upon cohort. Significant interaction effects between intervention and initial risk level were found at each age but most strongly after grade 9. Assignment to intervention had a significant positive effect in lowering criterion count scores and diagnoses for conduct disorder, attention-deficit/hyperactivity disorder, and any externalizing disorder, and lowering antisocial behavior scores, but only among those at highest risk initially. Prevention of serious antisocial behavior can be efficacious across sex, ethnicity, and urban/rural residence, but screening is essential.


[Scientific production and licensing of medical schools in Peru].
The new university law 30220 of 2014 introduced the mandatory institutional licensing of all Peruvian universities by the National Superintendence of Higher University Education (SUNEDU, in Spanish). The first undergraduate program to go through this process will be medicine. The licensing of medical programs is necessary to ensure that the conditions in which the program is taught in Peru are adequate, with a high probability of closing some medical schools. Once a medical school has demonstrated that it meets the basic conditions of quality, a qualitative and quantitative evaluation is carried out that includes three criteria: scientific production in the Web of Science, impact measured through the H index, and results of the national medical exam, to determine the years of licensing. This article evaluates the quantitative indicators linked to research using Web of Science and Scopus, in addition to making technical and methodological revisions of them. Suggestions for the other indicators are also covered by this article.


Effect of Electronic Cigarette Use on the Urge-to-Cough Sensation.
Electronic cigarettes (e-cigs) have attained common usage worldwide, yet knowledge of their physiological effects remains minimal. The aim of this study was to evaluate the effect of a single exposure to e-cig vapor on the urge-to-cough (UTC) threshold and C5. Seventeen healthy nonsmokers underwent C5 measurement employing capsaicin cough challenge at baseline, 15 minutes, and 24 hours after e-cig exposure (30 puffs 30 seconds apart). The endpoint of cough challenge is C5, the concentration of capsaicin inducing five or more coughs. The UTC threshold (Cu) is defined as the lowest concentration of capsaicin inducing UTC without an associated motor cough. The Cu and C5 were significantly inhibited (Cu and C5 increased) 15 minutes after e-cig use. Mean log Cu rose from -0.035±0.08 (SEM) to 0.21±0.12 (P = .005). Mean log C5 increased from 0.60±0.11 to 0.92±0.16 (P = .001). By 24 hours after e-cig exposure, Cu and C5 had returned to baseline levels. A single session of e-cig use, approximating nicotine exposure of one tobacco cigarette, induces significant inhibition of the Cu, as well as C5. Previous studies implicate nicotine as the agent responsible for suppression of C5, and we hypothesize a similar role for nicotine in the suppression of the Cu. Given our observation of the effect of a single e-cig exposure, studies of the respiratory physiologic implications of repeated or chronic e-cig use are warranted. This is the first study to demonstrate that a single exposure to an e-cig significantly inhibits the Cu as measured by capsaicin cough challenge testing. These findings add to the growing body of evidence that e-cig vapor is not a physiologically benign substance, and support further investigation of the effects of repeated or chronic use of e-cigs on cough sensitivity and other respiratory parameters.


Footwear characteristics and foot problems in older people.
Foot problems are common in older people, however the contribution of incorrectly fitting footwear and heel elevation to the development of foot pain and deformity has not been fully evaluated. To examine the relationship between footwear characteristics and the prevalence of common forefoot problems in older people. Presence of foot pain and deformity were identified in 176 people (56 men and 120 women) aged 62-96 years (mean 80.09, SD 6.42) using a questionnaire and clinical assessment. Shoe fit was determined by comparing length, width and area measurements of shoes with foot measurements. Past and present use of high heels in women was documented, and heel elevation of footwear was measured. Most subjects wore shoes narrower than their feet. Women wore shoes that were shorter, narrower and had a reduced total area compared to their feet than men. Wearing shoes substantially narrower than the foot was associated with corns on the toes, hallux valgus deformity and foot pain, whereas wearing shoes shorter than the foot was associated with lesser toe deformity. Wearing shoes with heel elevation greater than 25 mm was associated with hallux valgus and plantar calluses in women. Incorrectly fitting footwear is common in older people and is strongly associated with forefoot pathology and foot pain. These findings highlight the need for footwear assessment in the management of foot problems in older people.


Oral sex practices among men who have sex with men and transgender women at risk for and living with HIV in Nigeria.
Men who have sex with men (MSM) and transgender women (TGW) are at risk for sexually transmitted infections (STIs), including those of the oropharynx. We estimated the prevalence and factors associated with oral sex practices and characterized oropharyngeal STIs among a cohort of MSM and TGW in Nigeria. From 2013 to 2018, TRUST/RV368 recruited MSM and TGW into HIV/STI diagnosis and treatment at community-based clinics in Nigeria. Participants who completed HIV testing and oral sex questions at enrollment were selected. Cross-sectional analyses with bivariate and multivariable logistic regression models estimated odds ratios (ORs) and 95% confidence intervals (CIs). Oropharyngeal swab testing for Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) began in 2014 and for those with diagnostic results at enrollment, the unadjusted association of oral sex practices with oropharyngeal STIs was conducted. A total of 1342 participants had a median age of 25 years (interquartile range: 22-29), 58% were living with HIV, and 69% reported oral sex practices. Factors associated with increased odds of engaging in oral sex included living with HIV (adjusted [a]OR: 1.4, 95% CI: 1.1-1.8), self-identifying as a woman (aOR:1.8, 95% CI: 1.1-2.8), mobile phone ownership (aOR:2.3, 95% CI: 1.3-3.9), receptive anal sex (aOR:1.7, 95% CI:1.3-2.3) and multiple male sexual partners (2 to 4 vs. ≤1, aOR:1.5, 95% CI: 1.0-2.2; 5+ vs ≤1, aOR:2.9, 95% CI:1.9-4.3). Oropharyngeal STI prevalence was 7% (52/752) and higher among those who engaged in oral sex compared to those who did not (unadjusted OR: 2.5, 95% CI:1.2-5.3). Oral sex was common and associated with an increased odds of oropharyngeal STIs among MSM and TGW from Nigeria. In the absence of screening and treatment guidelines, condoms continue to be the mainstay for oral STI prevention. A pre-exposure prophylaxis for bacterial STIs would complement current prevention strategies to curb transmission.


Variation in regional and landscape effects on occupancy of temperate bats in the southeastern U.S.
Habitat loss, wind energy development, and the disease white-nose syndrome are major threats contributing to declines in bat populations in North America. In the southeastern US in particular, the recent arrival of white-nose syndrome and changes in landscape composition and configuration have driven shifts in bat species populations and distributions. Effective management strategies which address these large-scale, community-level threats require landscape-scale analyses. Our objective was to model the relationship between ecoregional and landscape factors and occupancy by all bat species in South Carolina, USA, during summer. We conducted acoustic surveys from mid-May through July 2015 and 2016 and evaluated temporally dynamic occupancy models for eight bat species or species groups at the 100 km2 level. We found significant effects of landscape factors such as ecoregion and forest edge density for three species, but habitat condition effects were not statistically significant for five other species. Thus, for some species, site-use analyses may be more appropriate than larger scale occupancy analyses. However, our occupancy predictions generally matched statewide historical distributions for all species, suggesting our approach could be useful for monitoring landscape-level trends in bat species. Thus, while our scale of study was likely too coarse for assessing fine-scale habitat associations for all bat species, our findings can improve future monitoring efforts, inform conservation priorities, and guide subsequent landscape-scale studies for bat species and community-level responses to global change.


Dynamic modeling of nutrient use and individual requirements of lactating sows1.
Nutrient requirements of sows during lactation are related mainly to their milk yield and feed intake, and vary greatly among individuals. In practice, nutrient requirements are generally determined at the population level based on average performance. The objective of the present modeling approach was to explore the variability in nutrient requirements among sows by combining current knowledge about nutrient use with on-farm data available on sows at farrowing [parity, BW, backfat thickness (BT)] and their individual performance (litter size, litter average daily gain, daily sow feed intake) to estimate nutrient requirements. The approach was tested on a database of 1,450 lactations from 2 farms. The effects of farm (A, B), week of lactation (W1: week 1, W2: week 2, W3+: week 3 and beyond), and parity (P1: 1, P2: 2, P3+: 3 and beyond) on sow performance and their nutrient requirements were evaluated. The mean daily ME requirement was strongly correlated with litter growth (R2 = 0.95; P < 0.001) and varied slightly according to sow BW, which influenced the maintenance cost. The mean daily standardized ileal digestible (SID) lysine requirement was influenced by farm, week of lactation, and parity. Variability in SID lysine requirement per kg feed was related mainly to feed intake (R2 = 0.51; P < 0.001) and, to a smaller extent, litter growth (R2 = 0.27; P < 0.001). It was lowest in W1 (7.0 g/kg), greatest in W2 (7.9 g/kg), and intermediate in W3+ (7.5 g/kg; P < 0.001) because milk production increased faster than feed intake capacity did. It was lower for P3+ (6.7 g/kg) and P2 sows (7.3 g/kg) than P1 sows (8.3 g/kg) due to the greater feed intake of multiparous sows. The SID lysine requirement per kg of feed was met for 80% of sows when supplies were 112 and 120% of the mean population requirement on farm A and B, respectively, indicating higher variability in requirements on farm B. Other amino acid and mineral requirements were influenced in the same way as SID lysine. The present modeling approach allows to capture individual variability in the performance of sows and litters according to farm, stage of lactation, and parity. It is an initial step in the development of new types of models able to process historical farm data (e.g., for ex post assessment of nutrient requirements) and real-time data (e.g., to control precision feeding).


Adaptive noise reduction method for DSPI fringes based on bi-dimensional ensemble empirical mode decomposition.
Digital speckle pattern interferometry (DSPI) fringes contain low spatial information degraded with speckle noise and background intensity. The denoising technique proposed recently based on bi-dimensional empirical mode decomposition (BEMD) could implement noise reduction adaptively. However, the major drawback of BEMD, called mode mixing, has affected its practical application. With noise-assisted data analysis (NADA) method, bi-dimensional ensemble empirical mode decomposition (BEEMD) was proposed, which has solved the problem of mode mixing. The denoising approach based on BEEMD will be presented, compared with other classic denoising methods and evaluated both qualitatively and quantitatively using computer-simulated and experimental DSPI fringes.


hs-Troponin I Followed by CT Angiography Improves Acute Coronary Syndrome Risk Stratification Accuracy and Work-Up in Acute Chest Pain Patients: Results From ROMICAT II Trial.
This study compared diagnostic accuracy of conventional troponin/traditional coronary artery disease (CAD) assessment and highly sensitive troponin (hsTn) I/advanced CAD assessment for acute coronary syndrome (ACS) during the index hospitalization. hsTnI and advanced assessment of CAD using coronary computed tomography angiography (CTA) are promising candidates to improve the accuracy of emergency department evaluation of patients with suspected ACS. We performed an observational cohort study in patients with suspected ACS enrolled in the ROMICAT II (Rule Out Myocardial Infarction/Ischemia using Computer Assisted Tomography) trial and randomized to coronary CTA who also had hsTnI measurement at the time of the emergency department presentation. We assessed coronary CTA for traditional (no CAD, nonobstructive CAD, ≥50% stenosis) and advanced features of CAD (≥50% stenosis, high-risk plaque features: positive remodeling, low <30-Hounsfield units plaque, napkin-ring sign, spotty calcium). Of 160 patients (mean age: 53 ± 8 years, 40% women) 10.6% were diagnosed with ACS. The ACS rate in patients with hsTnI below the limit of detection (n = 9, 5.6%), intermediate (n = 139, 86.9%), and above the 99th percentile (n = 12, 7.5%) was 0%, 8.6%, and 58.3%, respectively. Absence of ≥50% stenosis and high-risk plaque ruled out ACS in patients with intermediate hsTnI (n = 87, 54.4%; ACS rate 0%), whereas patients with both ≥50% stenosis and high-risk plaque were at high risk (n = 13, 8.1%; ACS rate 69.2%) and patients with either ≥50% stenosis or high-risk plaque were at intermediate risk for ACS (n = 39, 24.4%; ACS rate 7.7%). hsTnI/advanced coronary CTA assessment significantly improved the diagnostic accuracy for ACS as compared to conventional troponin/traditional coronary CTA (area under the curve 0.84, 95% confidence interval [CI]: 0.80 to .88 vs. 0.74, 95% CI: 0.70 to 0.78; p < 0.001). hsTnI at the time of presentation followed by early advanced coronary CTA assessment improves the risk stratification and diagnostic accuracy for ACS as compared to conventional troponin and traditional coronary CTA assessment. (Multicenter Study to Rule Out Myocardial Infarction/Ischemia by Cardiac Computed Tomography [ROMICAT-II]; NCT01084239).


Estimation of glomerular filtration rate via 2- and 4-sample plasma clearance of iohexol and creatinine in clinically normal cats.
To compare 2 methods for estimation of glomerular filtration rate (GFR), study the effects of age and body size on GFR estimates, and provide a reference range for estimated GFR in clinically normal cats. 57 cats. In each cat, GFR was estimated via plasma clearance of iohexol and creatinine. Results of a 1-compartmental model (CL1comp) were calibrated to a trapezoidal method estimate (CLtrap) by use of a correction formula applicable to dogs or humans and standardized to body weight; for iohexol clearance, data were also standardized to extracellular fluid volume (ECFV). For all 57 cats, method comparison was performed via agreement analysis. Reference ranges for GFR derived by the different methods were established by use of data from a subset of 51 cats after exclusion of 6 cats that were azotemic, Birman, or both. In 57 cats, mean CLtrap of creatinine was 0.29 mL/min/kg (13%) higher than CLtrap of iohexol. In 51 nonazotemic cats, mean CLtrap was 2.26 mL/min/kg for iohexol (reference range, 1.02 to 3.50 mL/min/kg) and 2.55 mL/min/kg for creatinine (reference range, 1.27 to 3.83 mL/min/kg). Values of GFR/kg or GFR standardized to liters of ECFV did not decrease with increasing age. A negative linear relationship was detected between body weight and estimated GFR/kg or GFR standardized to liters of ECFV. Reference ranges for estimated GFR via plasma clearance of iohexol and creatinine should facilitate early detection of impaired renal function in cats, although body weight should be taken into account.


Regulation of Pseudomonas aeruginosa internalization after contact lens wear in vivo and in serum-free culture by ocular surface cells.
To determine the effects of contact lenses (CLs) and Pseudomonas aeruginosa (PA) infection on localization of cystic fibrosis transmembrane conductance regulator (CFTR) on corneal surface epithelial cells and the association between lipid raft formation and CFTR in mediating PA binding and internalization in ocular surface epithelium. CFTR immunolocalization was evaluated in vivo in rabbit corneal-conjunctival epithelium (with/without CL wear) before and after PA exposure and in serum-free human corneal epithelial cell culture (hTCEpi). Lipid raft formation was visualized with Alexa555-conjugated cholera toxin beta-subunit. Lipid raft involvement in PA internalization was assayed in vivo by gentamicin survival assays after topical filipin pretreatment. Involvement of CFTR in PA binding and internalization was evaluated by blockade with CFTR peptides or LPS. CL wear in vivo enhanced anti-CFTR staining, but CFTR localization did not correlate with the PA binding by ocular surface cells. Conjunctival epithelial cells stained for CFTR but did not bind or internalize PA. Corneal epithelial cells in vivo did not stain for CFTR unless challenged by contact lens-induced hypoxia. PA internalization by hTCEpi was significantly inhibited by LPS (P < 0.01), but not by CFTR peptides. Remarkably, normal conjunctival epithelial cells showed lipid raft formation and CFTR staining but did not bind PA. Inhibition of raft formation by filipin blocked PA internalization in vivo after CL wear. CFTR is not the predominant receptor for ocular surface PA infection, and after hypoxic CL challenge, neither lipid rafts nor CFTR localization alone predicts PA binding; however, lipid rafts are critical to CL-mediated PA internalization.


The dissolution and solid-state behaviours of coground ibuprofen-glucosamine HCl.
The cogrinding technique is one of most effective methods for improving the dissolution of poorly water-soluble drugs and it is superior to other approaches from an economical as well as an environmental standpoint, as the technique does not require any toxic organic solvents. Present work explores the role of d-glucosamine HCl (GL) as a potential excipient to improve dissolution of a low melting point drug, ibuprofen (Ibu), using physical mixtures and coground formulations. The dissolution of the poorly soluble drug has been improved by changing the ratio of Ibu:GL and also grinding time. The results also showed that although GL can enhance the solubility of Ibu, it also reduces pH around the Ibu particles which led to poor dissolution performance when the concentration of GL is high. The effect of GL on the solubility of Ibu could be misleading if the pH of the final solution was not measured. Grinding reduced the particle size of GL significantly but in case of Ibu it was less effective. Solid state analysis (XRPD, DSC, and FT-IR) showed that ibuprofen is stable under grinding conditions, but the presence of high concentration of GL in samples subjected to high grinding times caused changes in FT-IR spectrum of Ibu which could be due to intermolecular hydrogen bond or esterification between the carboxylic acid group in the ibuprofen and hydroxyl group in the GL.


Recovering population parameters from a single gene genealogy: an unbiased estimator of the growth rate.
We show that the number of lineages ancestral to a sample, as a function of time back into the past, which we call the number of lineages as a function of time (NLFT), is a nearly deterministic property of large-sample gene genealogies. We obtain analytic expressions for the NLFT for both constant-sized and exponentially growing populations. The low level of stochastic variation associated with the NLFT of a large sample suggests using the NLFT to make estimates of population parameters. Based on this, we develop a new computational method of inferring the size and growth rate of a population from a large sample of DNA sequences at a single locus. We apply our method first to a sample of 1,212 mitochondrial DNA (mtDNA) sequences from China, confirming a pattern of recent population growth previously identified using other techniques, but with much smaller confidence intervals for past population sizes due to the low variation of the NLFT. We further analyze a set of 63 mtDNA sequences from blue whales (BWs), concluding that the population grew in the past. This calls for reevaluation of previous studies that were based on the assumption that the BW population was fixed.


Phosphorylation of Exo1 modulates homologous recombination repair of DNA double-strand breaks.
DNA double-strand break (DSB) repair via the homologous recombination pathway is a multi-stage process, which results in repair of the DSB without loss of genetic information or fidelity. One essential step in this process is the generation of extended single-stranded DNA (ssDNA) regions at the break site. This ssDNA serves to induce cell cycle checkpoints and is required for Rad51 mediated strand invasion of the sister chromatid. Here, we show that human Exonuclease 1 (Exo1) is required for the normal repair of DSBs by HR. Cells depleted of Exo1 show chromosomal instability and hypersensitivity to ionising radiation (IR) exposure. We find that Exo1 accumulates rapidly at DSBs and is required for the recruitment of RPA and Rad51 to sites of DSBs, suggesting a role for Exo1 in ssDNA generation. Interestingly, the phosphorylation of Exo1 by ATM appears to regulate the activity of Exo1 following resection, allowing optimal Rad51 loading and the completion of HR repair. These data establish a role for Exo1 in resection of DSBs in human cells, highlighting the critical requirement of Exo1 for DSB repair via HR and thus the maintenance of genomic stability.


Genetic diversity, population structure, effective population size and demographic history of the Finnish wolf population.
The Finnish wolf population (Canis lupus) was sampled during three different periods (1996-1998, 1999-2001 and 2002-2004), and 118 individuals were genotyped with 10 microsatellite markers. Large genetic variation was found in the population despite a recent demographic bottleneck. No spatial population subdivision was found even though a significant negative relationship between genetic relatedness and geographic distance suggested isolation by distance. Very few individuals did not belong to the local wolf population as determined by assignment analyses, suggesting a low level of immigration in the population. We used the temporal approach and several statistical methods to estimate the variance effective size of the population. All methods gave similar estimates of effective population size, approximately 40 wolves. These estimates were slightly larger than the estimated census size of breeding individuals. A Bayesian model based on Markov chain Monte Carlo simulations indicated strong evidence for a long-term population decline. These results suggest that the contemporary wolf population size is roughly 8% of its historical size, and that the population decline dates back to late 19th century or early 20th century. Despite an increase of over 50% in the census size of the population during the whole study period, there was only weak evidence that the effective population size during the last period was higher than during the first. This may be caused by increased inbreeding, diminished dispersal within the population, and decreased immigration to the population during the last study period.


Nickel, cobalt, zinc and copper levels in brown trout (Salmo trutta) from the river Otra, southern Norway.
The Flåt Nickel mine at Evje in southern Norway was mined extensively from 1914 to 1945 with little regard for any potential environmental effect. Much of the ore extracted was smelted at a site adjacent to the river Otra south of Evje. Recent studies have revealed heavy metal pollution in the land surrounding the smelter and in water draining from the mine leading to concern for the aquatic ecosystem in the river Otra. Brown trout were sampled from an uncontaminated lake 9 km upstream from the smelter, from the base of the Oddebekken (a tributary draining the mine water into the Otra), from sites immediately upstream and down stream of the smelter and from a site 4 km down stream from the smelter. Fish from sites adjacent to the smelter and the base of the Oddebekken were smaller than those from the lake and down stream site. Concentrations of the metals were highest in fish sampled where the mine water entered the Otra and gradually decreased in fish further down the river. Fish from the uncontaminated lake had the lowest level of metals.


Inhibition of human glutathione reductase by the nitrosourea drugs 1,3-bis(2-chloroethyl)-1-nitrosourea and 1-(2-chloroethyl)-3-(2-hydroxyethyl)-1-nitrosourea. A crystallographic analysis.
Glutathione reductase from human erythrocytes was inhibited by incubation with the drugs 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and 1-(2-chloroethyl)-3-(2-hydroxyethyl)-1-nitrosourea (HeCNU) under quasi-physiological conditions. For reference purposes, iodoacetamide was used for inactivating alkylation of the enzyme. In each case the modified glutathione reductase was crystallized and its structure determined. These analyses showed that in all experiments the enzyme had reacted at the distal sulfur, that is at the thiol of Cys-58, and virtually nowhere else in the visible structure. The electron density of the HeCNU derivative at 0.3 nm resolution is consistent with a 2-hydroxyethyl group. This alkyl moiety has recently been identified by chemical analysis [Schirmer, R. H., Schöllhammer, T., Eisenbrand, G. and Krauth-Siegel, R. L. (1987) Free Radical Res. Commun. 3, 3-12]. The 0.2 nm resolution electron-density map of the BCNU-derivatized enzyme cannot be explained by a 2-hydroxyethyl group. Instead the modification appears as a carbamoyl moiety containing at least five non-hydrogen atoms. In this derivative the distal cysteine is forced into an unusual conformation.


Differential expression and novel permeability properties of three aquaporin 8 paralogs from seawater-challenged Atlantic salmon smolts.
Aquaporins may facilitate transepithelial water absorption in the intestine of seawater (SW)-acclimated fish. Here we have characterized three full-length aqp8 paralogs from Atlantic salmon (Salmo salar). Bayesian inference revealed that each paralog is a representative of the three major classes of aqp8aa, aqp8ab and aqp8b genes found in other teleosts. The permeability properties were studied by heterologous expression in Xenopus laevis oocytes, and the expression levels examined by qPCR, immunofluorescence and immunoelectron microscopy, and immunoblotting of membrane fractions from intestines of SW-challenged smolts. All three Aqp8 paralogs were permeable to water and urea, whereas Aqp8ab and -8b were, surprisingly, also permeable to glycerol. The mRNA tissue distribution of each paralog was distinct, although some tissues such as the intestine showed redundant expression of more than one paralog. Immunofluorescence microscopy localized Aqp8aa(1+2) to intracellular compartments of the liver and intestine, and Aqp8ab and Aqp8b to apical plasma membrane domains of the intestinal epithelium, with Aqp8b also in goblet cells. In a control experiment with rainbow trout, immunoelectron microscopy confirmed abundant labeling of Aqp8ab and -8b at apical plasma membranes of enterocytes in the middle intestine and also in subapical vesicular structures. During SW challenge, Aqp8ab showed significantly increased levels of protein expression in plasma-membrane-enriched fractions of the intestine. These data indicate that the Atlantic salmon Aqp8 paralogs have neofunctionalized on a transcriptional as well as a functional level, and that Aqp8ab may play a central role in the intestinal transcellular uptake of water during SW acclimation.


The novel EpCAM-targeting monoclonal antibody 3-17I linked to saporin is highly cytotoxic after photochemical internalization in breast, pancreas and colon cancer cell lines.
The epithelial cell adhesion molecule (EpCAM) is expressed by a wide range of human carcinomas, making it an attractive diagnostic and therapeutic target in oncology. Its recent identification on cancer stem cells has raised further interest in its use for tumor targeting and therapy. Here, we present the characterization and therapeutic potential of 3-17I, a novel human EpCAM-targeting monoclonal antibody. Strong reaction of 3-17I was observed in all lung, colon, and breast human tumor biopsies evaluated. By flow cytometry and confocal fluorescence microscopy, we demonstrate that 3-17I specifically targets EpCAM-positive cell lines. We also show evidence for mAb-sequestration in endo-/lysosomes, suggesting internalization of 3-17I by receptor-mediated endocytosis. The ribosomal-inactivating toxin saporin was linked to 3-17I, creating the per se non-toxic immunotoxin 3-17I-saporin, a promising candidate for the drug delivery technology photochemical internalization (PCI). PCI is based on a light-controlled destruction of endolysosomal membranes and subsequent cytosolic release of the sequestered payload upon light exposure. EpCAM-positive human cancer cell lines MCF7 (breast), BxPC-3 (pancreas), WiDr (colon), and the EpCAM-negative COLO320DM (colon), were treated with 3-17I-saporin in combination with the clinically relevant photosensitizer TPCS2a (Amphinex), followed by exposure to light. No cytotoxicity was observed after treatment with 3-17I-saporin without light exposure. However, cell viability, proliferation and colony-forming capacity was strongly reduced in a light-dependent manner after PCI of 3-17I. Our results show that 3-17I is an excellent candidate for diagnosis of EpCAM-positive tumors and for development of clinically relevant antibody-drug conjugates, using PCI for the treatment of localized tumors.


What is the learning curve for carotid artery stenting with neuroprotection? Analysis of 200 consecutive cases at an academic institution.
Carotid artery stenting (CAS) has emerged as an acceptable treatment alternative in high-risk patients with carotid stenosis. This study assessed the effect of the learning curve on treatment complications and clinical outcome of CAS. Clinical variables and treatment outcomes of 200 consecutive CAS procedures in 182 patients (mean age, 72 years) with carotid stenosis of 70% or greater during a 40-month period were analyzed. Technical success, periprocedural complications, and treatment outcomes were compared in four sequential groups (group I, II, III, and IV) of 50 consecutive interventions. Treatment indications and relevant risk factors were similar among the four groups. The overall technical success was 98%, and the combined 30-day stroke and death rates was 2.5%. An increase in the technical success rate was noted in the latter three groups compared with group I (P < .05). Total procedural time and contrast volume were significantly higher in group I compared with the latter three groups (P < .05). The intraoperative anticoagulation regimen was changed from an intravenous heparin combination to bivalirudin after the first 54 cases, resulting in reduced bleeding complications in groups III and IV (P = .03) compared with group I. The 30-day stroke and death rate in groups I and II was 8% and 2%, which was reduced significantly to 0% in groups III and IV (P < .05). A Cox regression model identified procedural volume (P = .03) as a predictor of a reduced complication rate. Carotid artery stenting with neuroprotection can provide excellent treatment outcome. Our experience demonstrates a procedural-associated learning curve, as evidenced by the reduced procedural-related complications, fluoroscopic time, and contrast volume that occurred with an increase in physician experience. The procedural success is also enhanced partly by endovascular device refinement and improved anticoagulation regimen. Successful outcome of CAS can be achieved once physicians overcome the initial procedural-related learning curve.


Finnish and Swedish nursing students' experiences of their first clinical practice placement--a qualitative study.
Nursing is a practice-based discipline. Clinical practice settings are important in preparing undergraduate nursing students for the role of registered nurse. The aim of this Nordic qualitative study is to illuminate first year undergraduate nursing students' experiences of clinical practice during their first clinical placement, with a focus on preception, reflection, and the link between theory and practice. Data were collected by focus group interviews with 22 nursing students, and analyzed with qualitative content analysis. Positive experiences included stimulating and visible preceptors, a permissive atmosphere, and reflection as a matter of course. Negative experiences were related to feelings of abandonment and powerlessness when preceptors were invisible and the atmosphere at the ward was non-permissive. The implementation of research-based knowledge was insufficient. A permissive atmosphere and visible preceptors are crucial if learning is to be maximized. Consequently, it is important to set aside time for preceptors to be more visible and to make the atmosphere at the clinical placement more permissive. The student must have the opportunity to combine scientific knowledge with evidence-based knowledge in order to develop nursing actions.


Construction of the first genetic linkage map of Japanese gentian (Gentianaceae).
Japanese gentians (Gentiana triflora and Gentiana scabra) are amongst the most popular floricultural plants in Japan. However, genomic resources for Japanese gentians have not yet been developed, mainly because of the heterozygous genome structure conserved by outcrossing, the long juvenile period, and limited knowledge about the inheritance of important traits. In this study, we developed a genetic linkage map to improve breeding programs of Japanese gentians. Enriched simple sequence repeat (SSR) libraries from a G. triflora double haploid line yielded almost 20,000 clones using 454 pyrosequencing technology, 6.7% of which could be used to design SSR markers. To increase the number of molecular markers, we identified three putative long terminal repeat (LTR) sequences using the recently developed inter-primer binding site (iPBS) method. We also developed retrotransposon microsatellite amplified polymorphism (REMAP) markers combining retrotransposon and inter-simple sequence repeat (ISSR) markers. In addition to SSR and REMAP markers, modified amplified fragment length polymorphism (AFLP) and random amplification polymorphic DNA (RAPD) markers were developed. Using 93 BC1 progeny from G. scabra backcrossed with a G. triflora double haploid line, 19 linkage groups were constructed with a total of 263 markers (97 SSR, 97 AFLP, 39 RAPD, and 30 REMAP markers). One phenotypic trait (stem color) and 10 functional markers related to genes controlling flower color, flowering time and cold tolerance were assigned to the linkage map, confirming its utility. This is the first reported genetic linkage map for Japanese gentians and for any species belonging to the family Gentianaceae. As demonstrated by mapping of functional markers and the stem color trait, our results will help to explain the genetic basis of agronomic important traits, and will be useful for marker-assisted selection in gentian breeding programs. Our map will also be an important resource for further genetic analyses such as mapping of quantitative trait loci and map-based cloning of genes in this species.


NK Cells Preferentially Target Tumor Cells with a Cancer Stem Cell Phenotype.
Increasing evidence supports the hypothesis that cancer stem cells (CSCs) are resistant to antiproliferative therapies, able to repopulate tumor bulk, and seed metastasis. NK cells are able to target stem cells as shown by their ability to reject allogeneic hematopoietic stem cells but not solid tissue grafts. Using multiple preclinical models, including NK coculture (autologous and allogeneic) with multiple human cancer cell lines and dissociated primary cancer specimens and NK transfer in NSG mice harboring orthotopic pancreatic cancer xenografts, we assessed CSC viability, CSC frequency, expression of death receptor ligands, and tumor burden. We demonstrate that activated NK cells are capable of preferentially killing CSCs identified by multiple CSC markers (CD24(+)/CD44(+), CD133(+), and aldehyde dehydrogenase(bright)) from a wide variety of human cancer cell lines in vitro and dissociated primary cancer specimens ex vivo. We observed comparable effector function of allogeneic and autologous NK cells. We also observed preferential upregulation of NK activation ligands MICA/B, Fas, and DR5 on CSCs. Blocking studies further implicated an NKG2D-dependent mechanism for NK killing of CSCs. Treatment of orthotopic human pancreatic cancer tumor-bearing NSG mice with activated NK cells led to significant reductions in both intratumoral CSCs and tumor burden. Taken together, these data from multiple preclinical models, including a strong reliance on primary human cancer specimens, provide compelling preclinical evidence that activated NK cells preferentially target cancer cells with a CSC phenotype, highlighting the translational potential of NK immunotherapy as part of a combined modality approach for refractory solid malignancies.


Yolk-shell Fe(0)@SiO2 nanoparticles as nanoreactors for fenton-like catalytic reaction.
Yolk-shell nanoparticles (YSNs) with active metal cores have shown promising applications in nanoreactors with excellent catalytic performance. In this work, Fe(0)@SiO2 YSNs were synthesized by a sequential "two-solvents" impregnation-reduction approach. Specifically, FeSO4 aqueous solution was introduced into the preformed hollow mesoporous silica spheres (HMSS), dispersed in n-hexane, via a "two-solvent" impregnation way. Subsequently, aqueous solution of sodium borohydride (NaBH4) was introduced into the cavity of HMSS by the same way, leading to the formation of Fe core inside the HMSS through the reaction between Fe(2+) and NaBH4. The resulting Fe(0)@SiO2 YSNs possess distinctive structures, including active cores, accessible mesoporous channels, protective shells, and hollow cavities. To present the catalytic performance of YSNs nanoreactors, Fenton-like catalytic oxidation of phenol was chosen as the model catalysis reaction. In addition to the Fe(0)@SiO2 YSNs, two other materials were also applied to the catalytic system for comparison, including Fe@SiO2 composites with iron nanoparticles sticking on the outer shells of HMSS (Fe@SiO2-DI) and bare iron nanoparticles without HMSS (bare Fe(0)), respectively. The catalytic results show that Fe(0)@SiO2 YSNs exhibit higher catalytic rate toward phenol removal at 2-fold and 4-fold as compared to that of Fe@SiO2-DI and bare Fe(0), indicating the outstanding catalytic property of YSNs nanoreactors. To further clarify the relationship between catalytic properties and structural characteristics, the adsorption experiments of the three samples were also performed in the absence of H2O2. Other than catalytic results, Fe(0)@SiO2 YSNs show slightly higher adsorption efficiency than the other two samples, indicating the accessibility of nanoreactors. This result demonstrates that the removal of phenol in the oxidation system of Fe(0)@SiO2 YSNs may have contributed to the structure-enhanced effect of YSNs as nanoreactors.


Public perception of geothermal power plants in Korea following the Pohang earthquake: A social representation theory study.
This study sought to determine how the residents of Pohang, Korea, perceive geothermal plants after the 2017 Pohang earthquake by applying social representation theory through a mixed-method approach incorporating qualitative and quantitative research. The residents' perception of the geothermal plant was largely anchored to their perception of nuclear power plants. At the time of the Gyeongju earthquake in 2016, public discourse on nuclear accidents developed and was thereafter perpetuated by the Pohang earthquake victims via cognitive anchoring. The survey results demonstrated that Pohang residents had a significantly negative opinion on geothermal plants regardless of safety, climate change mitigation, and economic factors. Upon analyzing the respondents' energy preferences through factor analysis, geothermal power plants were found to aggregate in the same category as nuclear power plants. This result statistically confirms that Pohang residents associate geothermal power plants with the risk discourse on nuclear power plants.


Mannitol or hypertonic saline in the setting of traumatic brain injury: What have we learned?
Intracranial hypertension, defined as an intracranial pressure (ICP) >20 mmHg for a period of more than 5 min, worsens neurologic outcome in traumatic brain injury (TBI). While several mechanisms contribute to poor outcome, impaired cerebral perfusion appears to be a highly significant common denominator. Management guidelines from the Brain Trauma Foundation recommend measuring ICP to guide therapy. In particular, hyperosmolar therapy, which includes mannitol or hypertonic saline (HTS), is frequently administered to reduce ICP. Currently, mannitol (20%) is considered the gold standard hyperosmolar agent. However, HTS is increasingly used in this setting. This review sought to compare the efficacy of mannitol to HTS in severe TBI. The PubMed database was used to systematically search for articles comparing mannitol to HTS in severe TBI. The following medical subject headings were used: HTS, sodium lactate, mannitol, ICP, intracranial hypertension, and TBI. We included both prospective and retrospective randomized controlled studies of adult patients with intracranial hypertension as a result of severe TBI who received hyperosmolar therapy. Out of 45 articles, seven articles were included in our review: 5 were prospective, randomized trials; one was a prospective, nonrandomized trial; and one was a retrospective, cohort study. While all seven studies found that both mannitol and HTS were effective in reducing ICP, there was heterogeneity with regard to which agent was most efficacious.