Incidences of Severe Hypoglycemia and Diabetic Ketoacidosis and Prevalence of Microvascular Complications Stratified by Age and Glycemic Control in U.S. Adult Patients With Type 1 Diabetes: A Real-World Study.
To assess the burden of disease for adults with type 1 diabetes in a U.S. electronic health record database by evaluating acute and microvascular complications stratified by age and glycemic control. This is a retrospective observational study of adults with type 1 diabetes (1 July 2014-30 June 2016) classified using a validated algorithm, with disease duration ≥24 months and, during a 12-month baseline period, not pregnant and having one or more insulin prescriptions and one or more HbA1c measurements. Demographic characteristics, acute complications (severe hypoglycemia [SH], diabetic ketoacidosis [DKA]), and microvascular complications (neuropathy, nephropathy, retinopathy) were stratified by age (18-25, 26-49, 50-64, ≥65 years) and glycemic control (HbA1c <7%, 7% to <9%, ≥9%). Of 31,430 patients, ∼20% had HbA1c <7%. Older patients had lower HbA1c values than younger patients (P < 0.001). Patients with poor glycemic control had the highest annual incidence of SH (4.2%, 4.0%, and 8.3%) and DKA (1.3%, 2.8%, and 15.8%) for HbA1c <7%, 7% to <9%, and ≥9% cohorts, respectively (both P < 0.001), and a higher prevalence of neuropathy and nephropathy (both P < 0.001). For adults with type 1 diabetes, glycemic control appears worse than previously estimated. Rates of all complications increased with increasing HbA1c. Compared with HbA1c <7%, HbA1c ≥9% was associated with twofold and 12-fold higher incidences of SH and DKA, respectively. Younger adults had more pronounced higher risks of SH and DKA associated with poor glycemic control than older adults.


Gender differences in disordered eating and weight dissatisfaction in Swiss adults: which factors matter?
Research results from large, national population-based studies investigating gender differences in weight dissatisfaction and disordered eating across the adult life span are still limited. Gender is a significant factor in relation to weight dissatisfaction and disordered eating. However, the reasons for gender differences in these conditions are still poorly understood. The aim of this study was to examine gender differences in weight dissatisfaction and disordered eating in the general Swiss adult population and to identify gender-specific risk factors. The study population consisted of 18156 Swiss adults who completed the population-based Swiss Health Survey 2007. Self-reported weight dissatisfaction, disordered eating and associated risk factors were assessed. In order to examine whether determinants of weight dissatisfaction and disordered eating (dieting to lose weight, binge eating, and irregular eating) differ in men and women, multivariate logistic regressions were applied separately for women and men. Although more men than women were overweight, more women than men reported weight dissatisfaction. Weight category, smoking status, education, and physical activity were significantly associated with weight dissatisfaction in men and women. In women, nationality and age were also significant factors. Gender-specific risk factors such as physical activity or weight category were identified for specific disordered eating behaviours. The results suggest that gender specific associations between predictors and disordered eating behaviour should be considered in the development of effective prevention programs against disordered eating.


[Binding of tylosin, tilmicosin and oxytetracycline to proteins from honeybees, larvae and beehive products].
American Foulbrood (AFB) caused by the spore-forming bacterium Paenibacillus larvae is the most serious disease of bacterial origin affecting larvae and pupae of honeybees. Antibiotics are used in many countries for the control of AFB in high incidence areas, but their misuse may lead to antibiotic resistance of bacterial strains and honey contamination. The objective of the present work was to determine, through a biological method, the protein binding of tylosin, tilmicosin and oxytetracycline to worker jelly; honey; pollen; adult bees and larvae in order to propose their kinetic routes. The sensitivity limit of the technique used was 0.05 μg/ml for tylosin and tilmicosin and 0.01 μg/ml for oxytetracycline, respectively. The method had intra and inter-assay correlation coefficients over 0.90, respectively and a coefficient variation of intra-and inter-assay for all antibiotics and processed samples under 5%. Tylosin and oxytetracycline presented lower percentages of protein binding in tissues and hive products (average 15%) in relation to those observed for tilmicosin (29%). In conclusion, tylosin is useful for AFB control in honey bee colonies due to its chemical characteristics, antimicrobial activity and levels of protein binding in bees, larvae, and beehive products.


Use of cetirizine to investigate non-H1 effects of second-generation antihistamines.
In addition to their increased potency as H1 blockers and their nonsedating effects, the second-generation antihistamines have other unusual and potentially beneficial properties. Evidence is accumulating from several laboratories that at least one of these agents under investigation, cetirizine, may be effective in inhibiting the late reaction. The Johns Hopkins group showed that during the cutaneous late phase response (LPR), histamine release was not altered by cetirizine, 20 mg, pretreatment. The most dramatic effect of cetirizine was attenuation of inflammatory cell migration into the chamber. Eosinophils, neutrophils, and basophils were reduced by about 75% during hours 6 to 8. It can be concluded that cetirizine influences the LPR by causing a reduction in the inflammatory cell infiltrate. Cetirizine, 10 mg, orally once a day also induced a significant decrease in the wheal and flare skin reactions caused by pollen, histamine, and compound 48/80. Cetirizine inhibited eosinophil recruitment and platelet-activating factor (PAF) in skin chambers 24 hours after pollen challenge. We and others have studied the mechanisms of this effect. The release of eosinophil peroxidase induced by PAF and formyl-methionyleucyl/phenylalanine was not attenuated by cetirizine. At therapeutic concentrations, however, cetirizine has a potent inhibitory action in vitro on eosinophil chemotaxis induced either by formyl-methionyleucyl/phenylalanine or PAF and also on IgE-dependent stimulation of platelets. In a separate study in patients with chronic urticaria, cetirizine markedly reduced both the immediate wheal and flare induced by PAF and the delayed reaction at six hours. These results suggest that cetirizine acts on eosinophil migration to inhibit the late reaction.


Endoscopic Removal of an Epithelial Cyst of the Zygoma.
This study elaborates a brief overview of epithelial cysts in the bones of the skull and describes an unusual patient with an epidermoid cyst of the zygoma. This report focuses on an endoscopic preauricular infratemporal fossa approach for resection of a left epidermoid cyst of the zygoma. Preoperative magnetic resonance imaging and computed tomography imaging as well as intraoperative endoscopic images and movie were demonstrated. The epithelial cyst was successfully removed from the patient using a minimally invasive approach utilizing endoscopes. No complications were encountered. The possible causes of this rare presentation and minimal surgical removal utilizing endoscopes were discussed in this study. According to the authors, this is the first reported study of endoscopic removal of an epidermoid cyst from the zygomatic root. The patient's previous surgical history of a tympanoplasty could have contributed to the unusual location of the lesion.


Enzymatic incorporation and utilization of an emissive 6-azauridine.
To display favorable fluorescent properties, the non-emissive native nucleosides need to be modified. Here we present a motif that relies on conjugating 5-membered aromatic heterocycles (e.g., thiophene) to a 6-azapyrimidine (1,2,4-triazine) core. Synthetic accessibility and desirable photophysical properties make these nucleosides attractive candidates for enzymatic incorporation and biochemical assays. While 6-azauridine triphosphate is known to be poorly tolerated by polymerases in RNA synthesis, we illustrate that conjugating a thiophene ring at position 5 overcomes such limitations, facilitating its T7 RNA polymerase-mediated in vitro transcription incorporation into RNA constructs. We further show that the modified transcripts can be ligated to longer oligonucleotides to form singly modified RNAs, as illustrated for an A-site hairpin model RNA construct, which was employed to visualize aminoglycoside antibiotics binding.


Partner experiences of breast reconstruction post mastectomy.
The aim of this small-scale, qualitative study was to explore partners' experiences of breast reconstruction after mastectomy. Interviews with 6 men, analysed using thematic analysis identified key themes of Involvement in the Decision-Making Process, Giving and Seeking Support, Opportunity to Talk, and Adapting Behaviour. Whilst most participants wanted to be involved in the decision-making process, they reported having experienced barriers to this, including difficulty in talking to others and expressing their true feelings about the surgery. Participants described difficulties with intimate contact after their partner had undergone surgery due to reactions to the appearance and feel of the reconstructed breast and a fear of hurting their partner and of reopening scars. This study highlights the need to recognise the role of the partner's involvement and of their own experiences during this time. The findings are of value to healthcare professionals, in particular specialist nurses, who work with women undergoing breast reconstruction and indicate areas where further research is needed.


Gender differences in the trajectories of late-life depressive symptomology and probable depression in the years prior to death.
Gender differences in depression are well established. Whether these differences persist into late life and in the years preceding death is less clear. There is a suggestion that there is no increased likelihood of depression in late life, but that there is an increase in depressive symptomology, particularly with proximity to death. We compared trajectories of probable depression and depressive symptomology between men and women over age and distance-to-death metrics to determine whether reports of depressive symptoms are more strongly related to age or mortality. Participants (N = 2,852) from the Dynamic Analyses to Optimise Ageing (DYNOPTA) project had a mean age of 75 years (SD = 5.68 years) at baseline and were observed for up to 16 years prior to death. Multi-level regression models estimated change in depressive symptomology and probable depression over two time metrics, increasing age, and distance-to-death. Increases in depressive symptomology were reported over increasing age and in the years approaching death. Only male participants reported increased probable depression in the years preceding death. Models that utilized distance-to-death metrics better represented changes in late-life depression, although any changes in depression appear to be accounted for by co-varying physical health status. As death approaches, there are increases in the levels of depressive symptomology even after controlling for socio-demographic and health covariates. In line with increases in suicide rates in late life, male participants were at greater risk of reporting increases in depressive symptomology.


Phenological patterns of flowering across biogeographical regions of Europe.
Long-term changes of plant phenological phases determined by complex interactions of environmental factors are in the focus of recent climate impact research. There is a lack of studies on the comparison of biogeographical regions in Europe in terms of plant responses to climate. We examined the flowering phenology of plant species to identify the spatio-temporal patterns in their responses to environmental variables over the period 1970-2010. Data were collected from 12 countries along a 3000-km-long, North-South transect from northern to eastern Central Europe.Biogeographical regions of Europe were covered from Finland to Macedonia. Robust statistical methods were used to determine the most influential factors driving the changes of the beginning of flowering dates. Significant species-specific advancements in plant flowering onsets within the Continental (3 to 8.3 days), Alpine (2 to 3.8 days) and by highest magnitude in the Boreal biogeographical regions (2.2 to 9.6 days per decades) were found, while less pronounced responses were detected in the Pannonian and Mediterranean regions. While most of the other studies only use mean temperature in the models, we show that also the distribution of minimum and maximum temperatures are reasonable to consider as explanatory variable. Not just local (e.g. temperature) but large scale (e.g. North Atlantic Oscillation) climate factors, as well as altitude and latitude play significant role in the timing of flowering across biogeographical regions of Europe. Our analysis gave evidences that species show a delay in the timing of flowering with an increase in latitude (between the geographical coordinates of 40.9 and 67.9), and an advance with changing climate. The woody species (black locust and small-leaved lime) showed stronger advancements in their timing of flowering than the herbaceous species (dandelion, lily of the valley). In later decades (1991-2010), more pronounced phenological change was detected than during the earlier years (1970-1990), which indicates the increased influence of human induced higher spring temperatures in the late twentieth century.


Can I join in? Multiple case study investigation of play performance generalisation for children with autism spectrum disorder from dyad to triad.
Children with autism spectrum disorder (ASD) have difficulties with play, social interaction with peers and generalisation of intervention outcomes. The Ultimate Guide to Play, Language and Friendship (PLF) has demonstrated effectiveness in improving play performance of children with ASD and their typically developing (TD) peers. The aim of this investigation was to examine the changes in play performance when an additional TD child is added to an existing dyad of a child with ASD and a TD playmate to inform future delivery and adaptations of the intervention. Participants in this multiple case study design were five children with ASD and their TD peer who completed a dyad intervention as part of a randomised control trial investigation of the PLF and an additional TD peer who joined the play dyad. A trained occupational therapist delivered an adapted version of the PLF to the triad over four clinic sessions. An independent rater scored each child (N = 15) on The Test of Playfulness at pre- and post-triad intervention. Line graphs were used to examine case data and compare to dyad play performance and patterns of interaction. Four of the five children with ASD generalised their play performance from the dyad to the triad social environment. However, the triad intervention did not demonstrate improvements in play performance. The play performance scores for the children with ASD and their TD peers were variable and demonstrated changes in their play pattern from the dyad to the triad. This investigation delivered preliminary evidence of play performance generalisation from a dyad to a triad with TD peers for children with ASD. Careful consideration of characteristics of all playmates is recommended for delivering the intervention to support play performance of children with ASD.


Variants of C-C motif chemokine 22 (CCL22) are associated with susceptibility to atopic dermatitis: case-control studies.
Atopic dermatitis (AD) is a common inflammatory skin disease caused by multiple genetic and environmental factors. AD is characterized by the local infiltration of T helper type 2 (Th2) cells. Recent clinical studies have shown important roles of the Th2 chemokines, CCL22 and CCL17 in the pathogenesis of AD. To investigate whether polymorphisms of the CCL22 gene affect the susceptibility to AD, we conducted association studies and functional studies of the related variants. We first resequenced the CCL22 gene and found a total of 39 SNPs. We selected seven tag SNPs in the CCL22 gene, and conducted association studies using two independent Japanese populations (1(st) population, 916 cases and 1,032 controls; 2(nd) population 1,034 cases and 1,004 controls). After the association results were combined by inverse variance method, we observed a significant association at rs4359426 (meta-analysis, combined P = 9.6×10⁻⁶; OR, 0.74; 95% CI, 0.65-0.85). Functional analysis revealed that the risk allele of rs4359426 contributed to higher expression levels of CCL22 mRNA. We further examined the allelic differences in the binding of nuclear proteins by electrophoretic mobility shift assay. The signal intensity of the DNA-protein complex derived from the G allele of rs223821, which was in absolute LD with rs4359426, was higher than that from the A allele. Although further functional analyses are needed, it is likely that related variants play a role in susceptibility to AD in a gain-of-function manner. Our findings provide a new insight into the etiology and pathogenesis of AD.


Physicians and the pharmaceutical industry: is a gift ever just a gift?
Controversy exists over the fact that physicians have regular contact with the pharmaceutical industry and its sales representatives, who spend a large sum of money each year promoting to them by way of gifts, free meals, travel subsidies, sponsored teachings, and symposia. To identify the extent of and attitudes toward the relationship between physicians and the pharmaceutical industry and its representatives and its impact on the knowledge, attitudes, and behavior of physicians. A MEDLINE search was conducted for English-language articles published from 1994 to present, with review of reference lists from retrieved articles; in addition, an Internet database was searched and 5 key informants were interviewed. A total of 538 studies that provided data on any of the study questions were targeted for retrieval, 29 of which were included in the analysis. Data were extracted by 1 author. Articles using an analytic design were considered to be of higher methodological quality. Physician interactions with pharmaceutical representatives were generally endorsed, began in medical school, and continued at a rate of about 4 times per month. Meetings with pharmaceutical representatives were associated with requests by physicians for adding the drugs to the hospital formulary and changes in prescribing practice. Drug company-sponsored continuing medical education (CME) preferentially highlighted the sponsor's drug(s) compared with other CME programs. Attending sponsored CME events and accepting funding for travel or lodging for educational symposia were associated with increased prescription rates of the sponsor's medication. Attending presentations given by pharmaceutical representative speakers was also associated with nonrational prescribing. The present extent of physician-industry interactions appears to affect prescribing and professional behavior and should be further addressed at the level of policy and education.


Inhibition of 5 alpha-reductase activity in human skin by zinc and azelaic acid.
The effects of zinc sulphate and azelaic acid on 5 alpha-reductase activity in human skin were studied using an in vitro assay with 1,2[3H]-testosterone as substrate. When added at concentrations of 3 or 9 mmol/l, zinc was a potent inhibitor of 5 alpha-reductase activity. At high concentrations, zinc could completely inhibit the enzyme activity. Azelaic acid was also a potent inhibitor of 5 alpha-reductase; inhibition was detectable at concentrations as low as 0.2 mmol/l and was complete at 3 mmol/l. An additive effect of the two inhibitors was observed. Vitamin B6 potentiated the inhibitory effect of zinc, but not of azelaic acid, suggesting that two different mechanisms are involved. When the three substances were added together at very low concentrations which had been shown to be ineffective alone, 90% inhibition of 5 alpha-reductase activity was obtained. If this inhibition is confirmed in vivo, zinc sulphate combined with azelaic acid could be an effective agent in the treatment of androgen related pathology of human skin.


Evidence that microRNA precursors, unlike other non-coding RNAs, have lower folding free energies than random sequences.
Most non-coding RNAs are characterized by a specific secondary and tertiary structure that determines their function. Here, we investigate the folding energy of the secondary structure of non-coding RNA sequences, such as microRNA precursors, transfer RNAs and ribosomal RNAs in several eukaryotic taxa. Statistical biases are assessed by a randomization test, in which the predicted minimum free energy of folding is compared with values obtained for structures inferred from randomly shuffling the original sequences. In contrast with transfer RNAs and ribosomal RNAs, the majority of the microRNA sequences clearly exhibit a folding free energy that is considerably lower than that for shuffled sequences, indicating a high tendency in the sequence towards a stable secondary structure. A possible usage of this statistical test in the framework of the detection of genuine miRNA sequences is discussed.


The effect of marker choice on estimated levels of introgression across an avian (Pipridae: Manacus) hybrid zone.
Hybrid zones are often characterized by narrow, coincident clines for diverse traits, suggesting that little introgression occurs across them. However, this pattern may result from a bias in focussing on traits that are diagnostic of parental populations. Such choice of highly differentiated traits may cause us to overlook differential introgression in nondiagnostic traits and to distort our perception of hybrid zones. We tested this hypothesis in an avian hybrid zone by comparing cline structure in two sets of molecular markers: isozyme and restriction fragment length polymorphism markers chosen for differentiation between parental forms, and microsatellite markers chosen for polymorphism. Two cline-fitting methods showed that cline centre positions of microsatellite alleles were more variable than those of isozyme and restriction fragment length polymorphism markers, and several were significantly shifted from those of the diagnostic markers. Cline widths of microsatellite alleles were also variable and two- to eightfold wider than those of the diagnostic markers. These patterns are consistent with the idea that markers chosen for differentiation are more likely to be under purifying selection, and studies focussed on these markers will underestimate overall introgression across hybrid zones. Our results suggest that neutral and positively selected alleles may introgress freely across many hybrid zones without altering perceived boundaries between hybridizing forms.


Maternal environment and craniofacial growth: geometric morphometric analysis of mandibular shape changes with in utero thyroxine overexposure in mice.
An estimated 3% of US pregnancies are affected by maternal thyroid dysfunction, with between one and three of every 1000 pregnancies being complicated by overactive maternal thyroid levels. Excess thyroid hormones are linked to neurological impairment and excessive craniofacial variation, affecting both endochondral and intramembranous bone. Using a geometric morphometric approach, this study evaluates the role of in utero thyroxine overexposure on the growth of offspring mandibles in a sample of 241 mice. Canonical variate analysis utilized 16 unilateral mandibular landmarks obtained from 3D micro-computed tomography to assess shape changes between unexposed controls (n = 63) and exposed mice (n = 178). By evaluating shape changes in the mandible among three age groups (15, 20 and 25 days postnatal) and different dosage levels (low, medium and high), this study found that excess maternal thyroxine alters offspring mandibular shape in both age- and dosage-dependent manners. Group differences in overall shape were significant (P < 0.001), and showed major changes in regions of the mandible associated with muscle attachment (coronoid process, gonial angle) and regions of growth largely governed by articulation with the cranial base (condyle) and occlusion (alveolus). These results compliment recent studies demonstrating that maternal thyroxine levels can alter the cranial base and cranial vault of offspring, contributing to a better understanding of both normal and abnormal mandibular development, as well as the medical implications of craniofacial growth and development.


Inhibition of lymphocyte proliferation by detergent-solubilized mouse liver membranes.
The role of phenomena analogous to fibroblast contact inhibition in lymphocyte growth regulation is controversial, although it is clear that direct cell-cell contact is vital to immunoregulation and accessory cell function. An extract of mouse liver plasma membrane proteins, referred to as suppressive liver extract (SLE), that suppresses the growth of 3T3 fibroblasts also inhibited the mitogen-induced proliferation of murine lymphocytes. A dose of 20 micrograms/ml SLE was less than 95% suppressive of proliferation in both mouse T and mouse B cells treated with a variety of mitogens. B cell growth factor, while increasing DNA synthesis overall in mitogen-stimulated B cells, did not change the extent of SLE suppression, which suggests that the SLE does not interfere with lymphocyte-growth factor interactions. In exploring a sequence of B cell activation events, we discovered that SLE had no effect on the early activation event of increased phosphatidylinositol turnover. Blastogenesis, however, was inhibited in mitogen-stimulated, SLE-treated B cells. The maximum suppressive effect was observed if the SLE was added within 8-12 h of the mitogenic stimulus. SLE did not affect the viability of cells in culture. These results point to a possible unity of regulatory mechanisms between contact inhibition in fibroblasts and the processes of immunoregulation.


Baseline Evaluation of a Participatory Mobile Health Intervention for Dengue Prevention in Sri Lanka.
Challenges posed by infectious disease outbreaks have led to a range of participatory mobile phone-based innovations that use the power of crowdsourcing for disease surveillance. However, the dynamics of participatory behavior by crowds in such interventions have yet to be examined. This article reports results from a baseline evaluation of one such intervention called Mo-Buzz, a mobile-based crowdsource-driven socially mediated system developed to address gaps in dengue surveillance and education in Colombo, Sri Lanka. We conducted a 30-minute cross-sectional field survey (N = 404) among potential users of Mo-Buzz in Colombo. We examined individual, institutional, and cultural factors that influence their potential intention-to-use Mo-Buzz and assessed if these factors varied by demographic factors. Descriptive analysis revealed high perceived ease-of-use (PEOU; M = 3.81, SD = 0.44), perceived usefulness (PU; M = 4.01, SD = 0.48), and intention-to-use (PI; M = 3.91, SD = 0.46) among participants. Analysis of variance suggested participants in the 31 to 40 years age group reported highest PEOU, whereas the oldest group reported high perceived institutional efficacy (M = 3.59, SD = 0.64) and collectivistic tendencies. Significant differences (at the p < .05 level) were also found by education and income. Regression analysis demonstrated that PU, behavioral control, institutional efficacy, and collectivism were significant predictors of PI. We concluded that despite high overall PI, future adoption and use of Mo-Buzz will be shaped by a complex mix of factors at different levels of the public health ecology. Implications of study findings from theoretical and practical perspectives related to the future adoption of mobile-based participatory systems in public health are discussed and ideas for a future research agenda presented.


Utilization, spending, and price trends for short- and long-acting Beta-agonists and inhaled corticosteroids in the medicaid program, 1991-2010.
Asthma is a chronic respiratory disease that afflicts millions of people and accounts for substantial utilization of healthcare resources in most industrialized countries, including in the United States. However, the exact cost and utilization of anti-asthma medications in Medicaid in the past 2 decades have not been well studied. Considering the safety issues surrounding the long-acting beta-agonists, guideline updates, and the increase in asthma prevalence, understanding anti-asthma medication prescribing trends is important to payers and patients. The purpose of this study was to analyze the utilization and spending trends for anti-asthmatic agents in the US Medicaid program over the past 2 decades. This study was based on a retrospective, descriptive analysis of trends in utilization of and spending on anti-asthma medications, including short-acting beta-agonists, inhaled corticosteroids, long-acting beta-agonists, and inhaled corticosteroid/long-acting beta-agonist combinations. Quarterly utilization and expenditure data were obtained from the national Medicaid pharmacy files provided by the Centers for Medicare & Medicaid Services from quarter 1 of 1991 through quarter 2 of 2010. Average reimbursement per prescription was calculated each quarter as a proxy for drug price. The total number of prescriptions for the studied anti-asthma medications rose from 8.9 million in 1991 to 15.6 million in 2009, peaking at 20.8 million in 2005, the year before Medicare and Medicaid dual-eligible beneficiaries were moved to Medicare Part D. From 1991 to 2009, Medicaid spending on anti-asthma medications overall rose from $180.7 million to $1.3 billion, and spending on inhaled corticosteroid/long-acting beta-agonist combinations rose from $52.8 million in 2001-their first year on the market-to $411.7 million in 2009. The average price per prescription has risen in all the anti-asthma drug classes: overall, spending per prescription has increased 4-fold between 1991 and 2009, significantly faster than the consumer price index (57.5%) over the same period. In quarter 2 of 2010, Medicaid spent more on the combination medication fluticasone-salmeterol-$60 million-than on any other anti-asthma medication. Anti-asthma medications are a major and growing expense for state Medicaid programs and can be expected to be the same for Medicare Part D in the future. Increased disease prevalence has in part contributed to the rise in pharmacotherapy cost. Nevertheless, drug therapy is crucial for managing asthma and asthma exacerbations.


A novel gene-wide haplotype at the macrophage migration inhibitory factor (MIF) locus is associated with endometrioma.
Endometriosis is a common complex gynecological disorder that may result in infertility. Macrophage migration inhibitory factor (MIF) is a key pro-inflammatory cytokine that is overexpressed in endometriosis tissues. However, hitherto, no study tested the possible relevancy at genetic level. The aim of this study was to evaluate MIF polymorphisms and possible associations between haplotype of the gene and endometrioma. In this experiment, 115 patients with confirmed endometrioma and 120 of women who were not diagnosed with endometrioma were recruited for this case-control genetic association study. The coding region of MIF was resequenced to detect variations of potential significance. Restriction fragment length polymorphism was used to type the -173 G/C (rs755622) promoter Single nucleotide polymorphism (SNP). Haplotype analyses were then undertaken to assess the effect of genetic variations. We detected one functional SNP in promoter (rs755622) and non-functional mutations across the gene including (rs2096525, rs182012324, rs33958703 and rs2070766) in our samples. However, haplotype analysis showed a significant association between MIF and endometrioma where a single haplotype CC carrying only the minor allele at -173 G/C was significantly over-represented in the patients group (P = 0.007) and remained significant even after correction for (Bonferroni adjusted P = 0.028). We report a strong linkage between a novel MIF haplotype and endometrioma. This association is consistent with expression data at both transcript and protein levels suggesting the -173C/G promoter as a critical factor.


Prevalence and factors associated with 2009 to 2011 influenza vaccinations at a university medical center.
Information on the rates and factors associated with influenza vaccinations, although limited, is important because it can inform the development of effective vaccination campaigns in a university medical center setting. A study was conducted in 2011 to identify individual and organizational level barriers and facilitators to influenza vaccination among clinical and nonclinical personnel (N = 428) from a major university medical center. Seventy-one percent of clinical personnel (n = 170) reported pandemic H1N1 vaccination compared with 27% of nonclinical personnel (n = 258), even though vaccine was made widely available to all personnel at no cost. Similarly, disparate rates between clinical and nonclinical personnel were noted for the 2009/2010 seasonal influenza vaccine (82% vs 42%, respectively) and 2010/2011 combination (pandemic plus seasonal) influenza vaccine (73% vs 28%, respectively). Factors associated with pandemic vaccination in nonclinical personnel included the following: high level of influenza-related knowledge, concern regarding influenza contagion, history of previous influenza vaccinations or influenza illness, participation in vaccine-related training, and awareness of the institution's written pandemic plan. For clinicians, past history of seasonal influenza vaccination was associated with pandemic vaccination. For all participants, taking any 1 or more of the 3 influenza vaccines available in 2009 to 2011 was associated with intent to take a hypothetical future novel pandemic vaccine (odds ratio, 6.7; 95% confidence interval: 4.32-10.44; P < .001). Most of the risk factors associated with lack of vaccination uptake are amenable to organizational strategies.


Intended versus inferred care after PET performed for initial staging in the National Oncologic PET Registry.
The National Oncologic PET Registry (NOPR) collected data on intended management before and after PET in cancer patients. We have previously reported that PET was associated with a change in intended management of about one third of patients and was consistent across cancer types. It is uncertain if intended management plans reflect the actual care these patients received. One approach to assess actual care received is using administrative claims to categorize the type and timing of clinical services. NOPR data from 2006 to 2008 were linked to Medicare claims for consenting patients aged 65 y or older undergoing initial-staging PET scanning for bladder, ovarian, pancreatic, small cell lung, or stomach cancers. We determined the 60-d agreement between claims-inferred care and NOPR treatment plans. Patients (n = 4,661) were assessed, and 30%-52% had metastatic disease. Planned treatments were about two-thirds monotherapy, of which 46% was systemic therapy only, and one-third combinations. Claims paid by 60 d confirmed the NOPR plan of any systemic therapy, radiotherapy, or surgery in 79.3%, 64.7%, and 63.6%, respectively. Single-mode plans were much more often confirmed: systemic therapy in more than 85% of patients with ovarian, pancreatic, and small cell lung cancers and surgery in more than 73% of those with bladder, pancreatic, and stomach cancers. Intended combination treatments had claims for both in only 28% of patients receiving surgery-based combinations and in 55% receiving chemoradiotherapy. About 90% of patients with NOPR-planned systemic therapy had evaluation or management claims from a medical oncologist. An age of less than 75 y was associated more often with confirmation of chemotherapy, less often for radiotherapy but not with confirmation of surgery. Performance status or comorbidity did not explain confirmation rates within action categories, but confirmation rates were higher if the referrer specialized in the planned treatment. Claims confirmations of NOPR intent for initial staging were widely variable but were higher than previously reported for restaging PET, suggesting that measuring change in intended management is a reasonable method for assessing the impact diagnostic tests have on actual care.


Time-efficient intervention to improve older adolescents' cardiorespiratory fitness: findings from the 'Burn 2 Learn' cluster randomised controlled trial.
Cardiorespiratory fitness (CRF) is an important marker of current and future health status. The primary aim of our study was to evaluate the impact of a time-efficient school-based intervention on older adolescents' CRF. Two-arm cluster randomised controlled trial conducted in two cohorts (February 2018 to February 2019 and February 2019 to February 2020) in New South Wales, Australia. Participants (N=670, 44.6% women, 16.0±0.43 years) from 20 secondary schools: 10 schools (337 participants) were randomised to the Burn 2 Learn (B2L) intervention and 10 schools (333 participants) to the control. Teachers in schools allocated to the B2L intervention were provided with training, resources, and support to facilitate the delivery of high-intensity interval training (HIIT) activity breaks during curriculum time. Teachers and students in the control group continued their usual practice. The primary outcome was CRF (20 m multi-stage fitness test). Secondary outcomes were muscular fitness, physical activity, hair cortisol concentrations, mental health and cognitive function. Outcomes were assessed at baseline, 6 months (primary end-point) and 12 months. Effects were estimated using mixed models accounting for clustering. We observed a group-by-time effect for CRF (difference=4.1 laps, 95% CI 1.8 to 6.4) at the primary end-point (6 months), but not at 12 months. At 6 months, group-by-time effects were found for muscular fitness, steps during school hours and cortisol. Implementing HIIT during curricular time improved adolescents' CRF and several secondary outcomes. Our findings suggest B2L is unlikely to be an effective approach unless teachers embed sessions within the school day. Australian New Zealand Clinical Trials Registry (ACTRN12618000293268).


Waterborne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt.
At least 325 water-associated outbreaks of parasitic protozoan disease have been reported. North American and European outbreaks accounted for 93% of all reports and nearly two-thirds of outbreaks occurred in North America. Over 30% of all outbreaks were documented from Europe, with the UK accounting for 24% of outbreaks, worldwide. Giardia duodenalis and Cryptosporidium parvum account for the majority of outbreaks (132; 40.6% and 165; 50.8%, respectively), Entamoeba histolytica and Cyclospora cayetanensis have been the aetiological agents in nine (2.8%) and six (1.8%) outbreaks, respectively, while Toxoplasma gondii and Isospora belli have been responsible for three outbreaks each (0.9%) and Blastocystis hominis for two outbreaks (0.6%). Balantidium coli, the microsporidia, Acanthamoeba and Naegleria fowleri were responsible for one outbreak, each (0.3%). Their presence in aquatic ecosystems makes it imperative to develop prevention strategies for water and food safety. Human incidence and prevalence-based studies provide baseline data against which risk factors associated with waterborne and foodborne transmission can be identified. Standardized methods are required to maximize public health surveillance, while reporting lessons learned from outbreaks will provide better insight into the public health impact of waterborne pathogenic protozoa.


The expression of minichromosome maintenance protein-2 in normal and abnormal megakaryocytes and comparison with the proliferative marker Ki-67.
The minichromosome maintenance (Mcm) and Cdc6 proteins are important regulators of eucaryotic DNA replication. In most normal tissues, a similar proportion of cells express Mcm-2 and Ki-67. The present study showed that in both normal and abnormal states, the proportion of megakaryocytes expressing Mcm-2 is roughly seven times as many as those that express Ki-67. This is likely to be related to the process of endomitosis and endoreduplication. We also demonstrated that a significantly lower proportion of megakaryocytes in myelodysplastic syndrome express Mcm-2. These findings provide new insights into megakaryocyte biology.


Curved TiO2 Nanoparticles in Water: Short (Chemical) and Long (Physical) Range Interfacial Effects.
In most technological applications, nanoparticles are immersed in a liquid environment. Understanding nanoparticles/liquid interfacial effects is extremely relevant. This work provides a clear and detailed picture of the type of chemistry and physics taking place at the prototypical TiO2 nanoparticles/water interface, which is crucial in photocatalysis and photoelectrochemistry. We present a multistep and multiscale investigation based on hybrid density functional theory (DFT), density functional tight-binding, and quantum mechanics/molecular mechanics calculations. We consider increasing water partial pressure conditions from ultra-high vacuum up to the bulk water environment. We first investigate single water molecule adsorption modes on various types of undercoordinated sites present on a realistic curved nanoparticle (2-3 nm) and then, by decorating all the adsorption sites, we study a full water monolayer to identify the degree of water dissociation, the Brønsted-Lowry basicity/acidity of the nanoparticle in water, the interface effect on crystallinity, surface energy, and electronic properties, such as the band gap and work function. Furthermore, we increase the water coverage by adding water multilayers up to a thickness of 1 nm and perform molecular dynamics simulations, which evidence layer structuring and molecular orientation around the curved nanoparticle. Finally, we clarify whether these effects arise as a consequence of the tension at the water drop surface around the nanosphere by simulating a bulk water up to a distance of 3 nm from the oxide surface. We prove that the nanoparticle/water interfacial effects go rather long range since the dipole orientation of water molecules is observed up to a distance of 5 Å, whereas water structuring extends at least up to a distance of 8 Å from the surface.


Genetic differentiation between two varieties of Oreocharis benthamii (Gesneriaceae) in sympatric and allopatric regions.
The pattern of genetic differentiation between diverging species receives much attention as one of the key observable features of speciation. It has often been suggested that introgression between closely related species occurs commonly where their distributions overlap, leading to their becoming more morphologically and genetically similar, but there are a few opposite results. However, most of these studies have been carried out with animals and separate species; few have looked at intraspecific cases, especially in plants. Here, we conduct a comparative study on patterns of genetic differentiation among populations of two varieties of Oreocharis benthamii in allopatry and sympatry based on ISSR data for 754 individuals from 26 populations, in order to understand the processes leading to speciation. Contrary to expectations, the facultative xenogamy (mixed mating) species O. benthamii has a relatively low genetic diversity within populations (H = 0.1014, I = 0.1528) and high genetic differentiation among populations (G ST = 0.5867, ФST = 0.659), as is typically found for selfing species. Genetic variance between the two varieties in sympatric populations (44%, ФST = 0.444) is significantly more than that in allopatric populations (14%, ФST = 0.138). Consistent with the taxonomical delimitation of the two varieties, all sampled individuals of O. benthamii clustered into two genetic groups. Moreover, the genetic structures of populations of both varieties are correlated with their different geographical origins. Our studies show that significant divergence between sympatric populations of the two varieties could be attributed primarily to reinforcement by genetic divergent selection in sympatry where secondary contact had occurred. The major proportion of the genetic variation in outcrossing and mixed mating plants may exist among populations when the populations are distributed in fragmented habitats, due to the paucity of suitable habitat combined with inefficient seed dispersal mechanism and limited pollinator foraging area that may limit the gene flow.


The development and codesign of the PATHway intervention: a theory-driven eHealth platform for the self-management of cardiovascular disease.
Cardiovascular diseases (CVDs) are a leading cause of premature death worldwide. International guidelines recommend routine delivery of all phases of cardiac rehabilitation (CR). Uptake of traditional CR remains suboptimal, as attendance at formal hospital-based CR programs is low, with community-based CR rates and individual long-term exercise maintenance even lower. Home-based CR programs have been shown to be equally effective in clinical and health-related quality of life outcomes and yet are not readily available. The aim of the current study was to develop the PATHway intervention (physical activity toward health) for the self-management of CVD. Increasing physical activity in individuals with CVD was the primary behavior. The PATHway intervention was theoretically informed by the behavior change wheel and social cognitive theory. All relevant intervention functions, behavior change techniques, and policy categories were identified and translated into intervention content. Furthermore, a person-centered approach was adopted involving an iterative codesign process and extensive user testing. Education, enablement, modeling, persuasion, training, and social restructuring were selected as appropriate intervention functions. Twenty-two behavior change techniques, linked to the six intervention functions and three policy categories, were identified for inclusion and translated into PATHway intervention content. This paper details the use of the behavior change wheel and social cognitive theory to develop an eHealth intervention for the self-management of CVD. The systematic and transparent development of the PATHway intervention will facilitate the evaluation of intervention effectiveness and future replication.


Complete Genome Sequence of Highly Adherent Pseudomonas aeruginosa Small-Colony Variant SCV20265.
The evolution of small-colony variants within Pseudomonas aeruginosa populations chronically infecting the cystic fibrosis lung is one example of the emergence of adapted subpopulations. Here, we present the complete genome sequence of the autoaggregative and hyperpiliated P. aeruginosa small-colony variant SCV20265, which was isolated from a cystic fibrosis (CF) patient.


Effects of dose and strain of vaccine on success of measles vaccination of infants aged 4-5 months.
Small-scale trials of the Edmonston-Zagreb (E-Z) measles vaccine were undertaken to determine the dose necessary to immunise 4-6-month-old infants. Antibody responses, measured 16 weeks after vaccination, were dose dependent: 40,000 plaque forming units given subcutaneously resulted in positive responses in all infants and higher antibody levels than doses of 20,000 or 10,000 units (10,000 units gave a failure rate of 25%). In further trials the E-Z vaccine was compared with the Schwarz vaccine, both being given in subcutaneous doses of 40,000 plaque forming units. In infants aged 20 weeks the E-Z vaccine produced higher levels of measles antibody and in those aged 18 weeks its superiority showed in a lower proportion failing to respond (3 of 39 versus 19 of 35).


Prolactinomas and nonfunctioning adenomas: preoperative diagnosis of tumor type using serum prolactin and tumor size.
Prolactinoma and nonfunctioning adenoma (NFA) are the most common sellar pathologies, and both can present with hyperprolactinemia. There are no definitive studies analyzing the relationship between the sizes of prolactinomas and NFAs and the serum prolactin level. Current guidelines for serum prolactin level cutoffs to distinguish between pathologies are suboptimal because they fail to consider the adenoma volume. In this study, the authors attempted to describe the relationship between serum prolactin level and prolactinoma volume. They also examined the predictive value that can be gained by considering tumor volume in differentiating prolactinoma from NFA and provide cutoff values based on a large sample of patients. A retrospective analysis of consecutive patients with prolactinomas (n = 76) and NFAs (n = 217) was performed. Patients were divided into groups based on adenoma volume, and the two pathologies were compared. A strong correlation was found between prolactinoma volume and serum prolactin level (r = 0.831, p < 0.001). However, there was no significant correlation between NFA volume and serum prolactin level (r = -0.020, p = 0.773). Receiver operating characteristic curve analysis of three different adenoma volume groups was performed and resulted in different serum prolactin level cutoffs for each group. For group 1 (≤ 0.5 cm3), the most accurate cutoff was 43.65 μg/L (area under the curve [AUC] = 0.951); for group 2 (> 0.5 to 4 cm3), 60.05 μg/L (AUC = 0.949); and for group 3 (> 4 cm3), 248.15 μg/L (AUC = 1.0). Prolactinoma volume has a significant impact on serum prolactin level, whereas NFA volume does not. This finding indicates that the amount of prolactin-producing tissue is a more important factor regarding serum prolactin level than absolute adenoma volume. Hence, volume should be a determining factor to distinguish between prolactinoma and NFA prior to surgery. Current serum prolactin threshold level guidelines are suboptimal and cannot be generalized across all adenoma volumes.


Evidence of an association between desmoglein 3 haplotypes and pemphigus vulgaris.
Pemphigus vulgaris (PV, OMIM 169610) is a severe blistering disorder of the skin and mucous membranes, caused by the production of autoantibodies directed against the epithelial adhesive protein desmoglein 3. Although an association between PV and HLA class II alleles has been established, the genetic factors predisposing to the disease remain poorly understood, the rarity of PV hampering the recruitment of substantial patient cohorts. To investigate DSG3 as a candidate PV susceptibility gene. We examined five DSG3 single nucleotide polymorphisms (rs8085532, rs3911655, rs3848485, rs3794925 and rs1466379) in two case-control datasets respectively originating from the U.K. (62 PV patients, 154 controls) and northern India (28 patients, 98 controls). In the U.K. sample, we observed a significant association between PV and the DSG3*TCCTC haplotype (Fisher's exact test P = 0.002). A related haplotype (DSG3*TCCCC) was associated with PV in the Indian dataset (P = 0.002). We also found that all British and Indian patients bearing DSG3 risk haplotypes carried at least one copy of a PV-associated HLA allele. These results suggest that genetic variation of DSG3 may be an additive risk factor predisposing to PV and warrant further investigations of this gene.


Direct and indirect effects of rotavirus vaccination: comparing predictions from transmission dynamic models.
Early observations from countries that have introduced rotavirus vaccination suggest that there may be indirect protection for unvaccinated individuals, but it is unclear whether these benefits will extend to the long term. Transmission dynamic models have attempted to quantify the indirect protection that might be expected from rotavirus vaccination in developed countries, but results have varied. To better understand the magnitude and sources of variability in model projections, we undertook a comparative analysis of transmission dynamic models for rotavirus. We fit five models to reported rotavirus gastroenteritis (RVGE) data from England and Wales, and evaluated outcomes for short- and long-term vaccination effects. All of our models reproduced the important features of rotavirus epidemics in England and Wales. Models predicted that during the initial year after vaccine introduction, incidence of severe RVGE would be reduced 1.8-2.9 times more than expected from the direct effects of the vaccine alone (28-50% at 90% coverage), but over a 5-year period following vaccine introduction severe RVGE would be reduced only by 1.1-1.7 times more than expected from the direct effects (54-90% at 90% coverage). Projections for the long-term reduction of severe RVGE ranged from a 55% reduction at full coverage to elimination with at least 80% coverage. Our models predicted short-term reductions in the incidence of RVGE that exceeded estimates of the direct effects, consistent with observations from the United States and other countries. Some of the models predicted that the short-term indirect benefits may be offset by a partial shifting of the burden of RVGE to older unvaccinated individuals. Nonetheless, even when such a shift occurs, the overall reduction in severe RVGE is considerable. Discrepancies among model predictions reflect uncertainties about age variation in the risk and reporting of RVGE, and the duration of natural and vaccine-induced immunity, highlighting important questions for future research.


A Mendelian locus on chromosome 16 determines susceptibility to doxorubicin nephropathy in the mouse.
The development of kidney disease is influenced by both genetic and environmental factors. Searching for models of glomerulopathy that display strong gene-environment interaction, we examined the determinants of anthracycline-induced nephropathy, a classic, strain-dependent experimental model applied to rodents in the past four decades. We produced three crosses derived from mice with contrasting susceptibility to doxorubicin (DOX) nephropathy and, surprisingly, we found that this widely studied model segregates as a single-gene defect with recessive inheritance. By genome-wide analysis of linkage, we mapped the trait locus to chromosome 16A1-B1 (DOXNPH locus) in all three crosses [peak logarithm of odds (lod) score of 92.7, P = 1 x 10(-65)]; this interval represents a susceptibility locus for nephropathy. Gene expression analysis indicated that susceptibility alleles at the DOXNPH locus are associated with blunted expression of protein arginine methyltransferase 7 (Prmt7) on chromosome 8, a protein previously implicated in cellular sensitivity to chemotherapeutic agents (lod = 12.4, P = 0.0001). Therefore, Prmt7 expression serves as a molecular marker for susceptibility to DOX nephropathy. Finally, increased variation in the severity of kidney disease among affected mice motivated a second genome-wide search, identifying a locus on chromosome 9 that influences the severity and progression of nephropathy (DOXmod, peak lod score 4.3, P = 0.0018). These data provide genetic and molecular characterization of a previously unrecognized Mendelian trait. Elucidation of DOX nephropathy may simultaneously provide insight into the pathogenesis of renal failure and mechanisms of cytotoxicity induced by chemotherapeutic agents.


The sensitization hypothesis and importance of psychosensory features in mood disorder: a review.
The sensitization model of mood disorders, articulated 10 years ago, predicts symptoms, course, and treatment response relationships. However, clinical data testing this model have not been reviewed. The authors reviewed all English-language publications characterizing psychosensory features (clinical phenomena hypothesized to reflect temporolimbic sensitization) and the relationship of these features to mood disorder, course, and treatment. They found that 20% to 80% of bipolar and 80% of epileptic patients experience psychosensory phenomena. Although these features do not identify a specific subgroup of mood disorders, their prevalence is highest in schizoaffective bipolar patients. The value of psychosensory features in predicting treatment response or chronicity is unclear.


Are pain and function better measures of outcome than revision rates after TKR in the younger patient?
Revision is the gold standard outcome measurement for survival analyses of orthopaedic implants but reliance on revision as an endpoint has been recently questioned. This study, that assesses long-term outcome in a specific group of patients who had undergone total knee replacement (TKR) for osteoarthritis, highlights the main problems facing modern survival analyses. Minimum 12-year survival and outcome data were reviewed for a series of sixty patients under the age of 60 years (mean age 55.4 years) who underwent total knee replacement (TKR) for osteoarthritis. The patients are a subgroup from a larger consecutive series of 1429 patients who underwent TKR between 1987 and 1993 at a single institution. Whilst the main study aim was to compare outcome of TKR using different endpoints, the outcome of TKR in this younger subpopulation could also be investigated. With revision as the primary endpoint the survival for TKR was 82.2% (95% CI 17.3). The mean OKS at follow-up (mean 15.7 years) was 30.9. However, many of the 82% of patients who did not undergo revision had a less than satisfactory outcome. 41% of these patients reported modest or severe pain (using the OKS) at final follow-up. A combined endpoint including revision, poor function and significant pain drastically reduced the survival rate for the operation. Survival based on revision alone provides an acceptable but inaccurate impression of outcome in younger TKR patients (under 60 years). A true representation of the success of TKR should include pain and function as endpoints.


Blunt versus penetrating subclavian artery injury: presentation, injury pattern, and outcome.
Subclavian artery (SCA) injuries are rare vascular injuries and may be difficult to manage. The majority of SCA injuries are secondary to penetrating trauma. The purpose of this report is to examine the injury patterns, diagnostic and therapeutic approaches, and outcome of patients with blunt and penetrating SCA injuries. Retrospective review Fifty-six patients sustained SCA injuries (25 blunt, 31 penetrating). SCA injury location was evenly distributed between the proximal, middle, and distal SCA after penetrating trauma; proximal injuries were rare (2 of 25) with blunt mechanisms. A radial arterial pulse deficit was present in only 3 of 25 blunt injuries and 9 of 31 penetrating injuries. Complications occurred more commonly in both groups of patients with initial systolic blood pressures less than 90 mm Hg. Survival was 76% in blunt and 81% in penetrating groups; limb salvage was similar (92% in blunt and 97% in penetrating groups). Complete brachial plexus injuries were more common with blunt injuries. SCA injuries are rare vascular injuries with an associated high morbidity and mortality, regardless of mechanism. Blunt mechanisms result in more middle and distal injuries and more frequent complete brachial plexus injuries. Complications are related to the hemodynamic status of the patient upon presentation, and not to mechanism of injury.


Shockwave treatment for medial tibial stress syndrome in athletes; a prospective controlled study.
Objective The purpose of this study was to describe the results of two treatment regimens for medial tibial stress syndrome (MTSS); a graded running programme and the same running programme with additional shockwave therapy (extracorporeal shockwave therapy; ESWT). Design A prospective observational controlled trial. Setting Two different sports medicine departments. Participants 42 athletes with MTSS were included. Intervention Patients from one hospital were treated with a graded running programme, while patients from the other hospital were treated with the same graded running programme and focused ESWT (five sessions in 9 weeks). Main Outcome Measures Time to full recovery (the endpoint was being able to run 18 min consecutively without pain at a fixed intensity). Results The time to full recovery was significantly faster in the ESWT group compared with the patients who only performed a graded running programme, respectively 59.7±25.8 and 91.6±43.0 days (p=0.008). Conclusions This prospective observational study showed that MTSS patients may benefit from ESWT in addition to a graded running programme. ESWT as an additional treatment warrants further investigation in a prospective controlled trial with the addition of randomisation and double blinding.


Genomic structure, expression and characterization of a STAT5 homologue from pufferfish (Tetraodon fluviatilis).
The STAT5 (signal transducer and activator of transcription 5) gene was isolated and characterized from a round-spotted pufferfish genomic library. This gene is composed of 19 exons spanning 11 kb. The full-length cDNA of Tetraodon fluviatilis STAT5 (TfSTAT5) contains 2461 bp and encodes a protein of 785 amino acid residues. From the amino acid sequence comparison, TfSTAT5 is most similar to mouse STAT5a and STAT5b with an overall identity of 76% and 78%, respectively, and has < 35% identity with other mammalian STATs. The exon/intron junctions of the TfSTAT5 gene were almost identical to those of mouse STAT5a and STAT5b genes, indicating that these genes are highly conserved at the levels of amino acid sequence and genomic structure. To understand better the biochemical properties of TfSTAT5, a chimeric STAT5 was generated by fusion of the kinase-catalytic domain of carp Janus kinase 1 (JAK1) to the C-terminal end of TfSTAT5. The fusion protein was expressed and tyrosine-phosphorylated by its kinase domain. The fusion protein exhibits specific DNA-binding and transactivation potential toward an artificial fish promoter as well as authentic mammalian promoters such as the beta-casein promoter and cytokine inducible SH2 containing protein (CIS) promoter when expressed in both fish and mammalian cells. However, TfSTAT5 could not induce the transcription of beta-casein promoter via rat prolactin and Nb2 prolactin receptor. To our knowledge, this is the first report describing detailed biochemical characterization of a STAT protein from fish.


Bi-allelic JAM2 Variants Lead to Early-Onset Recessive Primary Familial Brain Calcification.
Primary familial brain calcification (PFBC) is a rare neurodegenerative disorder characterized by a combination of neurological, psychiatric, and cognitive decline associated with calcium deposition on brain imaging. To date, mutations in five genes have been linked to PFBC. However, more than 50% of individuals affected by PFBC have no molecular diagnosis. We report four unrelated families presenting with initial learning difficulties and seizures and later psychiatric symptoms, cerebellar ataxia, extrapyramidal signs, and extensive calcifications on brain imaging. Through a combination of homozygosity mapping and exome sequencing, we mapped this phenotype to chromosome 21q21.3 and identified bi-allelic variants in JAM2. JAM2 encodes for the junctional-adhesion-molecule-2, a key tight-junction protein in blood-brain-barrier permeability. We show that JAM2 variants lead to reduction of JAM2 mRNA expression and absence of JAM2 protein in patient's fibroblasts, consistent with a loss-of-function mechanism. We show that the human phenotype is replicated in the jam2 complete knockout mouse (jam2 KO). Furthermore, neuropathology of jam2 KO mouse showed prominent vacuolation in the cerebral cortex, thalamus, and cerebellum and particularly widespread vacuolation in the midbrain with reactive astrogliosis and neuronal density reduction. The regions of the human brain affected on neuroimaging are similar to the affected brain areas in the myorg PFBC null mouse. Along with JAM3 and OCLN, JAM2 is the third tight-junction gene in which bi-allelic variants are associated with brain calcification, suggesting that defective cell-to-cell adhesion and dysfunction of the movement of solutes through the paracellular spaces in the neurovascular unit is a key mechanism in CNS calcification.


Intermittent neonatal hypoxia elicits the upregulation of inflammatory-related genes in adult male rats through long-lasting programming effects.
The long-term effects of neonatal intermittent hypoxia (IH), an accepted model of apnea-induced hypoxia, are unclear. We have previously shown lasting "programming" effects on the HPA axis in adult rats exposed to neonatal IH. We hypothesized that neonatal rat exposure to IH will subsequently result in a heightened inflammatory state in the adult. Rat pups were exposed to normoxia (control) or six cycles of 5% IH or 10% IH over one hour daily from postnatal day 2-6. Plasma samples from blood obtained at 114 days of age were analyzed by assessing the capacity to induce transcription in a healthy peripheral blood mononuclear cell (PBMC) population and read using a high-density microarray. The analysis of plasma from adult rats previously exposed to neonatal 5% IH versus 10% IH resulted in 2579 significantly regulated genes including increased expression of Cxcl1, Cxcl2, Ccl3, Il1a, and Il1b. We conclude that neonatal exposure to intermittent hypoxia elicits a long-lasting programming effect in the adult resulting in an upregulation of inflammatory-related genes.


Thermally invariant dielectric coatings for micromirrors.
Thermal expansion-induced curvature becomes a major effect in micromirrors as the mirror diameter exceeds 100 microm. Such mirrors are used for optical switching, scanning, and many other applications. By using multilayer coatings instead of a single metal reflector, one can use the mechanical properties of the multilayer to create mirrors with zero curvature across temperature. We demonstrate the fabrication of such thermally invariant mirrors using dielectric coatings. A semianalytic model based on free-plate elastic theory is developed that uses empirical parameters in place of the true thermal expansion coefficients of the coating materials. Micromirrors are demonstrated that maintain their design curvature to within lambda/60 for lambda = 633 nm across an operating range from 21 degrees C to 58 degrees C.


Facilitating Sustainable Disaster Risk Reduction in Indigenous Communities: Reviving Indigenous Worldviews, Knowledge, and Practices through Two-Way Partnering.
The Sendai Framework of Action 2015-2030 calls for holistic Indigenous disaster risk reduction (DRR) research. Responding to this call, we synergized a holistic philosophical framework (comprising ecological systems theory, symbolic interactionism, and intersectionality) and social constructionist grounded theory and ethnography within a critical Indigenous research paradigm as a methodology for exploring how diverse individual and contextual factors influence DRR in a remote Indigenous community called Galiwinku, in the Northern Territory of Australia. Working together, Indigenous and non-Indigenous researchers collected stories in local languages using conversations and yarning circles with 20 community members, as well as participant observations. The stories were interpreted and analysed using social constructivist grounded theory analysis techniques. The findings were dialogued with over 50 community members. The findings deeply resonated with the community members, validating the trustworthiness and relevance of the findings. The grounded theory that emerged identified two themes. First, local Indigenous knowledge and practices strengthen Indigenous people and reduce the risks posed by natural hazards. More specifically, deep reciprocal relationships with country and ecological knowledge, strong kinship relations, Elder's wisdom and authority, women and men sharing power, and faith in a supreme power/God and Indigenous-led community organizations enable DRR. Second, colonizing practices weaken Indigenous people and increase the risks from natural hazards. Therefore, colonization, the imposition of Western culture, the government application of top-down approaches, infiltration in Indigenous governance systems, the use of fly-in/fly-out workers, scarcity of employment, restrictions on technical and higher education opportunities, and overcrowded housing that is culturally and climatically unsuitable undermine the DRR capability. Based on the findings, we propose a Community-Based DRR theory which proposes that facilitating sustainable Indigenous DRR in Australian Indigenous communities requires Indigenous and non-Indigenous partners to genuinely work together in two-directional and complementary ways.


Management of cardiovascular risk factors in people with diabetes in primary care: cross-sectional study.
Cardiovascular disease is the major cause of morbidity and mortality in people with diabetes. The management of cardiovascular risk factors in people with diabetes in primary care was compared with National Institute of Clinical Excellence guidelines. A cross-sectional study in 26 general practices, with a combined list size of 256,188 patients, participating in the Kent, Surrey and Sussex Primary Care Research Network. Primary outcomes were process of care measures. Analysis of general practice computer data on the management of 5980 patients with diabetes, of whom 86% were aged 45 years and over. The prevalence of diabetes was 2.0% in women and 2.6% in men, much lower than the estimated expected prevalence of 4.8% for women and 3.3% in men. Blood pressure was well recorded (96% in both sexes), cholesterol levels less well (79% of women, 84% of men). Hypertension (78% of women, 72% of men) was common. Twenty-one percent of women and 16% of men had a blood pressure above 160/100 mmHg, suggesting under use of antihypertensive therapy. Cholesterol levels were >or=5 mmol/l in 46% of women and 38% of men. Lipid-lowering drugs were prescribed in 38% of women and men. Aspirin was prescribed in 38% of women and 40% of men. There is an under-diagnosis of diabetes and an under-treatment of blood pressure and blood cholesterol, more marked in women than in men. There is scope for improved management within general practice, including addressing sex inequalities.


A single nucleotide polymorphism in the NCF1 gene leading to reduced oxidative burst is associated with systemic lupus erythematosus.
Ncf1 polymorphisms leading to low production of reactive oxygen species (ROS) are strongly associated with autoimmune diseases in animal models. The human NCF1 gene is very complex with both functional and non-functional gene copies and genotyping requires assays specific for functional NCF1 genes. We aimed at investigating association and function of the missense single nucleotide polymorphism (SNP), rs201802880 (here denoted NCF1-339) in NCF1 with systemic lupus erythematosus (SLE). We genotyped the NCF1-339 SNP in 973 Swedish patients with SLE and 1301 controls, using nested PCR and pyrosequencing. ROS production and gene expression of type 1 interferon-regulated genes were measured in isolated cells from subjects with different NCF1-339 genotypes. We found an increased frequency of the NCF1-339 T allele in patients with SLE, 11% compared with 4% in controls, OR 3.0, 95% CI 2.4 to 3.9, p=7.0×10-20. The NCF1-339 T allele reduced extracellular ROS production in neutrophils (p=0.004) and led to an increase expression of type 1 interferon-regulated genes. In addition, the NCF1-339 T allele was associated with a younger age at diagnosis of SLE; mean age 30.3 compared with 35.9, p=2.0×1-6. These results clearly demonstrate that a genetically controlled reduced production of ROS increases the risk of developing SLE and confirm the hypothesis that ROS regulate chronic autoimmune inflammatory diseases.


[Cocaine use and sudden death: excited delirium syndrome].
Three patients, men aged 30, 33 and 32 years respectively, died in police custody in Amsterdam, the Netherlands. According to autopsy reports, all patients died from excited delirium syndrome induced by cocaine use. Excited delirium is a state of mental and physiological hyperarousal commonly associated with the use of cocaine. Patients suffering from excited delirium can be extremely hostile and violent. If a physical struggle arises with police or medical personnel, the episode can end in death due to adrenergic overstimulation. Patients in acute excited states should be regarded as a medical emergency with a serious mortality risk, and should be transferred to the hospital rather than the police station. Medical and police staff should aim to reduce confrontation. When patients are too agitated to cooperate, benzodiazepines should be used for sedation of the patient. Beta blockers and antipsychotic medication should not be administered, as these medications can exacerbate the fatal cardiac effects seen in excited delirium.


Body ownership: When feeling and knowing diverge.
Individuals with the peculiar disturbance of 'overcompleteness' experience an intense desire to amputate one of their healthy limbs, describing a sense of disownership for it (Body Integrity Identity Disorder - BIID). This condition is similar to somatoparaphrenia, the acquired delusion that one's own limb belongs to someone else. In ten individuals with BIID, we measured skin conductance response to noxious stimuli, delivered to the accepted and non-accepted limb, touching the body part or simulating the contact (stimuli approach the body without contacting it), hypothesizing that these individuals have responses like somatoparaphrenic patients, who previously showed reduced pain anticipation, when the threat was directed to the disowned limb. We found reduced anticipatory response to stimuli approaching, but not contacting, the unwanted limb. Conversely, stimuli contacting the non-accepted body-part, induced stronger SCR than those contacting the healthy parts, suggesting that feeling of ownership is critically related to a proper processing of incoming threats.


Mechanisms involved in alpha6beta1-integrin-mediated Ca(2+) signalling.
Contact of Jurkat T-lymphocytes with the extracellular matrix (ECM) protein laminin resulted in long-lasting alpha6beta1-integrin-mediated Ca(2+) signalling. Both Ca(2+) release from thapsigargin-sensitive Ca(2+) stores and capacitative Ca(2+) entry via Ca(2+) channels sensitive to SKF 96365 constitute important parts of this process. Inhibition of alpha6beta1-integrin-mediated Ca(2+) signalling by (1) the src kinase inhibitor PP2, (2) the PLC inhibitor U73122, and (3) the cyclic adenosine diphosphoribose (cADPR) antagonist 7-deaza-8-Br-cADPR indicate the involvement of src tyrosine kinases and the Ca(2+)-releasing second messengers D-myo-inositol 1,4,5-trisphosphate (InsP3) and cADPR.


Go big or go home: A model-based assessment of general strategies to slow the spread of forest pests via infested firewood.
Invasive pests, such as emerald ash borer or Asian longhorn beetle, have been responsible for unprecedented ecological and economic damage in eastern North America. These and other wood-boring invasive insects can spread to new areas through human transport of untreated firewood. Behaviour, such as transport of firewood, is affected not only by immediate material benefits and costs, but also by social forces. Potential approaches to reduce the spread of wood-boring pests through firewood include raising awareness of the problem and increasing the social costs of the damages incurred by transporting firewood. In order to evaluate the efficacy of these measures, we create a coupled social-ecological model of firewood transport, pest spread, and social dynamics, on a geographical network of camper travel between recreational destinations. We also evaluate interventions aimed to slow the spread of invasive pests with untreated firewood, such as inspections at checkpoints to stop the movement of transported firewood and quarantine of high-risk locations. We find that public information and awareness programs can be effective only if the rate of spread of the pest between and within forested areas is slow. Direct intervention via inspections at checkpoints can only be successful if a high proportion of the infested firewood is intercepted. Patch quarantine is only effective if sufficiently many locations can be included in the quarantine and if the quarantine begins early. Our results indicate that the current, relatively low levels of public outreach activities and lack of adequate funding are likely to render inspections, quarantine and public outreach efforts ineffective.


Update on unethical use of placebos in randomised trials.
The most recent (Fifth) revision of the Declaration of Helsinki, adopted in October 2000 by the World Medical Association (WMA), reinforces the longstanding prohibition against offering placebo instead of effective therapy. The WMA left no doubt that if a beneficial treatment for a condition has already been recognised, it is unethical to offer placebo in place of such treatment to anyone in a study of the same condition. We have previously drawn attention to the discrepancy between the spirit of the Declaration and the common practice of using placebo controls in randomised trials even if effective treatment exists. Despite the mandates of the Declaration of Helsinki and concern from ethicists and scientists, the US Food and Drug Administration (FDA) continues to demand and defend placebo-controlled evidence of efficacy and safety for the development of many new pharmaceuticals, even if effective therapy exists. We suggest that the FDA's arguments defending their practice are insufficient to justify medical research that violates the Declaration of Helsinki.


Prognostic factors for inpatient functional recovery following total hip and knee arthroplasty: a systematic review.
Background and purpose - Essential for safe and timely hospital discharge, inpatient functional recovery following lower limb arthroplasty is also variable. A previous systematic review reported moderate and conflicting levels of evidence regarding patient-related predictors of inpatient recovery for primary total hip arthroplasty (THA). A systematic review of surgical prognostic factors for inpatient recovery following THA or total knee arthroplasty (TKA) is yet to be undertaken. We identified patient and surgical prognostic factors for inpatient functional recovery following THA and TKA; determined whether inpatient functional recovery varies between these procedures; and established whether validated outcome measures relevant to the patient's functional requirements for hospital discharge are routinely assessed.Patients and methods - Critical Appraisal Skills Programme checklists assessed methodological quality, and a best-evidence synthesis approach determined the levels of evidence supporting individual prognostic factors. PubMed, CINAHL, Embase, Scopus, and PEDro databases were searched from inception to May 2019. Included studies examined patient or surgical prognostic factors and a validated measure of post-operative function within 2 weeks of primary, unilateral THA or TKA.Results - Comorbidity status and preoperative function are supported by a strong level of evidence for TKA. For THA, no strong level of evidence was found for patient-related prognostic factors, and no surgical factors were independently prognostic for either arthroplasty site. Limited evidence supports fast-track protocols in the TKA population.Interpretation - Preoperative screening and optimization is recommended. Assessment of Enhanced Recovery Pathways using validated outcome measures appropriate for the early postoperative period is warranted.


Ultrastructural changes of the nodose ganglion cells following an intraneural injection of Ricinus communis agglutinin-60 into the vagus nerve in hamsters.
Virtually all the ganglion cells in the nodose ganglion in hamsters underwent rapid degeneration following an intraneural injection of RCA-60 into the vagus nerve in the cervical region. The earliest signs of neuronal degeneration were evident in animals which survived 5 days after the ricin application. A remarkable feature was the appearance of a variable number of granular dense bodies measuring 1-4 microns in diameter in the cytoplasm. They were composed of closely stacked cisternae which were continuous at the periphery with those of the rough endoplasmic reticulum. Associated with the membranous cisternae were large accumulations of glycogen. With longer survival time, these glycogen-membrane complexes appeared to disintegrate. Numerous vacuoles and neurofilaments accumulated in their vicinity. Satellite cells were activated between the 7th and 10th postoperative days. These penetrated deeply into the degenerating neurons dividing them into numerous fragments by their extensive cytoplasmic prolongations. The cytoplasmic fragments of the RCA-poisoned neurons eventually became necrotic and disintegrated in the satellite cells, suggesting a rapid mode of neuronophagia. The biosynthesis of acetylcholinesterase was inhibited by the ricin injected as shown by the drastic reduction of the enzyme activity in the rough endoplasmic reticulum and nuclear envelope. Some isolated ganglion cells apparently survived the RCA injection as shown by their occurrence in long surviving animals (30-90 days). A few of them displayed an enhanced density of their cytoplasm and neurites. It is postulated that this was induced by the RCA released from the RCA-poisoned neurons.


Submicrometric hypromellose acetate succinate particles as carrier for soy isoflavones extract with improved skin penetration performance.
In this study, hypromellose acetate succinate (HPMCAS) stable submicronic particles loaded with a soy isoflavones extract have been obtained by nano spray drying technology. HPMCAS has been used as excipient able to increase both stability and supersaturation levels of the active ingredients hence able to enhance skin penetration performance of genistein and daidzein. The influence of polymer/extract ratio as other process variables, on particle size, morphology and permeation performance, have been investigated. Particles in submicronic range (mean size around 550nm) and narrow size distribution with high encapsulation efficiency (up to 86%) were obtained. HPMCAS was able to improve amorphization of genistein during the atomization process and avoid recrystallization during storage, even in harsh environmental condition. Moreover, the enhanced affinity of the optimized formulations with aqueous media, strongly increased isoflavones penetration through membrane with diffusive properties well-correlated to human skin, up to 10-fold higher than pure soy isoflavones extract raw material.


Analytical quality goals for parathyroid hormone based on biological variation.
Measurement of parathyroid hormone (PTH) is central in the investigation of pathologies of bone and mineral ion metabolism. Knowledge of the biological variation of an analyte forms an essential part of evaluating a new analyte, enabling the objective assessment of changes in serial results and the utility of reference intervals, as well as establishing laboratory quality specifications. This study determined the biological variation of PTH in 20 healthy individuals, which was calculated according to the familiar methods outlined by Fraser and Harris. The within-subject variation was 25.3% and the between-subject variation was 43.4%. The critical difference for sequential values significant at p<0.05 was calculated as 72%. The within-subject variation forms a relatively small part of the reference interval, shown by the low index of individuality of 0.58. Objective analytical quality goals have also been established, revealing achievable optimum performance for imprecision of approximately 6%. The desirable analytical bias goal was approximately 12%. This study has objectively shown that the analytical precision of current instruments is being achieved contrary to the known problems surrounding the analytical bias for PTH assays. The limitations of using reference intervals for PTH, both in diagnoses and monitoring, are shown.


Suppression of the constitutive expression of cytochrome P-450 2C11 by cytokines and interferons in primary cultures of rat hepatocytes: comparison with induction of acute-phase genes and demonstration that CYP2C11 promoter sequences are involved in the suppressive response to interleukins 1 and 6.
Hepatic expression of various members of the cytochrome P-450 (CYP) superfamily is suppressed during inflammatory responses. We have shown that the specific expression of P-450 2C11 in male rat liver is suppressed transcriptionally by endotoxin treatment. To investigate the molecular mechanisms underlying this phenomenon, we studied the effects of the inflammatory cytokines interleukin (IL)-1, IL-6, tumor necrosis factor-alpha (TNF), interferon (IFN)-alpha, and IFN-gamma on the expression of P-450 2C11 and the mRNAs of two typical acute-phase protein genes, alpha 1-acid glycoprotein (AGP) and fibrinogen, in primary hepatocyte cultures. IL-1, IL-6, TNF, and IFN-alpha all suppressed P-450 2C11 mRNA, whereas IFN-gamma had no effect. IL-1 and TNF were more effective than IL-6 in the suppression of P-450 2C11 mRNA. Whereas IL-1 and IL-6 effects on P-450 2C11 were accompanied by induction of AGP and fibrinogen mRNAs, IFN-alpha and TNF treatments had no effects on AGP. The suppression of P-450 2C11 and the induction of AGP by IL-1 showed similar time courses. The combination of IL-1 and IL-6 showed additivity in suppression of P-450 2C11, at maximally effective concentrations of cytokines. The effects of IL-1 on P-450 2C11 and AGP expression were blocked by IL-1 receptor antagonist protein. We also studied the effects of IL-1 and IL-6 on the transient expression of chloramphenicol acetyl-transferase reporter gene constructs containing 200 or 1287 base pairs of the 5' flanking region of the CYP2C11 gene, transfected into primary hepatocytes. The chloramphenicol acetyltransferase activities in cells transfected with the 200-base pair construct were reduced to about 33% and 58% of control levels by treatment with IL-1 or IL-6, respectively, suggesting that sequences important for cytokine down-regulation lie within the proximal promoter region of the CYP2C11 gene.


Keypoint recognition using randomized trees.
In many 3D object-detection and pose-estimation problems, runtime performance is of critical importance. However, there usually is time to train the system, which we will show to be very useful. Assuming that several registered images of the target object are available, we developed a keypoint-based approach that is effective in this context by formulating wide-baseline matching of keypoints extracted from the input images to those found in the model images as a classification problem. This shifts much of the computational burden to a training phase, without sacrificing recognition performance. As a result, the resulting algorithm is robust, accurate, and fast-enough for frame-rate performance. This reduction in runtime computational complexity is our first contribution. Our second contribution is to show that, in this context, a simple and fast keypoint detector suffices to support detection and tracking even under large perspective and scale variations. While earlier methods require a detector that can be expected to produce very repeatable results, in general, which usually is very time-consuming, we simply find the most repeatable object keypoints for the specific target object during the training phase. We have incorporated these ideas into a real-time system that detects planar, nonplanar, and deformable objects. It then estimates the pose of the rigid ones and the deformations of the others.


Demographic and lifestyle predictors of body mass index among offshore oil industry workers: cross-sectional and longitudinal findings.
Significant overweight among offshore workers on North Sea oil and gas installations has been linked to high calorie intake, lack of active leisure-time pursuits, and environmental factors conducive to weight gain. However, the prevalence of overweight among offshore workers has not been examined in recent data, and no longitudinal studies of body mass index (BMI) in this occupational group have been reported. Aims The present study sought to examine BMI levels in a sample of UK offshore personnel, and to evaluate demographic factors, smoking and work-related physical activity as predictors of BMI, and 5 year change in BMI. Survey data (including age, education, marital status, work-related physical activity and height/weight) were collected in 1995 from male workers on 17 North Sea installations (n = 1581, 83% response rate); follow-up data were obtained in 2000 (n = 354, 34.9% of the potential sample). Overall mean BMI was 25.6 (2.8) kg/m(2): rates of obesity (BMI > 30) and overweight (BMI = 25-30) were 7.5 and 47.3%, respectively. Mean age was 38.7 (8.9) years; linear and quadratic age terms predicted BMI. Age-adjusted BMI values were very similar to those reported from other offshore studies over the past 15 years. Age, marital status, education, smoking and physical activity significantly predicted baseline BMI, but only age (and some interactive effects) predicted 5 year BMI change. The present age-adjusted BMI values were closely similar to those found offshore in the mid-1980s, but also to recent national data; thus, North Sea personnel do not appear to reflect current population trends towards increased BMI levels. This result accords with the emphasis now given to health promotion (particularly dietary change) on offshore installations; the present findings also highlight the need to focus these initiatives on workers with sedentary jobs and/or low education.


Eye blinks are related to auditory information processing: evidence from a complex speech perception task.
There is increasing evidence that spontaneous eye blinks are related to mental states and can predict performance in certain tasks because of their relation to dopaminergic activity. Moreover, it has been shown that eye blinks while performing visual tasks are preferably executed not before all available information and even the manual response has been processed and given. Thus, blinks provide a natural endpoint of visual information processing. In the present study, we investigate to what degree such functional assignment of eye blinks also applies when only auditory stimuli are processed. For that, we present blink analyses on data of an auditory stock price monitoring task to examine the timing and frequency of blinks relative to the temporal dynamics of the task and different kinds of available cues. Our results show that blinks are meaningfully rather than randomly paced, although no visual information has to be processed. Blinks are significantly accelerated if a no-go trial is indicated which made all the subsequent information irrelevant. Although the stimuli were exclusively auditory, blinks were mostly inhibited during stimulus presentation. Taken together, blinks depend on the information being presented and mark a distinct point in time at which this information is conclusively processed. These findings deliver further support for the usefulness of eyeblink analyses, independently of the modality of the information being processed.


Cost-effectiveness analysis of fidaxomicin versus vancomycin in Clostridium difficile infection.
Fidaxomicin was non-inferior to vancomycin with respect to clinical cure rates in the treatment of Clostridium difficile infections (CDIs) in two Phase III trials, but was associated with significantly fewer recurrences than vancomycin. This economic analysis investigated the cost-effectiveness of fidaxomicin compared with vancomycin in patients with severe CDI and in patients with their first CDI recurrence. A 1 year time horizon Markov model with seven health states was developed from the perspective of Scottish public healthcare providers. Model inputs for effectiveness, resource use, direct costs and utilities were obtained from published sources and a Scottish expert panel. The main model outcome was the incremental cost-effectiveness ratio (ICER), expressed as cost per quality-adjusted life year (QALY), for fidaxomicin versus vancomycin; ICERs were interpreted using willingness-to-pay thresholds of £20,000/QALY and £30,000/QALY. One-way and probabilistic sensitivity analyses were performed. Total costs were similar with fidaxomicin and vancomycin in patients with severe CDI (£14,515 and £14,344, respectively) and in patients with a first recurrence (£16,535 and £16,926, respectively). Improvements in clinical outcomes with fidaxomicin resulted in small QALY gains versus vancomycin (severe CDI, +0.010; patients with first recurrence, +0.019). Fidaxomicin was cost-effective in severe CDI (ICER £16,529/QALY) and dominant (i.e. more effective and less costly) in patients with a first recurrence. The probability that fidaxomicin was cost-effective at a willingness-to-pay threshold of £30,000/QALY was 60% for severe CDI and 68% in a first recurrence. Fidaxomicin is cost-effective in patients with severe CDI and in patients with a first CDI recurrence versus vancomycin.


Replication of the Stockholm Adoption Study of alcoholism. Confirmatory cross-fostering analysis.
Two forms of alcoholism with distinct clinical features and mode of inheritance were first distinguished in the Stockholm Adoption Study. This involved a large sample of children born in Stockholm, Sweden, who were adopted at an early age and reared by nonrelatives. Type 1 alcoholism had adult onset and rapid progression of dependence without criminality, whereas type 2 had teenage onset of recurrent social and legal problems from alcohol abuse. A replication study was carried out with 577 men and 660 women born in Gothenburg, Sweden, and adopted at an early age/by nonrelatives. The genetic and environmental backgrounds of the adoptees were classified by the exact procedures calibrated by discriminant analysis in the original study. Both type 2 and severe type 1 alcoholism were confirmed as independently heritable forms of alcoholism in male adoptees. The lifetime risk of severe alcoholism was increased 4-fold in adopted men with both genetic and environmental risk factors characteristic of type 1 alcoholism compared with the others (11.4% vs 3.0%). Neither genetic nor environmental risk factors for type 1 alcoholism by themselves were sufficient to cause alcoholism. In contrast, the risk of type 2 alcoholism was increased 6-fold in adopted sons with a type 2 genetic background compared with others; regardless of their postnatal environment (10.7% vs 2.0%). The sons with a type 2 genetic background in the replication sample had no excess of type 1 alcoholism, and vice versa. There was no increased risk of mild abuse in adopted men regardless of their genetic or environmental background. Type 1 and type 2 alcoholism are clinically distinct forms of alcoholism with causes that are independent but not mutually exclusive.


Visualization of Toxoplasma gondii stage conversion by expression of stage-specific dual fluorescent proteins.
To recognize the stage conversion of Toxoplasma gondii between tachyzoite and bradyzoite in live host cells, a transgenic T. gondii line, which expressed stage-specific red and green fluorescence, was constructed. T. gondii PLK strain tachyzoites were stably transformed with genes encoding red fluorescent protein (DsRed Express) and green fluorescent protein (GFP) under the control of tachyzoite-specific SAG1 and bradyzoite-specific BAG1 promoters, respectively. The resulting transgenic parasite was designated PLK/DUAL. When PLK/DUAL was cultured in pH 7.0 medium, the PLK/DUAL zoites expressed red fluorescence, but no detectable levels of green fluorescence were observed. The PLK/DUAL zoites reacted with anti-SAG1 antibody, but not anti-BAG1 antiserum. When PLK/DUAL was cultured under high pH conditions, or in the presence of the p38 MAPK inhibitor SB202190, a small number of zoites expressed green fluorescence and were BAG1 positive. C57BL/6J mice were infected with PLK/DUAL tachyzoites. During the acute and reactivating phase, zoites expressed red fluorescence. However, green fluorescence was not detectable. By contrast, latent cysts expressed green fluorescence. The stage-specific dual fluorescence of PLK/DUAL facilitates identification of the parasitic stage in live cells, with the advantage that fixation or immunostaining is not required.


The Golgin Protein Giantin Regulates Interconnections Between Golgi Stacks.
Golgins are a family of Golgi-localized long coiled-coil proteins. The major golgin function is thought to be the tethering of vesicles, membranes, and cytoskeletal elements to the Golgi. We previously showed that knockdown of one of the longest golgins, Giantin, altered the glycosylation patterns of cell surfaces and the kinetics of cargo transport, suggesting that Giantin maintains correct glycosylation through slowing down transport within the Golgi. Giantin knockdown also altered the sizes and numbers of mini Golgi stacks generated by microtubule de-polymerization, suggesting that it maintains the independence of individual Golgi stacks. Therefore, it is presumed that Golgi stacks lose their independence following Giantin knockdown, allowing easier and possibly increased transport among stacks and abnormal glycosylation. To gain structural insights into the independence of Golgi stacks, we herein performed electron tomography and 3D modeling of Golgi stacks in Giantin knockdown cells. Compared with control cells, Giantin-knockdown cells had fewer and smaller fenestrae within each cisterna. This was supported by data showing that the diffusion rate of Golgi membrane proteins is faster in Giantin-knockdown Golgi, indicating that Giantin knockdown structurally and functionally increases connectivity among Golgi cisternae and stacks. This increased connectivity suggests that contrary to the cis-golgin tether model, Giantin instead inhibits the tether and fusion of nearby Golgi cisternae and stacks, resulting in transport difficulties between stacks that may enable the correct glycosylation of proteins and lipids passing through the Golgi.