Update on Minimalist Running Shoes.
This article provides updated information comparing minimalist running shoes (MRS) to conventional running shoes (CRS). Our previous review found that, compared with running in CRS, transitioning to MRS resulted in lower energy cost and less ground contact occurring at the forefoot, resulting in lower impact forces. There was some increased risk of injury with MRS, although data were conflicting. A more recent 26-week study involved 30 trained runners using CRS and 31 using MRS. The proportion of training time in the assigned shoes increased by 5% each week. After the first 6 weeks of transition (35% of training time in the assigned shoe), energy cost was lower and 5-km running time faster in MRS compared with CRS. No further improvement occurred from weeks 6 to 26. There were no significant differences in injury incidence in the two groups (CRS = 37%, MRS = 52%; p = .24). Running-related pain was higher in the MRS group in the knee, shin, calf, and ankle and increased at these locations as running mileage increased. Risk of injury in MRS increased as participant body weight increased. These more recent data suggest that MRS can improve performance, but most runners should limit running in MRS to 35% of training time and in situations where optimal performance is desired (e.g., races, fitness tests).


A novel subnetwork alignment approach predicts new components of the cell cycle regulatory apparatus in Plasmodium falciparum.
According to the World Health organization, half the world's population is at risk of contracting malaria. They estimated that in 2010 there were 219 million cases of malaria, resulting in 660,000 deaths and an enormous economic burden on the countries where malaria is endemic. The adoption of various high-throughput genomics-based techniques by malaria researchers has meant that new avenues to the study of this disease are being explored and new targets for controlling the disease are being developed. Here, we apply a novel neighborhood subnetwork alignment approach to identify the interacting elements that help regulate the cell cycle of the malaria parasite Plasmodium falciparum. Our novel subnetwork alignment approach was used to compare networks in Escherichia coli and P. falciparum. Some 574 P. falciparum proteins were revealed as functional orthologs of known cell cycle proteins in E. coli. Over one third of these predicted functional orthologs were annotated as "conserved Plasmodium proteins" or "putative uncharacterized proteins" of unknown function. The predicted functionalities included cyclins, kinases, surface antigens, transcriptional regulators and various functions related to DNA replication, repair and cell division. The results of our analysis demonstrate the power of our subnetwork alignment approach to assign functionality to previously unannotated proteins. Here, the focus was on proteins involved in cell cycle regulation. These proteins are involved in the control of diverse aspects of the parasite lifecycle and of important aspects of pathogenesis.


Compassion: an evolutionary analysis and empirical review.
What is compassion? And how did it evolve? In this review, we integrate 3 evolutionary arguments that converge on the hypothesis that compassion evolved as a distinct affective experience whose primary function is to facilitate cooperation and protection of the weak and those who suffer. Our empirical review reveals compassion to have distinct appraisal processes attuned to undeserved suffering; distinct signaling behavior related to caregiving patterns of touch, posture, and vocalization; and a phenomenological experience and physiological response that orients the individual to social approach. This response profile of compassion differs from those of distress, sadness, and love, suggesting that compassion is indeed a distinct emotion. We conclude by considering how compassion shapes moral judgment and action, how it varies across different cultures, and how it may engage specific patterns of neural activation, as well as emerging directions of research.


An analysis of graft-versus-host disease in Syrian hamsters. II. The epidermolytic syndrome: studies on its pathogenesis.
The epidermolytic syndrome that can be obtained at will in F(1) hybrid hamsters by the cutaneous inoculation of adequate doses of parental strain lymphoid cells has been investigated to determine whether the cutaneous lesions are due to an autoimmune process arising from the severe, initial GVH reactions in the skin. It was amply demonstrated that inoculation of donor cells into the skin was of crucial importance to the development of epidermolysis. Parental strain lymphoid cells in similar doses delivered by any other route into normal F(1) hybrids failed absolutely to incite the acute syndrome. If "immune lymphocyte transfer" reactions incited by donor cells in the host's skin were surgically removed at timed intervals after inoculation, only complete excision within 24 hr prevented the appearance of epidermolysis in F(1) hybrid hosts, indicating that inoculated donor cells must remain within the confines of the skin for approximately 24 hr in order to evoke the disease, persistence for longer periods of time being unnecessary for the subsequent course of the disease. However, reconstitution experiments involving the intramuscular inoculation of suspensions containing mixtures of donor cells and host lymphoid cells, in the presence or absence of epidermal cells, unequivocally indicated that no intimate exposure of lymphoid cells to putative skin-specific antigens was essential. Similarly, the elicitation of generalized epidermolysis in F(1) hybrids irradiated with 300 r and then inoculated intravenously with donor cells casts further doubt on the pathogenic importance of the skin as a source of tissue-specific antigen. The results of subsequent experiments indicated that host leukocytes, rather than parenchymal cells of the dermis or the epidermis, were important contributors of the transplantation antigenic stimulus. Moreover, a series of experiments, using (CB x MHA)F(1) hybrid hosts that had been lethally irradiated and reconstituted with bone marrow cells from ALS-treated MHA donors, indicated that from 6 to 10 wk after reconstitution-when direct and immune lymphocyte transfer reactions showed a virtual absence of native F(1) leukocytes from the circulation-donor cells obtained from specifically sensitized MHA donors were completely ineffective in inducing epidermolysis, while equivalent lymphoid cell inocula derived from CB donors evoked the cutaneous disease irrespective of the time elapsed since reconstitution. To explain these findings it is postulated that in hamsters, the primary targets in graft-versus-host disease incited by the intracutaneous inoculation of donor cells are leukocytes originating in bone marrow or lymph node, or both.


Murine IL-10 gene transfer inhibits established collagen-induced arthritis and reduces adenovirus-mediated inflammatory responses in mouse liver.
The effects of homologous IL-10 administration during an established autoimmune disease are controversial, given its reported immunostimulatory and immunosuppressive properties. Studies of collagen-induced arthritis have shown efficacy with repeated administrations of IL-10; however, when the EBV IL-10 homologue was administered via adenovirus gene transfer technology the results were equivocal. Therefore, the present study was undertaken to elucidate the effects of prolonged homologous IL-10 administration via adenovirus-mediated gene delivery on the progression of established arthritis. Collagen type II (CII)-immunized mice received i.v. injections of 10(7) or 10(8) PFU of an E1-deleted adenoviral vector containing the murine IL-10 gene (AdIL-10), after arthritis onset. Mice were monitored for 3 wk for disease progression, and gene transduction was assessed by quantification of serum mIL-10. CII-specific cell-mediated and humoral immune responses were analyzed by lymph node cell proliferation, cytokine production, and anti-CII Ab responses. Furthermore, because adenoviral vectors have been reported to induce organ dysfunction due to cell-mediated immune responses to the viral Ags, we have also evaluated delayed-type hypersensitivity responses and reactive hepatitis to the systemically delivered adenovirus and whether the IL-10 produced could influence those responses. Sustained suppression of autoimmune arthritis and elevated serum levels of IL-10 were achieved in our study. AdIL-10 treatment reduced cell-mediated immune reactivity, but did not affect humoral responses. Furthermore, IL-10 was able to reduce, but not totally abrogate, adenovirus-induced hepatic inflammation. These findings provide further insights into the diverse interplay of immune processes involved in autoimmune inflammation and the mechanism of cytokine immunotherapy.


Media content analysis of the introduction of a "soft opt-out" system of organ donation in Wales 2015-17.
In an attempt to improve organ donation rates, some countries are considering moving from "opt-in" systems where citizens must express their willingness to be an organ donor, to "opt-out" systems where consent is presumed unless individuals have expressed their wishes otherwise, by, for example, joining an "opt-out" register. In Wales-a part of the United Kingdom-the devolved government recently legislated for an "opt-out" system. For the change to be effective, a public awareness campaign was critical to the policy's success. Using quantitative and qualitative content analysis, we explored media coverage of the change to better understand the relationship between the state, policy actors, media and the public when such policy changes take place. Our findings illustrate how a state communication campaign can effectively set the media agenda within which we saw a degree of interdependency created with the state using the media to promote policy, and the media relying on the state for credible information. Yet we also found that the media is not uncritical and observed how it uses its autonomy to influence policy setting. Over the period of study, we found that a change in tone and view towards deemed consent organ donation has taken place in the media. However, while this may influence or reflect public attitudes, it is yet to be seen whether the media campaign translates into behavioural change that will result in increases in organ donations.


Healthcare-associated Pneumonia and Aspiration Pneumonia.
Healthcare-associated pneumonia (HCAP) is a new concept of pneumonia proposed by the American Thoracic Society/Infectious Diseases Society of America in 2005. This category is located between community-acquired pneumonia and hospital-acquired pneumonia with respect to the characteristics of the causative pathogens and mortality, and primarily targets elderly patients in healthcare facilities. Aspiration among such patients is recognized to be a primary mechanism for the development of pneumonia, particularly since the HCAP guidelines were published. However, it is difficult to manage patients with aspiration pneumonia because the definition of the condition is unclear, and the treatment is associated with ethical aspects. This review focused on the definition, prevalence and role of aspiration pneumonia as a prognostic factor in published studies of HCAP and attempted to identify problems associated with the concept of aspiration pneumonia.


Oriented mentalization-based treatment for borderline personality disorder patients: preliminary results at Camposampiero Mental Health Center.
This contribution presents two brief reports about preliminary results of 18 months of oriented Mentalization-Based Treatment (MBT), with Borderline Personality Disorder patients, recruited at the Camposampiero Mental Health Center. Following in large part Bateman and Fonagy guidelines for MBT in institutional settings, this paper presents preliminary results in two brief reports related to two cohorts of patient underwent to the oriented MBT in Camposampiero Mental Health Center (MHC). In the first study, we analyzed a group of 9 patients: an anamnestic schedule was administered; then, symptoms (SCL-90-R), psychodiagnostic scale and global health functioning (Health of the Nation Outcome Scale, Structured Clinical Interview for DSM-IV Axis II Disorder [SCID-II], Global Assessment of Functioning, [GAF]), data on service impact and service costs (Cassel Community Adjustment Questionnaire, Patient Evaluation Schedule and folder data) were provided at the beginning (T0), at the end of the treatment (T2) and 1 year after the end of the MBT project. The second study showed a micro-analytical change on a second patient cohort (n=6) at T0, 3, 6 and 9 months (T1) were presented considering specifically mentalization (Comparative Psychotherapy Process Scale, Modes of Mentalization Scale, Mentalization Imbalances Scale) and patienttherapist session evaluation trends (Session Evaluation Questionnaire) and patient reflective functioning at T0 and T2 (Reflective Functioning Questionnaire). Aims and hypotheses of the two studies pointed out how the oriented MBT bring to an improvement of the overall functioning of the patients, a reduction of the symptoms, a decrease of the diagnostic criteria for the Borderline Personality Disorder (BPD) and the other Axis II disorders, a reduction of the workload hours for the MHC staff and of the costs of the assistance. The analyses of both the studies were carried out using non-parametric statistics (Friedman test, Spearman correlation, Chi-square). Preliminary results confirmed the improvement in the overall functioning of patients (GAF), the reduction in BPD-related symptoms and in diagnostic criteria for BPD (SCID-II), the improvement of patients' mentalization skills, and a significant reduction in workload for health staff. Standing the limits and the preliminary results, the two brief reports demonstrate the feasibility of an oriented MBT within an Italian Public Service and the effectiveness of this treatment pathway for patients with BPD, leaving some open questions to stimulate a fruitful clinical discussion.


Clinical application of fish-oil intravenous lipid emulsion in adult home parenteral nutrition patients.
High-ω-6 polyunsaturated fatty acids (PUFAs) are noted to contribute to development of intestinal failure-associated liver disease (IFALD) in home parenteral nutrition (HPN). Fish oil (FO) has been added to latest generation of lipid injectable emulsion (ILE) to increase ω-3:ω-6 PUFA ratio; however, appropriate dose of FO to treat IFALD is unknown. After approval of exclusive FO ILE in the US for pediatric patients, we noted 2 adult patients with ongoing IFALD despite transition to mixed-oil (MO) ILE. They were transitioned to off-label FO ILE after review of literature regarding use of FO ILE in adult HPN patients was conducted to guide management. The first case involves a 40-year-old female receiving HPN with IFALD refractory to MO ILE. MO ILE (with 15% FO) was provided at 50 g/d for 3 d/wk and combined with FO ILE at 50 g/d for 4 d/wk. This combination resulted in improvement in liver studies and allowed for decrease in dextrose calories. The second case involves a 49-year-old male receiving HPN (secondary to complications of necrotizing pancreatitis) who developed IFALD. FO ILE was used as the sole source of lipids and led to improvement in liver function tests. No evidence of essential fatty acid deficiency was found in either case. Current case presentations and review of literature support the use of FO ILE to increase ω-3 PUFAs in patients with IFALD refractory to MO ILE. Additional research is necessary to delineate the dose of FO ILE necessary to achieve benefit.


Serum cholesterol in treatment-resistant depression.
Patients with major depressive disorder (MDD) may have significant differences in cholesterol levels compared with healthy controls. A previous study by our group reported that depressed patients with elevated cholesterol levels (>or=200 mg/dl) were significantly more likely to be nonresponders to fluoxetine treatment than depressed patients with nonelevated cholesterol levels. However, very little is known regarding cholesterol in patients with treatment-resistant depression (TRD). The purpose of this study was to compare cholesterol levels at baseline between depressed patients with and without TRD and to test whether cholesterol levels at baseline can predict clinical response in patients with TRD treated with open-label nortriptyline (NT). Ninety-two patients with TRD entered a 6-week open trial of NT. Baseline cholesterol levels were randomly collected for 59 of these patients. Controlling for age and gender, we compared baseline cholesterol and triglyceride levels for 35 patients with TRD who did not respond to NT with 205 non-TRD patients who responded to an 8-week open trial of fluoxetine. Furthermore, with the use of logistic regression, we tested whether baseline cholesterol levels predicted clinical response to NT in the patients with TRD. Patients with TRD had higher triglyceride levels at baseline compared with depressed patients without TRD. Cholesterol defined as a dichotomous variable being elevated if equal to or greater than 200 mg/dl, predicted poor response to a 6-week open trial of NT in patients with TRD. The results of this study confirm the relationship between hypercholesterolemia and poor outcome in the treatment of MDD for patients with TRD.


Fast and accurate prediction of the regioselectivity of electrophilic aromatic substitution reactions.
While computational prediction of chemical reactivity is possible it usually requires expert knowledge and there are relatively few computational tools that can be used by a bench chemist to help guide synthesis. The RegioSQM method for predicting the regioselectivity of electrophilic aromatic substitution reactions of heteroaromatic systems is presented in this paper. RegioSQM protonates all aromatic C-H carbon atoms and identifies those with the lowest free energies in chloroform using the PM3 semiempirical method as the most nucleophilic center. These positions are found to correlate qualitatively with the regiochemical outcome in a retrospective analysis of 96% of more than 525 literature examples of electrophilic aromatic halogenation reactions. The method is automated and requires only a SMILES string of the molecule of interest, which can easily be generated using chemical drawing programs such as ChemDraw. The computational cost is 1-10 minutes per molecule depending on size, using relatively modest computational resources and the method is freely available via a web server at ; http://www.regiosqm.org. RegioSQM should therefore be of practical use in the planning of organic synthesis.


Signals from dying hepatocytes trigger growth of liver progenitors.
The death rate of mature hepatocytes is chronically increased in various liver diseases, triggering responses that prevent liver atrophy, but often cause fibrosis. Mice with targeted disruption of inhibitor kappa B kinase (Ikk) in hepatocytes (HEP mice) provide a model to investigate this process because inhibiting Ikk-nuclear factor-kappaB (NF-kappaB) signalling in hepatocytes increases their apoptosis. Cell proliferation, apoptosis, progenitors, fibrosis and production of Hedgehog (Hh) ligands (progenitor and myofibroblast growth factors) were compared in HEP and control mice before and after feeding methionine choline-deficient ethionine-supplemented (MCDE) diets. Ikkbeta was deleted from primary hepatocytes to determine the effects on Hh ligand production; Hh signalling was inhibited directly in progenitors to determine the effects on viability. Liver sections from patients were examined to assess relationships between hepatocyte production of Hh ligands, accumulation of myofibroblastic cells and liver fibrosis. Disrupting the Ikk-NF-kappaB pathway in hepatocytes inhibited their proliferation but induced their production of Hh ligands. The latter provided viability signals for progenitors and myofibroblasts, enhancing accumulation of these cell types and causing fibrogenesis. Findings in the mouse models were recapitulated in diseased human livers. Dying mature hepatocytes produce Hh ligands which promote the compensatory outgrowth of progenitors and myofibroblasts. These results help to explain why diseases that chronically increase hepatocyte death promote cirrhosis.


Measurement of rRNA Variations in Natural Communities of Microorganisms on the Southeastern U.S. Continental Shelf.
The development of a clear understanding of the physiology of marine prokaryotes is complicated by the difficulties inherent in resolving the activity of various components of natural microbial communities. Application of appropriate molecular biological techniques offers a means of overcoming some of these problems. In this regard, we have used direct probing of bulk RNA purified from selective size fractions to examine variations in the rRNA content of heterotrophic communities and Synechococcus populations on the southeastern U.S. continental shelf. Heterotrophic communities in natural seawater cultures amended with selected substrates were examined. Synechococcus populations were isolated from the water column by differential filtration. The total cellular rRNA content of the target populations was assayed by probing RNA purified from these samples with an oligonucleotide complementing a universally conserved region in the eubacterial 16S rRNA (heterotrophs) or with a 1.5-kbp fragment encoding the Synechococcus sp. strain WH 7803 16S rRNA (cyanobacteria). The analyses revealed that heterotrophic bacteria responded to the addition of glucose and trace nutrients after a 6-h lag period. However, no response was detected after amino acids were added. The cellular rRNA content increased 48-fold before dropping to a value 20 times that detected before nutrients were added. Variations in the rRNA content from Synechococcus spp. followed a distinct diel pattern imposed by the phasing of cell division within the irradiance cycle. The results indicate that careful application of these appropriate molecular biological techniques can be of great use in discerning basic physiological characteristics of selected natural populations and the mechanisms which regulate growth at the subcellular level.


Ultrasensitive refractive index sensor based on the resonant scattering effect between double air circular-holes on silicon waveguides.
Recently, we have proposed a sensitive refractive index sensor design by integrating a circular-hole defect with an etched diffraction grating (EDG) spectrometer based on amorphous silicon photonic platforms. In the present paper, we will show that a much better sensitivity (~17422 nm/RIU) can be obtained by using double circular-holes with an appropriate interval. The influence of the double-hole interval on the performance of sensing applications is also characterized. A sinusoidal pattern of the sensitivity can be found as the interval increases. However, the intensity of the resonant peak (i.e., the detectability for sensing applications) significantly oscillates as the interval varies.


Postnatal estradiol up-regulates lung nitric oxide synthases and improves lung function in bronchopulmonary dysplasia.
Nitric oxide (NO) plays an important role in lung development and perinatal lung function, and pulmonary NO synthases (NOS) are decreased in bronchopulmonary dysplasia (BPD) following preterm birth. Fetal estradiol levels increase during late gestation and estradiol up-regulates NOS, suggesting that after preterm birth estradiol deprivation causes attenuated lung NOS resulting in impaired pulmonary function. To test the effects of postnatal estradiol administration in a primate model of BPD over 14 days after delivery at 125 days of gestation (term = 185 d). Cardiopulmonary function was assessed by echocardiography and whole body plethysmography. Lung morphometric and histopathologic analyses were performed, and NOS enzymatic activity and abundance were measured. Estradiol caused an increase in blood pressure and ductus arteriosus closure. Expiratory resistance and lung compliance were also improved, and this occurred before spontaneous ductal closure. Furthermore, both oxygenation and ventilation indices were improved with estradiol, and the changes in lung function and ventilatory support requirements persisted throughout the study period. Whereas estradiol had negligible effect on indicators of lung inflammation and on lung structure assessed after the initial 14 days of ventilatory support, it caused an increase in lung neuronal and endothelial NOS enzymatic activity. In a primate model of BPD, postnatal estradiol treatment had favorable cardiovascular impact, enhanced pulmonary function, and lowered requirements for ventilatory support in association with an up-regulation of lung NOS. Estradiol may be an efficacious postnatal therapy to improve lung function and outcome in preterm infants.


Are chronotype, social jetlag and sleep duration associated with health measured by Work Ability Index?
The present study investigates the impact of chronotype, social jetlag and sleep duration on self-perceived health, measured by Work Ability Index (WAI), within an industrial setting. Between 2011 and 2013, 2474 day and shift workers participated in a health check offered by an occupational health promotion program and filled out the Munich ChronoType Questionnaire (adapted to the rotational 12-h schedule for shift workers) and the WAI. We computed sleep duration on work and free days, chronotype, and social jetlag. We used linear regression models to examine chronotype, sleep duration and social jetlag for association with the WAI sum score, and proportional odds models to estimate the combined effect of social jetlag and sleep duration. Participants reported an average daily sleep duration of 7.35 h (SD: 1.2 h), had an average chronotype of 3:08 a.m. (SD: 1 h), and the average social jetlag corresponded to 1.96 h (SD: 2.05 h). Increasing social jetlag and shorter sleep duration were independently associated with a decreasing WAI, while chronotype per se was not associated with WAI. Short sleep duration combined with high social jetlag significantly increased the risk of poor WAI (OR = 1.36; 95% CI: 1.09-1.72), while long sleep duration and high social jetlag were not associated with poor WAI (OR = 1.09; 95% CI: 0.88-1.35). Our results add to a growing body of literature, suggesting that circadian misalignment, but not chronotype per se, may be critical for health. Our results indicate that longer sleep may override the adverse effects of social jetlag on WAI.


Identification and Risk Assessment for Worldwide Invasion and Spread of Tuta absoluta with a Focus on Sub-Saharan Africa: Implications for Phytosanitary Measures and Management.
To support management decisions, molecular characterization of data and geo-reference of incidence records of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) were combined with data on the biology and ecology of the pest to estimate its climatic suitability and potential spread at regional and global scale. A CLIMEX model was developed and used for the global prediction of current and future climate-induced changes in the distributional shifts of T. absoluta. Results revealed that temperature and moisture characterized T. absoluta population growth while the pest ability to survive the cold, hot, wet and dry stress conditions are the primary characteristics defining its range frontiers. Simulated irrigation also played an important role in the model optimization. Model predictions suggest that T. absoluta represents an important threat to Africa, Asia, Australia, Northern Europe, New Zealand, Russian Federation and the United States of America (USA). Under climate change context, future predictions on distribution of T. absoluta indicated that the invasive nature of this pest will result in significant crop losses in certain locations whereas some parts of Africa may witness diminution in ranges. The following scenarios may occur: 1) T. absoluta damage potential may upsurge moderately in areas of Africa where the pest currently exists; 2) a range diminution in temperate to Sahel region with moderate upsurge in damage potential; 3) a range expansion in tropical Africa with reasonable upsurge of damage potential. These possible outcomes could be explained by the fact that the continent is already warm, with the average temperature in majority of localities near the threshold temperatures for optimal development and survival of T. absoluta. Outputs from this study should be useful in helping decision-makers in their assessment of site-specific risks of invasion and spread of T. absoluta with a view to developing appropriate surveillance, phytosanitary measures and management strategies.


Scale-freeness or partial synchronization in neural mass phase oscillator networks: Pick one of two?
Modeling and interpreting (partial) synchronous neural activity can be a challenge. We illustrate this by deriving the phase dynamics of two seminal neural mass models: the Wilson-Cowan firing rate model and the voltage-based Freeman model. We established that the phase dynamics of these models differed qualitatively due to an attractive coupling in the first and a repulsive coupling in the latter. Using empirical structural connectivity matrices, we determined that the two dynamics cover the functional connectivity observed in resting state activity. We further searched for two pivotal dynamical features that have been reported in many experimental studies: (1) a partial phase synchrony with a possibility of a transition towards either a desynchronized or a (fully) synchronized state; (2) long-term autocorrelations indicative of a scale-free temporal dynamics of phase synchronization. Only the Freeman phase model exhibited scale-free behavior. Its repulsive coupling, however, let the individual phases disperse and did not allow for a transition into a synchronized state. The Wilson-Cowan phase model, by contrast, could switch into a (partially) synchronized state, but it did not generate long-term correlations although being located close to the onset of synchronization, i.e. in its critical regime. That is, the phase-reduced models can display one of the two dynamical features, but not both.


A factor V genetic component differing from factor V R506Q contributes to the activated protein C resistance phenotype.
Factor V gene polymorphisms were investigated to detect components that may contribute to the activated protein C (APC) resistance phenotype in patients with venous thromboembolism. A specific factor V gene haplotype (HR2) was defined by six polymorphisms and its frequency was found to be similar in normal subjects coming from Italy (0.08), India (0.1), and Somalia (0.08), indicating that it was originated by ancestral mutational events. The relationship between the distribution of normalized APC ratios obtained with the functional assay and haplotype frequency was analyzed in patients heterozygous for factor V R506Q (factor V Leiden). The HR2 haplotype was significantly more frequent in patients with ratios below the 15th percentile than in those with higher ratios or in normal controls. Moreover, the study of 10 patients with APC resistance in the absence of the factor V R506Q mutation showed a 50-fold higher frequency of HR2 homozygotes. The HR2 haplotype was associated with significantly lower APC ratios both in patients with venous thromboembolism and in age- and sex-matched controls. However, the two groups showed similar HR2 haplotype frequencies. Plasma mixing experiments showed that an artificially created double heterozygote for the factor V R506Q mutation and the HR2 haplotype had an APC ratio lower than that expected for a simple R506Q heterozygote. Time-course experiments evaluating the decay of factor V in plasma showed the normal stability of the molecule encoded by the factor V gene marked by the HR2 haplotype, which ruled out the presence of a pseudo-homozygous APC resistance mechanism. Our results provide new insights into the presence of factor V genetic components other than the factor V R506Q that are able to contribute to the APC resistance phenotype in patients with venous thromboembolism.


What Is a Rheumatologist and How Do We Make One?
Graduate medical education is a critical time in the training of a rheumatologist, and purposeful evaluation of abilities during this time is essential for long-term success as an independent practitioner. The internal medicine subspecialties collectively developed a uniform set of reporting milestones by which trainees can be assessed and receive formative feedback, providing clarity of accomplishment as well as areas for improvement in training. Furthermore, the reporting milestones provide a schema for assessment and evaluation of fellows by supervisors. The internal medicine subspecialties were also tasked with considering entrustable professional activities (EPAs), which define the abilities of a subspecialty physician who has attained sufficient mastery of the field to be accountable to stakeholders and participate in independent practice. Although EPAs have been established for a few specialties, they had not yet been described for rheumatology. EPAs have value as descriptors of the comprehensive abilities, knowledge, and skills of a practicing rheumatologist. The rheumatology EPAs have a role in defining a specialist in rheumatology upon completion of training, and also represent the ways our specialty defines our abilities that are enduring throughout practice. We describe the collaborative process of the development of both the subspecialty reporting milestones and the rheumatology EPAs. The reporting milestones evolved through discussions and collaborations among representatives from the Association of Specialty Professors, the Alliance for Academic Internal Medicine, the American Board of Internal Medicine, and the Accreditation Council for Graduate Medical Education. The EPAs were a product of deliberations by the Next Accreditation System (NAS) working group of the American College of Rheumatology (ACR) Committee on Rheumatology Training and Workforce Issues. Twenty-three subspecialty reporting milestones and 14 rheumatology EPAs were advanced and refined over the course of 3 subspecialty reporting milestone development summits and 3 ACR NAS working group meetings, respectively. The subspecialty reporting milestones and rheumatology EPAs presented here stipulate reasonable and measurable expectations for rheumatologists-in-training. Together, these tools aim to promote enrichment and greater accountability in the training of fellows. Additionally, the EPAs define, for all stakeholders, the expertise of a rheumatologist in practice.


Association of individual hnRNP proteins and snRNPs with nascent transcripts.
As they are transcribed, RNA polymerase II transcripts (hnRNAs or pre-mRNAs) associate with hnRNP proteins and snRNP particles, and the processing of pre-mRNA occurs within these ribonucleoprotein complexes. To better understand the relationship between hnRNP proteins and snRNP particles and their roles in mRNA formation, we have visualized them as they associate with nascent transcripts on the polytene chromosomes of Drosophila melanogaster salivary glands. Simultaneous pairwise detection of the abundant hnRNP proteins hrp36, hrp40, and hrp48 by direct double-label immunofluorescence microscopy reveals all of these proteins are bound to most transcripts, but their relative amounts on different transcripts are not fixed. Numerous differences in the relative amounts of snRNP particles and hnRNP proteins on nascent transcripts are also observed. These observations directly demonstrate that individual hnRNP proteins and snRNP particles are differentially associated with nascent transcripts and suggest that different pre-mRNAs bind different combinations of these factors to form transcript-specific, rather than a single type of, hnRNA-hnRNP-snRNP complexes. The distinct and specific constellation of hnRNP proteins and snRNP particles that assembles on different pre-mRNAs is likely to affect the fate and pathway of processing of these transcripts.


Protocol for the development of a core domain set for hidradenitis suppurativa trial outcomes.
Randomised controlled trials (RCTs) should have well-defined primary and secondary outcomes to answer questions generated by the main hypotheses. However, for the chronic, inflammatory skin disease hidradenitis suppurativa (HS), the reported outcome measures are numerous and diverse. A recent systematic review found a total of 30 outcome measure instruments in 12 RCTs. This use of a broad range of outcome measures can increase difficulties in interpretation and comparison of results and may potentially obstruct appropriate evidence synthesis by causing reporting bias. One strategy for dealing with these problems is to develop a core outcome set (COS). A COS is a list of outcomes that are meant as mandatory and should be measured and reported in all clinical trials. The aim of this study is to develop a COS for the management of HS. An international steering group of researchers, clinicians and a patient research partner will guide the COS development. 6 stakeholder groups are involved: patients, dermatologists, surgeons, nurses, industry representatives and drug regulatory authorities. A 1:1 ratio of patients:healthcare professionals is aimed for. The initial list of candidate items will be obtained by combining three data sets: (1) a systematic review of the literature, (2) US and Danish qualitative interview studies involving patients with HS and (3) an online healthcare professional (HCP) item generation survey. To reach consensus on the COS, 4 anonymous online Delphi rounds are then planned together with 2 face-to-face consensus meetings (1 in Europe and 1 in the USA) to ensure global representation. The study will be performed according to the Helsinki declaration. All results from the study, including inconclusive or negative results, will be published in peer-reviewed indexed journals. The study will involve different stakeholder groups to ensure that the developed COS will be suitable and well accepted.


Egg Albumin as a Protein Marker to Study Dispersal of Noctuidae in the Agroecosystem.
Knowledge of dispersal and spatial dynamics of pest populations is fundamental for implementation of integrated pest management and integrated resistance management. This study evaluated 1) the effectiveness of egg white albumin protein to mark larvae and adults of two polyphagous and highly mobile pests, Spodoptera frugiperda (J.E. Smith) (fall armyworm) and Helicoverpa zea (Boddie) (corn earworm) (Lepidoptera: Noctuidae), and 2) the sensitivity of polyvinylidene difluoride membrane (dot blot) in detecting albumin on marked insects. Laboratory and field experiments tested egg albumin as a protein marker, which was detected using two enzyme-linked immunosorbent assay (ELISA), microplate, and dot blot. In the laboratory, 100% of the moths sprayed with 20% egg white solution acquired the albumin marker, which was detected through the last time point tested (5 d) after application. Egg albumin was not effective at long-term marking of larvae, detected only prior the molting to the next instar. Albumin application in field cages resulted in a high percentage of moths detected as marked at 24 h and 5 d for both species. Egg albumin applied in the open field resulted in 15% of the recaptured corn earworm moths marked with most of them collected 150 m from the application area, although some were captured as far as 1,600 m within approximately 6 d after adult emergence. The results indicated egg albumin is a suitable marker to study the dispersion of fall armyworm and corn earworm in the agroecosystem and dot blot was as effective to detect egg albumin as was indirect ELISA.


Changes in consistency patterns of click frequency content over time of an echolocating Atlantic bottlenose dolphin.
Echolocation clicks were recorded from an Atlantic bottlenose dolphin Tursiops truncatus trained to discriminate frequency filtered phantom targets in 1998 and in 2004. These clicks showed consistency within their spectra intensity profiles but only in a certain band of frequencies. In 2004 almost all the clicks were consistent within the 0-42 kHz band regardless of the presented target or the click source level. This region corresponded with previous data showing that in 2004 the dolphin perceived frequencies only from within the 29-42 kHz band during echolocation. Above 42 kHz the consistency was lost. In 1998 the consistent region was found only in the 90-100 kHz band showing a shift had occurred with time. This suggests the dolphin's echolocation strategy for these discrimination tasks centered on the use of clicks with the same controlled standard frequency content in a certain frequency band to investigate different targets. This consistent region shifted over time to maintain maximum signal to noise ratio of the echoes given certain changing limitations to the echolocation system. The shift in consistency over time indicates these consistent regions were not simply artifacts of click production but rather an active control of frequency content.


Mutational hotspot in exon 20 of PIK3CA in breast cancer among Singapore Chinese.
The recent identification of somatic mutations in the catalytic region of PIK3 (PIK3CA) in breast cancer and demonstration of their oncogenic function has implicated PIK3CA in mammary carcinogenesis. To investigate possible ethnic differences in patterns of PIK3CA mutations in Singaporean Chinese breast cancer and to characterize these in a panel of cell lines, we sequenced exons 9 and 20 in 80 primary tumors, 19 breast cancer cell lines and 7 normal human mammary epithelial cells (HMECs). Searching for novel hotspots of mutation, we sequenced additional exons (1, 2, 6, 7, 14 and 18) in 20 primary tumors and 6 breast cancer cell lines. We detected 33 point mutations in 31 of 80 (39%) breast cancers, and 11 mutations in 10 of 19 (53%) breast cancer cell lines. No mutations were detected in normal breast tissue adjacent to the tumor, or in the 6 normal HMECs. The exon 20 A3140G (H1047R) substitution was identified most frequently (22/31, 71%) and showed a significant association with patient age (p = 0.043) and stage of the disease (p = 0.025), but not with ER/PR status or histological grade of the tumor. The incidence of point mutations in PIK3CA, the A3140G substitution in particular, in Singapore breast cancers are among the most frequent reported to date for any gene in breast cancer. The results suggest that mutation of PIK3CA might contribute to development of early stage breast cancer and could provide a potent target for early diagnosis and therapy.


Experimental model of proliferative vitreoretinopathy (PVR) in the vitrectomised eye: effect of silicone oil.
Silicone oil alone or as an adjunct to vitrectomy is widely used for the treatment of retinal detachment complicated by proliferative vitreoretinopathy (PVR). It appears to reduce the tendency for recurrent traction detachment, but the mechanism of this action is obscure. We have studied the effects of silicone oil in an experimental animal model. Twenty five thousand homologous fibroblasts were injected into the aphakic vitrectomised rabbit eye, and traction detachment of the vascularised retina (medullary rays) regularly resulted. Intraocular silicone oil did not influence the development of this experimental detachment.


Effect of endotoxin on colonisation of Campylobacter jejuni in infant mice.
An infant mouse model has been used to investigate the colonisation of the intestine by Campylobacter jejuni and the effect of endotoxin (Escherichia coli O26: B6) on the initial stage of this process. Endotoxin injected 1 or 16 h before the bacterial challenge had no effect on the growth of campylobacters but endotoxin injected 4 to 10 h before the bacterial challenge caused a bacteriostatic effect on the growth of campylobacters which lasted for one day. The bacteriostatic effect was evident both in the small intestine and in the distal part of the intestine containing caecum and colon. The mechanism of the bacteriostatic effect of endotoxin could not be explained in the study, but is thought to be non-immunological because it developed so rapidly. Oral and parenteral iron administered as ammonium ferric citrate or iron dextran, respectively, were used in an attempt to reverse the bacteriostatic effect. High oral doses of iron (0.5 mg per animal) were effective but small doses (0.5 mg per animal) were ineffective. Parenteral iron administration had a delayed effect on the reversal of the bacteriostatic effect of endotoxin. Transferrin administered orally caused a clear bacteriostatic effect in both endotoxin pretreated and untreated mice. Campylobacter counts were always lower in the small intestine than in the large intestine both in control and in endotoxin pretreated mice. This indicates that the large intestine is the primary ecological niche where campylobacters colonise mice.


Association of sleep and fatigue with decision regret among critical care nurses.
The effects of inadequate sleep on clinical decisions may be important for patients in critical care units, who are often more vulnerable than patients in other units. Fatigued nurses are more likely than well-rested nurses to make faulty decisions that lead to decision regret, a negative cognitive emotion that occurs when the actual outcome differs from the desired or expected outcome. To examine the association between selected sleep variables, impairment due to fatigue, and clinical-decision self-efficacy and regret among critical care nurses. Decision regret was the primary outcome variable. Methods A nonexperimental, descriptive design and extant measures were used to obtain data from a random sample of full-time nurses. Binary logistic regression models were used to examine the association between sleep variables, fatigue, and clinical-decision self-efficacy and regret. The discrimination of the models was compared with the C statistic, the area under the receiver operating characteristic curve. A total of 605 nurses returned the questionnaires (17% response rate). Among these, decision regret was reported by 157 of 546 (29%). Nurses with decision regret reported more fatigue, more daytime sleepiness, less intershift recovery, and worse sleep quality than did nurses without decision regret. Being male, working a 12-hour shift, and clinical-decision satisfaction were significantly associated with decision regret (C statistic, 0.719; SE, 0.024). Nurses who experience impairments due to fatigue, loss of sleep, and inability to recover between shifts are more likely than unimpaired nurses to report decision regret.


Whole body vibration and dynamic restraint.
The purpose of this study was to identify changes due to whole body vibration in peroneus longus (PL) activation following ankle inversion perturbation. Participants were 22 (age 22.1 +/- 1.8 yrs, ht 168.8 +/- 8.2 cm, mass 65.5 +/- 11.2 kg) physically active male and female students with no recent history of lower extremity injury. Measurements of PL electromechanical delay (EMD), reaction time, and muscle activation were collected from two groups (WBV and control) over 3 time intervals (pretreatment, posttreatment, and 30 min posttreatment). Two-way ANOVAs were used to compare groups over time for all dependent variables. No group x time interactions were detected (p < 0.05) for any of the dependent variables. Whole body vibration did not alter PL EMD, reaction time, peak EMG, or average EMG. The use of WBV for enhancing ankle dynamic stability was not supported by this study. However, more data are needed to determine if WBV is an effective intervention in other areas of injury prevention or rehabilitation. These data were not consistent with the hypothesis that WBV enhances muscle spindle sensitivity.


Characteristics of Copayment Offsets for Prescription Drugs in the United States.
Despite ongoing debate regarding the high prices that patients pay for prescription drugs, to our knowledge, little is known regarding the use of coupons, vouchers, and other types of copayment "offsets" that reduce patients' out-of-pocket drug spending. Although offsets reduce patients' immediate cost burden, they may encourage the use of higher-cost products and diminish health insurers' ability to optimize pharmaceutical value. To examine the drugs most commonly covered by offsets, the percentage of out-of-pocket costs covered by offsets, and the characteristics of patients using offsets for retail pharmacy transactions in the United States in 2017 through 2019. A retrospective cohort analysis was conducted of a 5% nationally random sample of anonymized pharmacy claims from IQVIA's Formulary Impact Analyzer, which captures more than 60% of all US pharmacy transactions. This analysis focused on 631 249 individuals who used at least 1 offset between October 1, 2017, and September 30, 2019. Offset source, types of drugs covered by offsets, offset dollar value and percentage of out-of-pocket payment covered, and county characteristics of offset recipients. The 631 249 individuals in the study (361 855 female participants [57.3%]; mean [SD] age, 45.7 [18.6] years) had approximately 33 million prescription fills, of which 12.8% had an offset used. Of these, 50.2% originated from a pharmaceutical manufacturer, 47.2% originated from a pharmacy or pharmacy benefit manager (PBM), and 2.6% originated from a state assistance program. A total of 80.0% of manufacturer-sponsored offsets were concentrated among 6.2% of unique products, and 79.9% of pharmacy-PBM offsets were concentrated among 4.9% of unique products. Most manufacturer offsets (88.2%) were for branded products, while most pharmacy-PBM offsets were for generic products (90.5%). The median manufacturer offset was $51.00, covering 87.1% of out-of-pocket costs; the median pharmacy-PBM offset was $16.30, covering 39.3% of out-of-pocket costs. There was no meaningful association between offset magnitude and county-level income, health insurance coverage, or race/ethnicity. In this analysis of patient-level pharmacy claims from 2017 to 2019, approximately half of all offsets involved pharmacy-PBM contractual arrangements, and half were offered by manufacturers. All offsets were associated with a significant reduction in patients' out-of-pocket costs, were highly concentrated among a few drugs, and were generally not more generous among individuals in counties with lower income or larger Black or uninsured populations.


Self-sampling for oropharyngeal and rectal specimens to screen for sexually transmitted infections: acceptability among men who have sex with men.
To explore the feasibility and acceptability of self-sampling for oropharyngeal and rectal specimens to screen for sexually transmitted infections (STIs) among men who have sex with men (MSM). Participant's willingness to self-sample at home was also explored. Participants of a study to evaluate the sensitivity and specificity of self versus nurse taken oropharyngeal and rectal specimens were surveyed to assess the feasibility and acceptability of self-sampling using specimen collection methods (gargle, OraSure mouth pad to collect oropharyngeal specimens and APTIMA unisex swabs to collect rectal and pharyngeal specimens). Acceptability was measured using a five-point Likert-type response scale (for example, 1 = strongly disagree; 5 = strongly agree). Open-ended questions explored participants' experiences of self-sampling. Of 334 eligible MSM, 301 (90%) participated in the study. Altogether, 301 participants self-sampled using gargle and rectal and pharyngeal swabs and 288 using mouth pad. Complete questionnaire data from 274 participants showed that feasibility and acceptability of self-sampling using gargle and mouth pad was higher (92%) than pharyngeal swabs (76%). Rectal swabs were acceptable to 82% participants. Despite some discomfort and difficulty in using swabs, 76% were willing to use all four methods for self-sampling in the future. Home sampling was acceptable (84%) as it was perceived to be less intrusive and more convenient than a clinic visit and likely to reduce genitourinary medicine (GUM) waiting time. Self-sampling for rectal and oropharyngeal specimens is feasible and acceptable to MSM. Self-sampling can be offered as an alternative to clinic-based testing and has the potential to improve choice, access and uptake of screening for STIs.


Effects of multiple monthly doses of ivermectin on adult Onchocerca volvulus.
This paper assesses the effects on adult Onchocerca volvulus of monthly doses of ivermectin (150 micrograms/kg) given over 4, 8, and 12 months to patients in Guatemala. Nodules were removed 4 months after the last dose; the adult O. volvulus were extracted by collagenase digestion, studied by histological techniques, and compared with worms from untreated patients. Twelve monthly doses killed a proportion of the adult worms (12% of males and 22% of females), leaving the remainder relatively unaffected and the females slowly resuming embryogenesis. After 8 and 12 doses, a number of female worms had resumed embryogenesis in 1 genital tract only, and in 1 female a total degeneration of 1 ovary was seen. Ivermectin also led to a marked drop in the number of male worms in nodules. No serious adverse reactions occurred and the treatment was well accepted.


Comparing urine samples and cervical swabs for Chlamydia testing in a female population by means of Strand Displacement Assay (SDA).
There has been an increasing number of diagnosed cases of Chlamydia trachomatis in many countries, in particular among young people. The present study was based on a growing request to examine urine as a supplementary or primary specimen in screening for Chlamydia trachomatis in women, with the Becton Dickinson ProbeTec (BDPT) Strand Displacement Assay (SDA). Urine samples may be particularly important in screening young people who are asymptomatic. A total of 603 women aged 15 and older were enrolled from the Sexually Transmitted Infection (STI) clinic at Haukeland University Hospital, Norway, in 2007. Only 31 women were older than 35 years. Cervical swabs and urine samples were tested with BDPT for all participants. In cases of discrepant test results from a given patient, both samples were retested by Cobas TaqManCT and a Polymerase Chain Reaction (PCR)-method (in-house). Prevalence of C. trachomatis, sensitivity, and specificity were estimated by latent class analysis using all test results available. Bootstrap BC confidence intervals (10,000 computations) were estimated for sensitivity and specificity, and their differences in cervix vs. urine tests. A total of 1809 specimens were collected from 603 patients. 80 women (13.4%) were positive for C. trachomatis. Among these, BDPT identified 72 and 73 as positive in cervix and urine samples, respectively. Of the 523 C. trachomatis negative women, BDPT identified 519 as negative based on cervical swabs, and 514 based on urine samples. Sensitivity for cervical swabs and urine samples with the BDPT were 89.0% (95% CI 78.8, 98.6) and 90.2% (95% CI 78.1, 95.5), respectively. The corresponding values for specificity were 99.2% (95% CI 98.3, 100) and 98.3% (95% CI 96.4, 100). This study indicates that urine specimens are adequate for screening high-risk groups for C. trachomatis by the SDA method (BDPT). Such an approach may facilitate early detection and treatment of the target groups for screening, and be cost-effective for patients and the health services.


Long-term evolution of patient-reported outcome measures in spinocerebellar ataxias.
To study the long-term evolution of patient-reported outcome measures (PROMs) in the most common spinocerebellar ataxias (SCAs), we analyzed 8 years follow-up data of the EUROSCA Natural History Study, a cohort study of 526 patients with SCA1, SCA2, SCA3 and SCA6. To assess the functional capacity in daily living, we used the functional assessment (part IV) of the Unified Huntington's Disease Rating Scale (UHDRS-IV), for health-related quality of life the visual analogue scale of the EuroQol Five Dimensions Questionnaire (EQ-5D VAS), and for depressive symptoms the Patient Health Questionnaire (PHQ-9). Severity of ataxia was assessed using the Scale for the Assessment and Rating of Ataxia (SARA) and neurological symptoms other than ataxia with the Inventory of Non-Ataxia Signs (INAS). UHDRS-IV [SCA1: - 1.35 (0.12); SCA2: - 1.15 (0.11); SCA3: - 1.16 (0.11); SCA6: - 0.99 (0.12)] and EQ-5D [SCA1: - 2.88 (0.72); SCA2: - 1.97 (0.49); SCA3: - 2.06 (0.55); SCA6: - 1.03 (0.57)] decreased linearly, whereas PHQ-9 increased [SCA1: 0.15 (0.04); SCA2: 0.09 (0.03); SCA3: 0.06 (0.04); SCA6: 0.07 (0.04)] during the observational period. Standard response means (SRMs) of UHDRS-IV (0.473-0.707) and EQ-5D VAS (0.053-0.184) were lower than that of SARA (0.404-0.979). In SCA1, higher SARA scores [- 0.0288 (0.01), p = 0.0251], longer repeat expansions [- 0.0622 (0.02), p = 0.0002] and the presence of cognitive impairment at baseline [- 0.5381 (0.25), p = 0.0365] were associated with faster UHDRS-IV decline. In SCA3, higher INAS counts were associated with a faster UHDRS-IV decline [- 0.05 (0.02), p = 0.0212]. In SCA1, PHQ-9 progression was faster in patients with cognitive impairment [0.14 (0.07); p = 0.0396]. In the common SCAs, PROMs give complementary information to the information provided by neurological scales. This underlines the importance of PROMs as additional outcome measures in future interventional trials.


Mailed fecal immunochemical test outreach for colorectal cancer screening: Summary of a Centers for Disease Control and Prevention-sponsored Summit.
Uptake of colorectal cancer screening remains suboptimal. Mailed fecal immunochemical testing (FIT) offers promise for increasing screening rates, but optimal strategies for implementation have not been well synthesized. In June 2019, the Centers for Disease Control and Prevention convened a meeting of subject matter experts and stakeholders to answer key questions regarding mailed FIT implementation in the United States. Points of agreement included: 1) primers, such as texts, telephone calls, and printed mailings before mailed FIT, appear to contribute to effectiveness; 2) invitation letters should be brief and easy to read, and the signatory should be tailored based on setting; 3) instructions for FIT completion should be simple and address challenges that may lead to failed laboratory processing, such as notation of collection date; 4) reminders delivered to initial noncompleters should be used to increase the FIT return rate; 5) data infrastructure should identify eligible patients and track each step in the outreach process, from primer delivery through abnormal FIT follow-up; 6) protocols and procedures such as navigation should be in place to promote colonoscopy after abnormal FIT; 7) a high-quality, 1-sample FIT should be used; 8) sustainability requires a program champion and organizational support for the work, including sufficient funding and external policies (such as quality reporting requirements) to drive commitment to program investment; and 9) the cost effectiveness of mailed FIT has been established. Participants concluded that mailed FIT is an effective and efficient strategy with great potential for increasing colorectal cancer screening in diverse health care settings if more widely implemented.


Global, Regional, National, and Subnational Big Data to Inform Health Equity Research: Perspectives from the Global Burden of Disease Study 2017.
Everyone deserves a long and healthy life, but in reality, health outcomes differ across populations. We use results from the Global Burden of Disease Study 2017 (GBD 2017) to report patterns in the burden of diseases, injuries, and risks at the global, regional, national, and subnational level, and by sociodemographic index (SDI), from 1990 to 2017. GBD 2017 undertook a systematic analysis of published studies and available data providing information on prevalence, incidence, remission, and excess mortality. We computed prevalence, incidence, mortality, life expectancy, healthy life expectancy, years of life lost due to premature mortality, years lived with disability, and disability-adjusted life years with 95% uncertainty intervals for 23 age groups, both sexes, and 918 locations, including 195 countries and territories and subnational locations for 16 countries from 1990 to 2017. We also computed SDI, a summary indicator combining measures of income, education, and fertility. There were wide disparities in the burden of disease by SDI, with smaller burdens in affluent countries and in specific regions within countries. Select diseases and risks, such as drug use disorders, high blood pressure, high body mass index, diet, high fasting plasma glucose, smoking, and alcohol use disorders warrant increased global attention and indicate a need for greater investment in prevention and treatment across the life course. Policymakers need a comprehensive picture of what risks and causes result in disability and death. The GBD provides the means to quantify health loss: these findings can be used to examine root causes of disparities and develop programs to improve health and health equity.


Human cochlear tuning estimates from stimulus-frequency otoacoustic emissions.
Two objective measures of human cochlear tuning, using stimulus-frequency otoacoustic emissions (SFOAE), have been proposed. One measure used SFOAE phase-gradient delay and the other two-tone suppression (2TS) tuning curves. Here, it is hypothesized that the two measures lead to different frequency functions in the same listener. Two experiments were conducted in ten young adult normal-hearing listeners in three frequency bands (1-2 kHz, 3-4 kHz and 5-6 kHz). Experiment 1 recorded SFOAE latency as a function of stimulus frequency, and experiment 2 recorded 2TS iso-input tuning curves. In both cases, the output was converted into a sharpness-of-tuning factor based on the equivalent rectangular bandwidth. In both experiments, sharpness-of-tuning curves were shown to be frequency dependent, yielding sharper relative tuning with increasing frequency. Only a weak frequency dependence of the sharpness-of-tuning curves was observed for experiment 2, consistent with objective and behavioural estimates from the literature. Most importantly, the absolute difference between the two tuning estimates was very large and statistically significant. It is argued that the 2TS estimates of cochlear tuning likely represents the underlying properties of the suppression mechanism, and not necessarily cochlear tuning. Thus the phase-gradient delay estimate is the most likely one to reflect cochlear tuning.


Single-cell eQTL mapping in yeast reveals a tradeoff between growth and reproduction.
Expression quantitative trait loci (eQTLs) provide a key bridge between noncoding DNA sequence variants and organismal traits. The effects of eQTLs can differ among tissues, cell types, and cellular states, but these differences are obscured by gene expression measurements in bulk populations. We developed a one-pot approach to map eQTLs in Saccharomyces cerevisiae by single-cell RNA sequencing (scRNA-seq) and applied it to over 100,000 single cells from three crosses. We used scRNA-seq data to genotype each cell, measure gene expression, and classify the cells by cell-cycle stage. We mapped thousands of local and distant eQTLs and identified interactions between eQTL effects and cell-cycle stages. We took advantage of single-cell expression information to identify hundreds of genes with allele-specific effects on expression noise. We used cell-cycle stage classification to map 20 loci that influence cell-cycle progression. One of these loci influenced the expression of genes involved in the mating response. We showed that the effects of this locus arise from a common variant (W82R) in the gene GPA1 , which encodes a signaling protein that negatively regulates the mating pathway. The 82R allele increases mating efficiency at the cost of slower cell-cycle progression and is associated with a higher rate of outcrossing in nature. Our results provide a more granular picture of the effects of genetic variants on gene expression and downstream traits.


Longitudinal Faculty Development Program to Promote Effective Observation and Feedback Skills in Direct Clinical Observation.
We developed a longitudinal faculty development program to maximize faculty training in direct clinical observation (DCO) and feedback, as there was a perceived need for higher quality of DCO and feedback. To achieve this, we created a behaviorally anchored DCO instrument and worked to improve faculty skills in this area. We describe an innovative model of faculty training that is learner centered and reinforces evidence-based principles of effective feedback that are introduced and then repeated in all sessions. The training centers on both peer-led observation of and feedback on faculty learners' recorded DCO feedback encounters, and is guided by our DCO instrument. Residents and faculty completed surveys to assess program impact. Qualitative responses were analyzed for themes. The Wilcoxon signed rank test was used to examine significance of difference in feedback quality before and after DCO faculty development education sessions. Our faculty development program has been well received and had a significant impact on quality of faculty feedback, as rated by resident learners. Our faculty development model is effective at growing faculty learners' DCO and feedback skills. Potential strengths of this program include the use of a behaviorally anchored DCO instrument, longitudinal and experiential faculty development, and use of small-group peer review of recorded faculty feedback encounters. We have found that when their learning needs are attended to, faculty learners cultivate a deep appreciation for principles of effective feedback. In fact, faculty feedback skills can be enhanced in the eyes of resident learners.


Patterns of dolphin bycatch in a north-western Australian trawl fishery.
The bycatch of small cetaceans in commercial fisheries is a global wildlife management problem. We used data from skippers' logbooks and independent observers to assess common bottlenose dolphin (Tursiops truncatus) bycatch patterns between 2003 and 2009 in the Pilbara Trawl Fishery, Western Australia. Both datasets indicated that dolphins were caught in all fishery areas, across all depths and throughout the year. Over the entire datasets, observer reported bycatch rates (n = 52 dolphins in 4,124 trawls, or 12.6 dolphins/1,000 trawls) were ca. double those reported by skippers (n = 180 dolphins in 27,904 trawls, or 6.5 dolphins/1,000 trawls). Generalised Linear Models based on observer data, which better explained the variation in dolphin bycatch, indicated that the most significant predictors of dolphin catch were: (1) vessel--one trawl vessel caught significantly more dolphins than three others assessed; (2) time of day--the lowest dolphin bycatch rates were between 00:00 and 05:59; and (3) whether nets included bycatch reduction devices (BRDs)--the rate was reduced by ca. 45%, from 18.8 to 10.3 dolphins/1,000 trawls, after their introduction. These results indicated that differences among vessels (or skippers' trawling techniques) and dolphin behavior (a diurnal pattern) influenced the rates of dolphin capture; and that spatial or seasonal adjustments to trawling effort would be unlikely to significantly reduce dolphin bycatch. Recent skipper's logbook data show that dolphin bycatch rates have not declined since those reported in 2006, when BRDs were introduced across the fishery. Modified BRDs, with top-opening escape hatches from which dolphins might escape to the surface, may be a more effective means of further reducing dolphin bycatch. The vulnerability of this dolphin population to trawling-related mortality cannot be assessed in the absence of an ongoing observer program and without information on trawler-associated dolphin community size, broader dolphin population size and connectivity with adjacent populations.


Fecal microbiota transplantation as a new therapy: from Clostridioides difficile infection to inflammatory bowel disease, irritable bowel syndrome, and colon cancer.
Fecal microbiota transplantation (FMT) represents the most effective means of therapeutically manipulating the gastrointestinal microbiome. Originally employed as a treatment of last-resort in patients with life-threatening Clostridioides difficile infection (CDI), FMT gained widespread acceptance during the CDI epidemic, where it achieved resolution rates approaching 100%. Following our newfound appreciation for the role of the gut microbiome in both health and disease and owing to FMT's unique mechanism/s of action, FMT is rapidly advancing as an effective treatment for a number of conditions in which the gastrointestinal microbiome is thought to play a role. We review the role of FMT from its beginnings in CDI to its expansion into inflammatory bowel disease, irritable bowel syndrome, and colon cancer.


Frequency of symptoms associated with gallstone disease: a hospital-based cross sectional study.
This study investigated the likely outcomes of gallstone disease and frequency of symptoms associated with this. This cross sectional study was performed at the Aliebn Abitaleb hospital in Rafsanjan over a period of twelve months from 2000 to 2001. A total sample of 70 individuals were included in the study, interviewing them using of a questionnaire covering data in terms of demographic features and medical history.The majority of patients belonged to the age group of 41 to 60 years (44.3% of all patients), while the lowest frequency was in the age group of 20 years or less. Frequency of females was higher than males. The frequency of female patients increased with the number of child birth, the highest being in women who had more than 5 child births. The most common complaint of pain was revealed as upper right abdominal quadrant (RUQ), which accounted for 95.7% of cases. Vomiting was observed in 52.9% of patients. In addition, the frequency of patients with chronic pain initiation was more than those with acute pain initiation. Among women, 17.5% had oral contraceptive pill constipation. It is worth noting that ultrasonography was the best diagnostic tool for evaluating patients. It is noteworthy that the most common symptom was right upper quadrant tenderness. Our data indicate that diabetes was the most common illness associated with gallstone disease and that 8.6% of patients had a family history of gallstone disease.


International matches elicit stable mechanical workload in high-level female ice hockey.
This study aimed to quantify in- and between-match characteristics and mechanical workload variations elicited by a congested schedule in high-level female ice hockey. Six players were monitored during four international pre-season exhibition matches against the same opponent. Two different methods (Player Load and Accel'Rate) were used to assess specific mechanical workload. Number of shifts and effective playing time per shift were significantly higher for period 2 (p = 0.03 for both). Mechanical workload intensity (i.e., relative and peak workload) showed a significant (p ≤ 0.05) decrease from period 1 to period 2 and period 3 (moderate-to-large Cohen's d). All workload variables remained stable between matches (p > 0.25). Team variability showed good-to-moderate CVs (< 10%) for all variables for in- and between-match variability. Accumulated workload computed with the Player Load method was threefold higher compared to the Accel'Rate method (+ 87.8% mean difference; large Cohen's d). These findings demonstrate that high-level female ice hockey-specific mechanical workload declines with reduced high-intensity output across periods, while it remains stable between matches against standardized opposition. This study strongly suggests that the present workload metrics could be used to determine the mechanical demand elicited by matches played against various opponents in real game conditions.


Generation of sub-10-fs pulses from a Kerr-lens mode-locked Cr(3+):LiCAF laser oscillator by use of third-order dispersion-compensating double-chirped mirrors.
Pulses as short as 9 fs at 220-mW average power and a 97-MHz repetition rate are generated from a cw Ti:sapphire-pumped Kerr-lens mode-locked Cr(3+)LiCAF laser oscillator employing broadband double-chirped mirrors for second- and third-order dispersion compensation. Fine adjustment of dispersion is accomplished with a fused-silica prism pair. The result demonstrates that Raman-induced self-frequency shifting of the pulse does not limit sub-10-fs pulse generation from colquiriite crystals.


Rate coefficients and reaction mechanism for the reaction of OH radicals with (E)-CF3CH═CHF, (Z)-CF3CH═CHF, (E)-CF3CF═CHF, and (Z)-CF3CF═CHF between 200 and 400 K: hybrid density functional theory and canonical variational transition state theory calculations.
The rate coefficients of ((E)-CF(3)CH═CHF, (Z)-CF(3)CH═CHF, (E)-CF(3)CF═CHF, and (Z)-CF(3)CF═CHF) + OH reactions were computed using M06-2X/6-31+G(d,p) theory in the temperature range of 200 and 400 K. The possible reaction mechanisms of the ((E)-CF(3)CH═CHF, (Z)-CF(3)CH═CHF, (E)-CF(3)CF═CHF, and (Z)-CF(3)CF═CHF) + OH reactions were examined. The rate coefficients for the addition and abstraction reactions were calculated using canonical variational transition state theory (CVT) and conventional transition state theory (CTST), respectively, and we concluded that abstraction reactions are negligible within the temperature range and addition reactions take the lead role. The small curvature tunnelling (SCT) was included in the computation of the rate coefficients. The temperature dependent rate expressions (in cm(3) molecule(-1) s(-1)) of the (E)-CF(3)CH═CHF, (Z)-CF(3)CH═CHF, (E)-CF(3)CF═CHF, and (Z)-CF(3)CF═CHF + OH reactions between 200 and 400 K are presented. The atmospheric lifetimes and global warming potentials (GWPs) of the test molecules were computed using the rate coefficients obtained in this study, and it is concluded that these molecules are very short-lived in the Earth's atmosphere with low GWPs.


Productivity Costs Associated With Breast Cancer Among Survivors Aged 18-44 Years.
No study has quantified productivity losses associated with breast cancer in younger women aged 18-44 years. This study estimated productivity costs, including work and home productivity losses, among younger women who reported ever receiving a breast cancer diagnosis. A two-part regression model and 2000-2010 National Health Interview Survey data were used to estimate the number of work and home productivity days missed because of breast cancer, adjusted for socioeconomic characteristics and comorbidities. Estimates for younger women were compared with those for women aged 45-64 years. Data were analyzed in 2013-2014. Per capita, younger women with breast cancer had annual losses of $2,293 (95% CI=$1,069, $3,518) from missed work and $442 (95% CI=$161, $723) from missed home productivity. Total annual breast cancer-associated productivity costs for younger women were $344 million (95% CI=$154 million, $535 million). Older women with breast cancer had lower per capita work loss productivity costs of $1,407 (95% CI=$899, $1,915) but higher total work loss productivity costs estimated at $1,072 million (95% CI=$685 million, $1,460 million) than younger women. Younger women with a history of breast cancer face a disproportionate share of work and home productivity losses. Although older women have lower per capita costs, total productivity costs were higher for older women because the number of older women with breast cancer is higher. The results underscore the importance of continued efforts by the public health community to promote and support the unique needs of younger breast cancer survivors.


Effects of dibutyryl cAMP on stanniocalcin and stanniocalcin-related protein mRNA expression in neuroblastoma cells.
Stanniocalcin is a polypeptide hormone that was first reported in fish as a regulator of mineral metabolism. Its recent identification in mammals has opened a new area of investigation in basic and clinical endocrinology. In the present study, regulation of the stanniocalcin (STC) and stanniocalcin related protein (STCrP) genes were investigated in mouse neuroblastoma cells (Neuro-2A) in relation to neuronal cell differentiation. Neuro-2A is an undifferentiated cell line that contains measurable levels of STCrP mRNA, but undetectable levels of STC mRNA. Treatment of the cells with either dbcAMP (1-4 mM) or 50 microM euxanthone (PW1) resulted in extensive differentiation and neurite outgrowth. However, only neurites of dbcAMP-treated cells developed varicosities, a phenotypic marker of axon formation. Furthermore, following differentiation induced by dbcAMP, there was an upregulation of STC and downregulation of STCrP mRNA levels. In the first 24 and 48 h of treatments, there was a maximum twofold induction and 1.5-fold reduction in STC and STCrP mRNAs respectively. Following 96 h of treatment, an additional 14-fold STC induction and 1.2-fold STCrP reduction were observed. The increase in STC mRNA levels was accompanied by a concomitant increase in axon-specific low molecular form microtubule-associated protein (MAP-2c) mRNA and varicosities on the neurites, suggesting a possible role for STC in axonogenesis. There was no induction of STC mRNA levels when PW1 was added into the culture media, whereas ionomycin (1-10 microM) had no observable effects on cell differentiation or STC/STCrP mRNA. Immunocytochemical staining of dbcAMP-treated cells revealed abundant levels of immunoreactive STC, particularly in the varicosities, with only weak staining in control, untreated cells. Antisense oligodeoxynucleotides transfection studies indicated that the expression of STC was a cause of varicosity formation and a consequence of cell differentiation. Our findings lend further support to the notion that STC is involved in the process of neural differentiation.


Highly anisotropic decay rates of single quantum dots in photonic crystal membranes.
We have measured the variation of the spontaneous emission rate with polarization for self-assembled single quantum dots in two-dimensional photonic crystal membranes. We observe a maximum anisotropy factor of 6 between the decay rates of the two bright exciton states. This large anisotropy is attributed to the substantially different projected local density of optical states for differently oriented dipoles in the photonic crystal.


Pneumococcal galactose catabolism is controlled by multiple regulators acting on pyruvate formate lyase.
Catabolism of galactose by Streptococcus pneumoniae alters the microbe's metabolism from homolactic to mixed acid fermentation, and this shift is linked to the microbe's virulence. However, the genetic basis of this switch is unknown. Pyruvate formate lyase (PFL) is a crucial enzyme for mixed acid fermentation. Functional PFL requires the activities of two enzymes: pyruvate formate lyase activating enzyme (coded by pflA) and pyruvate formate lyase (coded by pflB). To understand the genetic basis of mixed acid fermentation, transcriptional regulation of pflA and pflB was studied. By microarray analysis of ΔpflB, differential regulation of several transcriptional regulators were identified, and CcpA, and GlnR's role in active PFL synthesis was studied in detail as these regulators directly interact with the putative promoters of both pflA and pflB, their mutation attenuated pneumococcal growth, and their expression was induced on host-derived sugars, indicating that these regulators have a role in sugar metabolism, and multiple regulators are involved in active PFL synthesis. We also found that the influence of each regulator on pflA and pflB expression was distinct in terms of activation and repression, and environmental condition. These results show that active PFL synthesis is finely tuned, and feed-back inhibition and activation are involved.


In vivo and in vitro propagation and transmission of Toggenburg orbivirus.
The Toggenburg orbivirus (TOV), a recently discovered virus related to bluetongue virus (BTV), has been identified in goats in Switzerland, Italy and Germany. Isolation of TOV in vitro has not yet been achieved and the transmission mechanisms are still unknown. In the experimental infection of pregnant goats described here, TOV could not be detected in secretion/excretion samples or fetal blood. Material from the goat experiment was used as inoculum for propagating the virus in vitro. To enhance the infectivity of TOV several modified protocols, e.g. pretreatment of the virus with trypsin, polyethylene glycol-mediated infection and lipofection were applied. Isolation of TOV, attempts to infect Culicoides nubeculosus by feeding TOV-positive blood and intracerebral inoculation of newborn mice were unsuccessful. The results of these studies suggest that TOV requires specific but different factors than other BTVs for infection and replication outside of its natural caprine host.


Semaphorin 3A contributes to distal pulmonary epithelial cell differentiation and lung morphogenesis.
Semaphorin 3A (Sema3A) is a neural guidance cue that also mediates cell migration, proliferation and apoptosis, and inhibits branching morphogenesis. Because we have shown that genetic deletion of neuropilin-1, which encodes an obligatory Sema3A co-receptor, influences airspace remodeling in the smoke-exposed adult lung, we sought to determine whether genetic deletion of Sema3A altered distal lung structure. To determine whether loss of Sema3A signaling influenced distal lung morphology, we compared pulmonary histology, distal epithelial cell morphology and maturation, and the balance between lung cell proliferation and death, in lungs from mice with a targeted genetic deletion of Sema3A (Sema3A(-/-)) and wild-type (Sema3A(+/+)) littermate controls. Genetic deletion of Sema3A resulted in significant perinatal lethality. At E17.5, lungs from Sema3A(-/-) mice had thickened septae and reduced airspace size. Distal lung epithelial cells had increased intracellular glycogen pools and small multivesicular and lamellar bodies with atypical ultrastructure, as well as reduced expression of type I alveolar epithelial cell markers. Alveolarization was markedly attenuated in lungs from the rare Sema3A(-/-) mice that survived the immediate perinatal period. Furthermore, Sema3A deletion was linked with enhanced postnatal alveolar septal cell death. These data suggest that Sema3A modulates distal pulmonary epithelial cell development and alveolar septation. Defining how Sema3A influences structural plasticity of the developing lung is a critical first step for determining if this pathway can be exploited to develop innovative strategies for repair after acute or chronic lung injury.


A compendium of human genes regulating feeding behavior and body weight, its functional characterization and identification of GWAS genes involved in brain-specific PPI network.
Obesity is heritable. It predisposes to many diseases. The objectives of this study were to create a compendium of genes relevant to feeding behavior (FB) and/or body weight (BW) regulation; to construct and to analyze networks formed by associations between genes/proteins; and to identify the most significant genes, biological processes/pathways, and tissues/organs involved in BW regulation. The compendium of genes controlling FB or BW includes 578 human genes. Candidate genes were identified from various sources, including previously published original research and review articles, GWAS meta-analyses, and OMIM (Online Mendelian Inheritance in Man). All genes were ranked according to knowledge about their biological role in body weight regulation and classified according to expression patterns or functional characteristics. Substantial and overrepresented numbers of genes from the compendium encoded cell surface receptors, signaling molecules (hormones, neuropeptides, cytokines), transcription factors, signal transduction proteins, cilium and BBSome components, and lipid binding proteins or were present in the brain-specific list of tissue-enriched genes identified with TSEA tool. We identified 27 pathways from KEGG, REACTOME and BIOCARTA whose genes were overrepresented in the compendium. Networks formed by physical interactions or homological relationships between proteins or interactions between proteins involved in biochemical/signaling pathways were reconstructed and analyzed. Subnetworks and clusters identified by the MCODE tool included genes/proteins associated with cilium morphogenesis, signal transduction proteins (particularly, G protein-coupled receptors, kinases or proteins involved in response to insulin stimulus) and transcription regulation (particularly nuclear receptors). We ranked GWAS genes according to the number of neighbors in three networks and revealed 22 GWAS genes involved in the brain-specific PPI network. On the base of the most reliable PPIs functioning in the brain tissue, new regulatory schemes interpreting relevance to BW regulation are proposed for three GWAS genes (ETV5, LRP1B, and NDUFS3). A compendium comprising 578 human genes controlling FB or BW was designed, and the most significant functional groups of genes, biological processes/pathways, and tissues/organs involved in BW regulation were revealed. We ranked genes from the GWAS meta-analysis set according to the number and quality of associations in the networks and then according to their involvement in the brain-specific PPI network and proposed new regulatory schemes involving three GWAS genes (ETV5, LRP1B, and NDUFS3) in BW regulation. The compendium is expected to be useful for pathology risk estimation and for design of new pharmacological approaches in the treatment of human obesity.


Sub-Poissonian-light generation by postselection from twin beams.
States with sub-Poissonian photon-number statistics obtained by post-selection from twin beams are experimentally generated. States with Fano factors down to 0.62 and mean photon numbers around 12 are reached. Their quasi-distributions of integrated intensities attaining negative values are reconstructed. An intensified CCD camera with a quantum detection efficiency exceeding 20% is utilized both for post-selection and beam characterization. Experimental results are compared with theory that provides the optimum experimental conditions.


Drug-drug interaction (DDI) assessments of ruxolitinib, a dual substrate of CYP3A4 and CYP2C9, using a verified physiologically based pharmacokinetic (PBPK) model to support regulatory submissions.
Ruxolitinib is mainly metabolized by cytochrome P450 (CYP) enzymes CYP3A4 and CYP2C9 followed by minor contributions of other hepatic CYP enzymes in vitro. A physiologically based pharmacokinetic (PBPK) model was established to evaluate the changes in the ruxolitinib systemic exposures with co-administration of CYP3A4 and CYP2C9 perpetrators. The fractions metabolized in the liver via oxidation by CYP enzymes (fm,CYP3A4 = 0.75, fm,CYP2C9 = 0.19, and fm,CYPothers = 0.06) for an initial ruxolitinib model based on in vitro data were optimized (0.43, 0.56, and 0.01, respectively) using the observed exposure changes of ruxolitinib (10 mg) with co-administered ketoconazole (200 mg). The reduced amount of fm,CYP3A4 was distributed to fm,CYP2C9. For the initial ruxolitinib model with co-administration of ketoconazole, the area under the curve (AUC) increase of 2.60-fold was over-estimated compared with the respective observation (1.91-fold). With the optimized fm values, the predicted AUC ratio was 1.82. The estimated AUC ratios of ruxolitinib by co-administration of the moderate CYP3A4 inhibitor erythromycin (500 mg) and the strong CYP3A4 inducer rifampicin (600 mg) were within a 20% error compared with the clinically observed values. The PBPK modeling results may provide information on the labeling, i.e. supporting a dose reduction by half for co-administration of strong CYP3A4 inhibitors. Furthermore, an AUC increase of ruxolitinib in the absence or presence of the dual CYP3A4 and CYP2C9 inhibitor fluconazole (100-400 mg) was prospectively estimated to be 1.94- to 4.31-fold. Fluconazole simulation results were used as a basis for ruxolitinib dose adjustment when co-administering perpetrator drugs. A ruxolitinib PBPK model with optimized fm,CYP3A4 and fm,CYP2C9 was established to evaluate victim DDI risks. The previous minimal PBPK model was supported by the FDA for the dose reduction strategy, halving the dose with the concomitant use of strong CYP3A4 inhibitors and dual inhibitors on CYP2C9 and CYP3A4, such as fluconazole at ≤200 mg. Fluconazole simulation results were used as supportive evidence in discussions with the FDA and EMA about ruxolitinib dose adjustment when co-administering perpetrator drugs. Thus, this study demonstrated that PBPK modeling can support characterizing DDI liabilities to inform the drug label and might help reduce the number of clinical DDI studies by simulations of untested scenarios, when a robust PBPK model is established.


Coefficient of failure: a methodology for examining longitudinal beta-cell function in Type 2 diabetes.
We describe a new method, the determination of the coefficient of failure, which allows the assessment of beta-cell failure from any index of glycaemia. Previous methods using glycaemic thresholds and calculating time-to-failure have systematic deficiencies relating to bias, reproducibility and statistical power. Analyses using threshold methodologies and conventional survival analysis have an intrinsic disadvantage in that they use categorical data and thus make no allowance for near-failure, or progression towards failure. In contrast, the coefficient of failure includes all data in the analysis and takes into account improvement of glycaemia as well as deterioration of glycaemia. We describe the use of a 'coefficient of failure' defined as the slope of the least-squares regression line of a glycaemic index vs. time calculated for each individual patient on constant monotherapy. We exemplify the method using HbA1c levels from data from patients on chlorpropamide (n = 64) or glibenclamide (n = 65) monotherapy in the Oxford cohort of the UKPDS. Chlorpropamide-treated patients showed a mean coefficient of failure of 0.34 HbA(1c)%/year (0.44%/year sd) and glibenclamide-treated patients 0.50 HbA(1c)%/year (0.50%/year sd) (P = 0.046; unpaired two-tailed t-test). Kolmogorov-Smirnov testing demonstrated that the coefficients did not differ significantly from a normal distribution (chlorpropamide P = 0.12; glibenclamide P = 0.13). The coefficient of failure gives an estimate of beta-cell failure using any index of glycaemia. The coefficient is not constrained by predetermined glycaemic thresholds for failure and it allows the rate of decline in beta-cell function to be determined on any therapy or combination of therapies.


Incorporation of rhodamine B into male tobacco budworm moths Heliothis virescens to use as a marker for mating studies.
Rhodamine B, a dye commonly used in a variety of biological studies was incorporated into the bodies of male tobacco budworm moths, Heliothis virescens (Lepidoptera: Noctuidae), by allowing them to feed freely on 0.1% rhodamine dissolved in a 10% sucrose solution. After exposing males for one to three days to this pigment, rhodamine was clearly detectable in >82% of spermatophores extracted from untreated females. The intake of this dye did not affect the life span, the production of eggs or the capacity of moths to copulate when compared with moths fed only a sucrose solution or water. Rhodamine B was easily identifiable externally but was more apparent internally in males after only one day of exposure to the pigment. Even at this short feeding duration, rhodamine was detectable in >50% of males 5 days after feeding stopped. Longer exposure to the dye significantly increased the percentage stained. Detection of rhodamine was slightly enhanced by the use of ultraviolet light. The dye accumulation in internal abdominal organs was a better indicator of the presence of the pigment than external contamination of the moth. The use of the method described in this report can be a tool for the rapid incorporation of a low cost dye in the tobacco budworm for biological, behavioral and genetic studies.


Early phase Technology Assessment of nanotechnology in oncology.
To perform early Technology Assessment (TA) of nanotechnology in oncology. The possibilities of nanotechnology for detection (imaging), diagnosis and treatment of cancer are subject of different research programs where major investments are concerned. As a range of bio- nanotechnologies is expected to enter the oncology field it is relevant to consider the various aspects involved in especially early TA. This article provides two cases of early assessment of (predecessors of) nanotechnologies: Microarray Analysis and Photodynamic Therapy implementation, which methodology can be extrapolated to other nanotechnologies in oncology. Constructive Technology Assessment (CTA) is used for the introduction of technologies that are still in a dynamic phase of development or in an early stage of diffusion. The selection of studied aspects in CTA is based on: clinical aspects (safety, efficacy, and effectiveness), economic (cost-effectiveness), patient related (QoL, ethical/juridical and psychosocial), organizational aspects (diffusion and adoption) and scenario drafting. The features of the technology and the phase of implementation are decisive for choices and timing of the specific aspects to be studied. A framework was drafted to decide on the relevant aspects. In the first case, early implementation of Microarray Analysis; clinical effectiveness, logistics, patient centeredness and scenario drafting were given priority. Related to the diffusion-phase of Photodynamic Therapy however other aspects were evaluated, such as early cost-effectiveness analysis for possible reimbursement. Often CTA will result in a mixed method design. Especially scenario drafting is a powerful instrument to predict possible developments that can be anticipated upon in the assessment. CTA is appropriate for the study of early implementation of new technologies in oncology. In early TA small series often necessitate a mix of quantitative and qualitative methods. The features of nanotechnology involved are decisive for the selection of CTA aspects, most likely: safety -especially possible interactions with other technologies-, ethics, cost-effectiveness and patient centeredness.


The association between the upper digestive tract microbiota by HOMIM and oral health in a population-based study in Linxian, China.
Bacteria affect oral health, but few studies have systematically examined the role of bacterial communities in oral diseases. We examined this relationship in a large population-based Chinese cancer screening cohort. Human Oral Microbe Identification Microarrays were used to test for the presence of 272 human oral bacterial species (97 genera) in upper digestive tract (UDT) samples collected from 659 participants. Oral health was assessed using US NHANES (National Health and Nutrition Examination Survey) protocols. We assessed both dental health (total teeth missing; tooth decay; and the decayed, missing, and filled teeth (DMFT) score) and periodontal health (bleeding on probing (BoP) extent score, loss of attachment extent score, and a periodontitis summary estimate). Microbial richness, estimated by number of genera per sample, was positively correlated with BoP score (P = 0.015), but negatively correlated with tooth decay and DMFT score (P = 0.008 and 0.022 respectively). Regarding β-diversity, as estimated by the UniFrac distance matrix for pairwise differences among samples, at least one of the first three principal components of the UniFrac distance matrix was correlated with the number of missing teeth, tooth decay, DMFT, BoP, or periodontitis. Of the examined genera, Parvimonas was positively associated with BoP and periodontitis. Veillonellacease [G-1] was associated with a high DMFT score, and Filifactor and Peptostreptococcus were associated with a low DMFT score. Our results suggest distinct relationships between UDT microbiota and dental and periodontal health. Poor dental health was associated with a less microbial diversity, whereas poor periodontal health was associated with more diversity and the presence of potentially pathogenic species.


ER and PR signaling nodes during mammary gland development.
The ovarian hormones estrogen and progesterone orchestrate postnatal mammary gland development and are implicated in breast cancer. Most of our understanding of the molecular mechanisms of estrogen receptor (ER) and progesterone receptor (PR) signaling stems from in vitro studies with hormone receptor-positive cell lines. They have shown that ER and PR regulate gene transcription either by binding to DNA response elements directly or via other transcription factors and recruiting co-regulators. In addition they cross-talk with other signaling pathways through nongenomic mechanisms. Mouse genetics combined with tissue recombination techniques have provided insights about the action of these two hormones in vivo. It has emerged that hormones act on a subset of mammary epithelial cells and relegate biological functions to paracrine factors. With regards to hormonal signaling in breast carcinomas, global gene expression analyses have led to the identification of gene expression signatures that are characteristic of ERα-positive tumors that have stipulated functional studies of hitherto poorly understood transcription factors. Here, we highlight what has been learned about ER and PR signaling nodes in these different systems and attempt to lay out in which way the insights may converge.