New cardiac and skeletal protein aggregate myopathy associated with combined MuRF1 and MuRF3 mutations.
Protein aggregate myopathies (PAMs) define muscle disorders characterized by protein accumulation in muscle fibres. We describe a new PAM in a patient with proximal muscle weakness and hypertrophic cardiomyopathy, whose muscle fibres contained inclusions containing myosin and myosin-associated proteins, and aberrant distribution of microtubules. These lesions appear as intact A- and M-bands lacking thin filaments and Z-discs. These features differ from inclusions in myosin storage myopathy (MSM), but are highly similar to those in mice deficient for the muscle-specific RING finger proteins MuRF1 and MuRF3. Sanger sequencing excluded mutations in the MSM-associated gene MYH7 but identified mutations in TRIM63 and TRIM54, encoding MuRF1 and MuRF3, respectively. No mutations in other potentially disease-causing genes were identified by Sanger and whole exome sequencing. Analysis of seven family members revealed that both mutations segregated in the family but only the homozygous TRIM63 null mutation in combination with the heterozygous TRIM54 mutation found in the proband caused the disease phenotype. Both MuRFs are microtubule-associated proteins localizing to sarcomeric M-bands and Z-discs. They are E3 ubiquitin ligases that play a role in degradation of sarcomeric proteins, stabilization of microtubules and myogenesis. Lack of ubiquitin and the 20S proteasome subunit in the inclusions found in the patient suggested impaired turnover of thick filament proteins. Disruption of microtubules in cultured myotubes was rescued by transient expression of wild-type MuRF1. The unique features of this novel myopathy point to defects in homeostasis of A-band proteins in combination with instability of microtubules as cause of the disease.


Reduced chromatic discrimination in children with autism spectrum disorders.
Atypical perception in Autism Spectrum Disorders (ASD) is well documented (Dakin & Frith, 2005). However, relatively little is known about colour perception in ASD. Less accurate performance on certain colour tasks has led some to argue that chromatic discrimination is reduced in ASD relative to typical development (Franklin, Sowden, Burley, Notman & Alder, 2008). The current investigation assessed chromatic discrimination in children with high-functioning autism (HFA) and typically developing (TD) children matched on age and non-verbal cognitive ability, using the Farnsworth-Munsell 100 hue test (Experiment 1) and a threshold discrimination task (Experiment 2). In Experiment 1, more errors on the chromatic discrimination task were made by the HFA than the TD group. Comparison with test norms revealed that performance for the HFA group was at a similar level to typically developing children around 3 years younger. In Experiment 2, chromatic thresholds were elevated for the HFA group relative to the TD group. For both experiments, reduced chromatic discrimination in ASD was due to a general reduction in chromatic sensitivity rather than a specific difficulty with either red-green or blue-yellow subsystems of colour vision. The absence of group differences on control tasks ruled out an explanation in terms of general task ability rather than chromatic sensitivity. Theories to account for the reduction in chromatic discrimination in HFA are discussed, and findings are related to cortical models of perceptual processing in ASD.


Regions of the retinoblastoma gene product required for its interaction with the E2F transcription factor are necessary for E2 promoter repression and pRb-mediated growth suppression.
Studies of naturally occurring mutations of the RB1 tumor suppressor gene have indicated that the E1A/T antigen-binding domain is important for pRb function. Mutations engineered within the C-terminal 135 amino acids of pRb also abrogate its growth-suppressive function during the G1 interval of the cell cycle. Both the pRb E1A/T antigen-binding domain and the C-terminal domain are required for interaction with the E2F transcription factor. A series of mutated pRb proteins has been used to define the C-terminal sequences which determine E2F binding, adenovirus E2 promoter inhibition, and negative growth control. Deletion of the C terminus to residue 870 allowed full pRb function, while further deletion to residue 841 inactivated pRb in each assay. Amino acid sequences immediately C-terminal to the E1A/T antigen-binding domain were absolutely required for pRb activity. Mutations which prevented pRb from interacting with E2F also eliminated pRb-mediated E2 promoter repression and inactivated the ability of pRb to suppress cell growth.


Differing Methods and Definitions Influence DALY estimates: Using Population-Based Data to Calculate the Burden of Convulsive Epilepsy in Rural South Africa.
The disability adjusted life year (DALY) is a composite measure of disease burden that includes both morbidity and mortality, and is relevant to conditions such as epilepsy that can limit productive functioning. The 2010 Global Burden of Disease (GBD) study introduced a number of new methods and definitions, including a prevalence-based approach and revised disability weights to calculate morbidity and new standard life expectancies to calculate premature mortality. We used these approaches, and local, population-based data, to estimate the burden of convulsive epilepsy in rural South Africa. Comprehensive prevalence, incidence and mortality data on convulsive epilepsy were collected within the Agincourt sub-district in rural northeastern South Africa between 2008 and 2012. We estimated DALYs using both prevalence- and incidence-based approaches for calculating years of life lived with disability. Additionally, we explored how changing the disease model by varying the disability weights influenced DALY estimates. Using the prevalence-based approach, convulsive epilepsy in Agincourt resulted in 332 DALYs (95% uncertainty interval (UI): 216-455) and 4.1 DALYs per 1,000 individuals (95%UI: 2.7-5.7) annually. Of this, 26% was due to morbidity while 74% was due to premature mortality. DALYs increased by 10% when using the incidence-based method. Varying the disability weight from 0.072 (treated epilepsy, seizure free) to 0.657 (severe epilepsy) caused years lived with disability to increase from 18 (95%UI: 16-19) to 161 (95%UI: 143-170). DALY estimates are influenced by both the methods applied and population parameters used in the calculation. Irrespective of method, a significant burden of epilepsy is due to premature mortality in rural South Africa, with a lower burden than rural Kenya. Researchers and national policymakers should carefully interrogate the methods and data used to calculate DALYs as this will influence policy priorities and resource allocation.


Detection of misreported drug use in forensic populations: an overview of hair analysis.
Criminal and civil forensic evaluations are frequently complicated by misreported (denied or exaggerated) substance abuse. This article provides an introduction to the use of radioimmunoassay of hair (RIAH). RIAH with gas chromatography/mass spectrometry confirmation is superior to the more commonly employed urinalysis in its (1) wider window of detection, (2) ability to provide a chronology of substance abuse, and (3) resistance to countermeasures. The potential implications of the RIAH procedure to retrospective and prospective forensic evaluations are discussed.


MEIS2 Is an Oncogenic Partner in AML1-ETO-Positive AML.
Homeobox genes are known to be key factors in leukemogenesis. Although the TALE family homeodomain factor Meis1 has been linked to malignancy, a role for MEIS2 is less clear. Here, we demonstrate that MEIS2 is expressed at high levels in patients with AML1-ETO-positive acute myeloid leukemia and that growth of AML1-ETO-positive leukemia depends on MEIS2 expression. In mice, MEIS2 collaborates with AML1-ETO to induce acute myeloid leukemia. MEIS2 binds strongly to the Runt domain of AML1-ETO, indicating a direct interaction between these transcription factors. High expression of MEIS2 impairs repressive DNA binding of AML1-ETO, inducing increased expression of genes such as the druggable proto-oncogene YES1. Collectively, these data describe a pivotal role for MEIS2 in AML1-ETO-induced leukemia.


Endogenous glucose production is inhibited by the adipose-derived protein Acrp30.
Intraperitoneal injection of purified recombinant Acrp30 lowers glucose levels in mice. To gain insight into the mechanism(s) of this hypoglycemic effect, purified recombinant Acrp30 was infused in conscious mice during a pancreatic euglycemic clamp. In the presence of physiological hyperinsulinemia, this treatment increased circulating Acrp30 levels by approximately twofold and stimulated glucose metabolism. The effect of Acrp30 on in vivo insulin action was completely accounted for by a 65% reduction in the rate of glucose production. Similarly, glucose flux through glucose-6-phosphatase (G6Pase) decreased with Acrp30, whereas the activity of the direct pathway of glucose-6-phosphate biosynthesis, an index of hepatic glucose phosphorylation, increased significantly. Acrp30 did not affect the rates of glucose uptake, glycolysis, or glycogen synthesis. These results indicate that an acute increase in circulating Acrp30 levels lowers hepatic glucose production without affecting peripheral glucose uptake. Hepatic expression of the gluconeogenic enzymes phosphoenolpyruvate carboxykinase and G6Pase mRNAs was reduced by more than 50% following Acrp30 infusion compared with vehicle infusion. Thus, a moderate rise in circulating levels of the adipose-derived protein Acrp30 inhibits both the expression of hepatic gluconeogenic enzymes and the rate of endogenous glucose production.


Thematic analysis of personal statements in physician assistant program admissions.
The written personal statement is widely used in health professions program admissions. The purposes of this study were to identify the common themes manifesting in the personal statements of physician assistant (PA) program applicants and to measure the odds of matriculation while controlling for other admission covariates. This study was a retrospective mixed-method observational study of CASPA admissions data. From the aggregate pool of 14,682 CASPA applications in the 2009-2010 admissions cycle, we randomly selected a subset of 600 unique de-identified applicants with complete application data. We coded the major themes and subthemes for each personal statement. We then performed maximum likelihood logistic regression analysis that compared the odds of matriculation based on the major themes and known cognitive admission variables. We identified eight major themes including altruism and the desire to help people, challenges and hardships, experience, key accomplishments, personal characteristics, positive perception of PA career attributes, role models, and a religious or spiritual quest. The only major theme increasing the odds of matriculation was role models, specifically exposure to a PA role model. Grade-point average far exceeded all other variables influencing the odds of matriculation. In this study, we found the personal statement to be an unreliable tool for predicting successful PA program matriculation.


Assessing disproportionate costs to achieve good ecological status of water bodies in a Mediterranean river basin.
Water management is becoming increasingly important as the demand for water grows, diversifies, and includes more complex environmental concerns. The Water Framework Directive (WFD) seeks to achieve a good ecological status for all European Community water bodies by 2015. To achieve this objective, economic consideration of water management must be given to all decision-making processes. Exemption (time or level of stringency) from the objectives of the EU Directive can be justified by proving that the cost of implementing measures is disproportionate to the benefits. This paper addresses the issue of disproportionate costs through a cost-benefit analysis (CBA). To predict the costs, the function costs method is used. The quantification of environmental benefits is more complex, because they are not determined by the market. As an alternative to stated preference methods, we use the distance function approach to estimate the environmental benefits of improving water quality. We then apply this methodological approach to a Mediterranean River Basin in Spain. The results show that the achievement of good status could not be rejected based on the criterion of disproportionate costs in this river basin. This paper illustrates that CBA is a useful tool to inform policy and decision making. Furthermore, it is shown that economics, particularly the valuation of environmental benefits, plays a crucial role in fulfilling the environmental objectives of the WFD.


Bifunctional Ligands for Inhibition of Tight-Binding Protein-Protein Interactions.
The acknowledged potential of small-molecule therapeutics targeting disease-related protein-protein interactions (PPIs) has promoted active research in this field. The strategy of using small molecule inhibitors (SMIs) to fight strong (tight-binding) PPIs tends to fall short due to the flat and wide interfaces of PPIs. Here we propose a biligand approach for disruption of strong PPIs. The potential of this approach was realized for disruption of the tight-binding (KD = 100 pM) tetrameric holoenzyme of cAMP-dependent protein kinase (PKA). Supported by X-ray analysis of cocrystals, bifunctional inhibitors (ARC-inhibitors) were constructed that simultaneously associated with both the ATP-pocket and the PPI interface area of the catalytic subunit of PKA (PKAc). Bifunctional inhibitor ARC-1411, possessing a KD value of 3 pM toward PKAc, induced the dissociation of the PKA holoenzyme with a low-nanomolar IC50, whereas the ATP-competitive inhibitor H89 bound to the PKA holoenzyme without disruption of the protein tetramer.


Melatonin receptors, melatonin metabolizing enzymes and cyclin D1 in human breast cancer.
Melatonin suppresses breast cancer cell proliferation by inhibiting the upregulation of estrogen-induced cyclin D1 via its G-protein-coupled receptor MT1. Additionally, melatonin stimulates the expression of the estrogen sulfotransferase, SULT1E1. However, metabolism of melatonin via 6-hydroxylation by CYP1A1/1A2 and subsequent sulfonation by SULT1A1/1A3 decreases its intracellular concentration. This could have a negative impact on its oncostatic action in breast cancer. In this pilot study, we performed immunohistochemical (IHC) analysis of MT1 and cyclin D1 in breast cancer specimens from 33 patients. Also, we investigated the expression of CYP1A1/1A2, SULT1A1/1A3/1E1,and cyclin D1 in cancer (CANC) and adjacent non-cancer (NCANC) specimens from 10 representative breast cancer patients using quantitative real-time reverse transcription polymerase chain reaction. CYP1A1-mRNA-expression was found only in three CANC and in one NCANC. CYP1A2 mRNA was below the detection limit in all patients. SULT1A1 was observed only in two of the 10 CANC and one of the 10 NCANC specimens. But, all 10 CANC and NCANC samples showed high SULT1A3 levels. Cyclin D1 mRNA levels were found in all 10 CANC and NCANC specimens. Furthermore, IHC-staining of cyclin D1 was observed in 27 of 33 CANC and correlated positively with estrogen receptor positivity (p = 0.015). The low or even absent expression of CYP1A1 or CYP1A2 in breast cancer specimens suggested that melatonin might be involved in cell cycle arrest.


Examining intervention mechanisms of action using mediation analysis within a randomised trial of a whole-school health intervention.
Interventions to modify school environments are effective in promoting young people's health across outcomes, but mechanisms are poorly understood. We assessed mediation in a trial of the Learning Together intervention, building on the recent publication of results of effectiveness for reducing bullying and benefits across secondary outcomes and generally good implementation fidelity. Within a cluster-randomised trial involving 40 English schools, we examined student-reported and staff-reported school climate and student-reported involvement with delinquent peers at 24-month and 36-month follow-up, assessing the reliability of measures and whether these mediated health outcomes at a final follow-up. Response rates and reliability were good for student-reported but not staff-reported measures. The intervention increased student-reported but not staff-reported-positive school climate but, like effects on student health outcomes, these manifested only at a final follow-up. The intervention reduced student-reported contact with delinquent peers at an interim follow-up. Student-reported potential mediators measured at the interim follow-up were associated with most health outcomes at the final follow-up. Adjustment for student-reported school climate and contact with delinquent peers at the interim follow-up did not reduce the associations between trial arm and our health outcomes. Despite being constrained by imperfect measures and by the late manifestation of impacts on student-reported school climate undermining ability to assess mediation, our study for the first time provides tentative evidence that mediation of intervention effects via improved climate and disengagement from delinquent peers is plausible. Our study provides the first evidence from a trial that whole-school interventions may work by modifying school environments and student relationships. ISRCTN10751359.


Relationship between 17 Venezuelan counties estimated through communality of surnames.
The coefficients of relationship and the Euclidean distances between 17 Venezuelan counties were estimated based on the frequency distribution of surnames obtained from the 1984 Venezuelan register of electors. In general, the coefficients of relationship between counties within the same state were higher than those between counties from different states. Euclidean distances exhibited the opposite relationship. Spearman's correlation coefficients for 136 pairs of counties were estimated between geographic and Euclidean distances (r = 0.41), geographic distance and coefficient of relationship (r = -0.44) and between Euclidean distance and coefficient of relationship (r = -0.48). The effect of partial geographic isolation on the frequency distribution of surnames is shown in the State of Falcón, where an isthmus separates two counties of the peninsula from two others on the mainland, and in Mérida, where the Venezuelan Andes separates three counties from the rest of the country. Our results suggest that Euclidean distances are less influenced by common surnames than the coefficients of relationship. They also indicate that in Venezuela a high proportion of the population has remained sedentary until recently, and this gives rise to the focal distribution of some surnames.


Obesity in aging exacerbates blood-brain barrier disruption, neuroinflammation, and oxidative stress in the mouse hippocampus: effects on expression of genes involved in beta-amyloid generation and Alzheimer's disease.
There is growing evidence that obesity has deleterious effects on the brain and cognitive function in the elderly population. However, the specific mechanisms through which aging and obesity interact to promote cognitive decline remain unclear. To test the hypothesis that aging exacerbates obesity-induced cerebromicrovascular damage and neuroinflammation, we compared young (7 months) and aged (24 months) high fat diet-fed obese C57BL/6 mice. Aging exacerbated obesity-induced systemic inflammation and blood-brain barrier disruption, as indicated by the increased circulating levels of proinflammatory cytokines and increased presence of extravasated immunoglobulin G in the hippocampus, respectively. Obesity-induced blood-brain barrier damage was associated with microglia activation, upregulation of activating Fc-gamma receptors and proinflammatory cytokines, and increased oxidative stress. Treatment of cultured primary microglia with sera derived from aged obese mice resulted in significantly more pronounced microglia activation and oxidative stress, as compared with treatment with young sera. Serum-induced activation and oxidative stress were also exacerbated in primary microglia derived from aged animals. Hippocampal expression of genes involved in regulation of the cellular amyloid precursor protein-dependent signaling pathways, beta-amyloid generation, and the pathogenesis of tauopathy were largely unaffected by obesity in aged mice. Collectively, obesity in aging is associated with a heightened state of systemic inflammation, which exacerbates blood-brain barrier disruption. The resulting neuroinflammation and oxidative stress in the mouse hippocampus likely contribute to the significant cognitive decline observed in aged obese animals.


IL-13-induced airway mucus production is attenuated by MAPK13 inhibition.
Increased mucus production is a common cause of morbidity and mortality in inflammatory airway diseases, including asthma, chronic obstructive pulmonary disease (COPD), and cystic fibrosis. However, the precise molecular mechanisms for pathogenic mucus production are largely undetermined. Accordingly, there are no specific and effective anti-mucus therapeutics. Here, we define a signaling pathway from chloride channel calcium-activated 1 (CLCA1) to MAPK13 that is responsible for IL-13-driven mucus production in human airway epithelial cells. The same pathway was also highly activated in the lungs of humans with excess mucus production due to COPD. We further validated the pathway by using structure-based drug design to develop a series of novel MAPK13 inhibitors with nanomolar potency that effectively reduced mucus production in human airway epithelial cells. These results uncover and validate a new pathway for regulating mucus production as well as a corresponding therapeutic approach to mucus overproduction in inflammatory airway diseases.


[Infectious keratitis in contact lens wearers].
Infectious keratitis in contact lens wearers Infectious keratitis is a sight-threatening complication in contact lens wearers. The infection is most frequently caused by bacteria (Pseudomonas aeruginosa). However, fungi or Acanthamoeba are found in increasing frequency. Three cases illustrate a characteristic course: patient A (11-year-old male) was treated for three weeks before the characteristic aspect of Acanthamoeba keratitis was recognized and confirmed. Patient B (45-year-old female) developed a severe corneal ulcer within 4 days; microbiological diagnostics confirmed Pseudomonas aeruginosa keratitis. Examination of patient C (27-year-old female) showed an infiltrate with satellites, typical of fungal keratitis. It is important to check the use of contact lenses in patients with keratitis. Referral to the ophthalmologist is mandatory: immediate in cases with an infiltrate. A dentritiform epithelial lesion in a contact lens wearer is indicative of Acanthamoeba keratitis, whereas fungal keratitis shows satellites or feathering edges. Steroids may only be prescribed by an ophthalmologist after confirmation of the causative agent.


Effect of exclusion diet with nutraceutical therapy in juvenile Crohn's disease.
Most moderate-severe juvenile Crohn's disease (CD) patients are in a constant catabolic state resulting in poor weight gain and growth failure. Anti-inflammatory, immunomodulatory, and monoclonal antibody drugs, as well as growth hormone (GH), frequently fail to achieve sustained remission or reverse growth failure. To test whether an exclusion diet with nutraceutical therapy (DNT) could induce sustained clinical remission and weight gain, and if so does this enhance the ability for GH to reverse growth failure. An uncontrolled prospective case study was undertaken in six moderate- severe CD patients, two of whom had completed growth. All were treated with DNT. Adequate caloric and protein ( >or= 3g/kg/d) intake for catch up weight was prescribed. Dairy products, certain grains and carrageenan containing foods were eliminated. Nutraceuticals, consisting of fish peptides, bovine colostrum, boswellia serrata, curcumin and a multivitamin were administered daily. Lactobacillus GG, a probiotic, was administered twice weekly. Recombinant human GH (rhGH) was administered daily. Within 2 months of starting DNT all six patients went into remission, with discontinuation of all pharmacological drugs. Three patients have remained in sustained remission for 4 to 8 years. One patient with very severe CD had recurrence of CD symptoms after being in complete remission for 18 months, one patient was in remission for 3 years but symptoms recurred when she became less compliant to DNT and one recently treated patient remains in remission after 6 months. With the addition of rhGH, the 4 growing patients had good-excellent growth response DNT engendered prolonged remission and restoration of normal weight in moderate-severe juvenile CD patients, providing conditions that enabled rhGH to stimulate growth. These findings justify larger controlled trials to evaluate the long-term benefit of compliance to DNT in both juvenile and adult CD patients.


Optimal behavior: can foragers balance two conflicting demands?
According to much current theory, organisms should be able to balance the conflicting demands of the need to feed efficiently and the need to avoid preadtors while feeding. In an experimental conflict situation, it was possible to evaluate the relative fitnesses associated with the available choices and to compare the observed behaviors with predictions derived from fitness considerations. A backswimmer, Notonecta hoffmanni, was capable of balancing these two conflicting factors adaptively.


Development of guidelines to facilitate improved support of South Asian carers by primary health care teams.
Evidence based guidelines are regarded as an appropriate basis for providing effective health care, but few guidelines incorporate the views of users such as carers. To develop guidelines to assist primary health care teams (PHCTs) in their work with carers within South Asian communities. The guidelines were drawn up by a development group consisting of members of teams in areas with South Asian communities (Leicester and Bradford). The teams were invited to make their recommendations based on a systematic review of literature on minority ethnic carers and the findings of a study of the needs and experiences of local South Asian carers. A grading system was devised to enable the teams and a group of expert peer reviewers to assess the quality of evidence in support of each recommendation. The teams agreed seven recommendations, graded according to available evidence and strength of opinion. External peer review supported the PHCTs' interpretation of evidence and their recommendations. The recommendations included consideration of communication and information for carers, coordination of care within teams, and recognition by team members of the roles of carers and their cultural and religious beliefs. There are particular steps that PHCTs can take to improve their support of South Asian carers. It is possible to develop guidelines that take users' views into account and incorporate evidence from qualitative studies.


A decrease in vitamin D levels is associated with methotrexate-induced oral mucositis in children with acute lymphoblastic leukemia.
Children with acute lymphoblastic leukemia (ALL) are at increased risk of vitamin D deficiency, which might make them more susceptible to developing adverse events. Previous studies showed that low vitamin D levels were associated with an increased inflammatory mucosal state and impaired mucosal tissue barriers. We examined the prevalence of vitamin D deficiency and studied the association between vitamin D levels and methotrexate (MTX)-induced oral mucositis in pediatric ALL. We assessed 25-hydroxyvitamin D (25(OH)D3) and 24,25-dihydroxyvitamin D (24,25(OH)2D3) levels in 99 children with ALL before the start of 4 × 5 g/m2 high-dose methotrexate (HD-MTX) (T0) and in 81/99 children after discontinuation of HD-MTX (T1). Two cutoff values for vitamin D deficiency exist: 25(OH)D3 levels < 30 and < 50 nmol/L. Oral mucositis was defined as grade ≥ 3 according to the National Cancer Institute Criteria. Vitamin D deficiency occurred in respectively 8% (< 30 nmol/L) and 33% (< 50 nmol/L) of the patients at T0, and more frequently in children > 4 years of age as compared to children between 1 and 4 years of age. A decrease in 25(OH)D3 levels during HD-MTX therapy was associated with developing severe oral mucositis (OR 1.6; 95% CI [1.1-2.4]). 25(OH)D3 and 24,25(OH)2D3 levels at T0 and the change in 24,25(OH)2D3 levels during therapy were not associated with the development of severe oral mucositis. This study showed that vitamin D deficiency occurs frequently in pediatric ALL patients above the age of 4 years. A decrease in 25(OH)D3 levels during MTX therapy was observed in children with ALL that developed severe oral mucositis.


A novel construction of genome space with biological geometry.
A genome space is a moduli space of genomes. In this space, each point corresponds to a genome. The natural distance between two points in the genome space reflects the biological distance between these two genomes. Currently, there is no method to represent genomes by a point in a space without losing biological information. Here, we propose a new graphical representation for DNA sequences. The breakthrough of the subject is that we can construct the moment vectors from DNA sequences using this new graphical method and prove that the correspondence between moment vectors and DNA sequences is one-to-one. Using these moment vectors, we have constructed a novel genome space as a subspace in R(N). It allows us to show that the SARS-CoV is most closely related to a coronavirus from the palm civet not from a bird as initially suspected, and the newly discovered human coronavirus HCoV-HKU1 is more closely related to SARS than to any other known member of group 2 coronavirus. Furthermore, we reconstructed the phylogenetic tree for 34 lentiviruses (including human immunodeficiency virus) based on their whole genome sequences. Our genome space will provide a new powerful tool for analyzing the classification of genomes and their phylogenetic relationships.


Metalloproteinase-dependent and -independent processes contribute to inhibition of breast cancer cell migration, angiogenesis and liver metastasis by a disintegrin and metalloproteinase with thrombospondin motifs-15.
The ADAMTS proteinases are a family of secreted, matrix-associated enzymes that have diverse roles in the regulation of tissue organization and vascular homeostasis. Several of the 19 human family members have been identified as having either tumor promoting or suppressing roles. We previously demonstrated that decreased ADAMTS15 expression correlated with a worse clinical outcome in mammary carcinoma (e.g., Porter et al., Int J Cancer 2006;118:1241-7). We have explored the effects of A Disintegrin and Metalloproteinase with Thrombospondin motifs-15 (ADAMTS-15) on the behavior of MDA-MB-231 and MCF-7 breast cancer cells by stable expression of either a wild-type (wt) or metalloproteinase-inactive (E362A) protein. No effects on mammary cancer cell proliferation or apoptosis were observed for either form of ADAMTS-15. However, both forms reduced cell migration on fibronectin or laminin matrices, though motility on a Type I collagen matrix was unimpaired. Knockdown of syndecan-4 attenuated the inhibitory effects of ADAMTS-15 on cell migration. In contrast to its effects on cell migration, wt ADAMTS-15 but not the E362A inactive mutant inhibited endothelial tubulogenesis in 3D collagen gels and angiogenesis in the aortic ring assay. In experimental metastasis assays in nude mice, MDA-MB-231 cells expressing either form of ADAMTS-15 showed reduced spread to the liver, though lung colonization was enhanced for cells expressing wt ADAMTS-15. These studies indicate that extracellular ADAMTS-15 has multiple actions on tumor pathophysiology. Via modulation of cell-ECM interactions, which likely involve syndecan-4, it attenuates mammary cancer cell migration independent of its metalloproteinase activity; however, its antiangiogenic action requires catalytic functionality, and its effects on metastasis in vivo are tissue niche-dependent.


Feature Selection Algorithms Enhance the Accuracy of Frailty Indexes as Measures of Biological Age.
Biological age captures some of the variance in life expectancy for which chronological age is not accountable, and it quantifies the heterogeneity in the presentation of the aging phenotype in various individuals. Among the many quantitative measures of biological age, the mathematically uncomplicated frailty/deficit index is simply the proportion of the total health deficits in various health items surveyed in different individuals. We used 3 different statistical methods that are popular in machine learning to select 17-28 health items that together are highly predictive of survival/mortality, from independent study cohorts. From the selected sets, we calculated frailty indexes and Klemera-Doubal's biological age estimates, and then compared their mortality prediction performance using Cox proportional hazards regression models. Our results indicate that the frailty index outperforms age and Klemera-Doubal's biological age estimates, especially among the oldest old who are most prone to biological aging-caused mortality. We also showed that a DNA methylation index, which was generated by applying the frailty/deficit index calculation method to 38 CpG sites that were selected using the same machine learning algorithms, can predict mortality even better than the best performing frailty index constructed from health, function, and blood chemistry.


Reconstructed ancestral sequences improve pathogen identification using resequencing DNA microarrays.
We describe the benefit of using reconstructed ancestral sequences (RAS) on resequencing microarrays for rapid pathogen identification, with Enterobacteriaceae rpoB sequences as a model. Our results demonstrate a sharp improvement of call rate and accuracy when using RASs as compared to extant sequences. This improvement was attributed to the lower sequence divergence of RASs, which also expanded the sequence space covered by the microarray. Extension of this novel microarray design strategy to viruses, antimicrobial resistance elements or toxins is straightforward.


Enhanced amyloidogenic metabolism of the amyloid beta-protein precursor in the X11L-deficient mouse brain.
X11L, a neuronal adaptor protein, associates with the cytoplasmic domain of APP and suppresses APP cellular metabolism. APP is the precursor of Abeta, whose metabolism is strongly implicated in Alzheimer disease pathogenesis. To examine the roles of X11L function in APP metabolism, including the generation of Abeta in the brain, we produced X11L-deficient mutant mice on the C57BL/6 background. The mutant mice did not exhibit histopathological alterations or compensatory changes in the expression of other X11 family proteins, X11 and X11L2. The expression level and distribution of APP in the brain of mutant mice were also identical to those in wild-type mice. However, in the hippocampus, where substantial levels of X11L and APP are expressed, the mutant mice exhibited a significant increase in the level of the C-terminal fragments of APP produced by cleavage with beta-secretase but not alpha-secretase. The levels of Abeta were increased in the hippocampus of aged mutant mice as compared with age-matched controls. These observations clearly indicate that X11L suppresses the amyloidogenic but not amyloidolytic processing of APP in regions of the brain such as the hippocampus, which express significant levels of X11L.


Acute and persistent effects of pre- and posthatching thermal environments on growth and metabolism in the red-eared slider turtle, Trachemys scripta elegans.
Many ectotherms possess the capacity to survive a wide range of thermal conditions. Long-term exposure to temperature can induce acclimational and/or organizational effects, and the developmental stage at which temperature exposure occurs may affect the type, degree, and persistence of these effects. We incubated red-eared slider turtle embryos at three different constant temperatures (T(inc); 26.5, 28.5, 30.5°C), then divided the resulting hatchlings between two water temperatures (T(water); 25, 30°C). We calculated growth rates to assess the short- and long-term effects of thermal experience on this metabolically costly process. We also measured resting metabolic rate (RMR) at three body temperatures (T(body;) 26.5, 28.5, 30.5°C) shortly after hatching and 6 months posthatching to characterize the degree and persistence of acclimation to T(inc) and T(water) . Hatchling RMRs were affected by T(body) and T(inc) , and fit a pattern consistent with positive but incomplete metabolic compensation to T(inc) . Average growth rates over the first 11 weeks posthatching were strongly affected by T(water) but only marginally influenced by T(inc) , and only at T(water) = 30°C. Six-month RMRs exhibited strong acclimation to T(water) consistent with positive metabolic compensation. However, within each T(water) treatment, RMR fit patterns indicative of inverse metabolic compensation to T(inc) , opposite of the pattern observed in hatchlings. Average growth rates calculated over 6 months continued to show a strong effect of T(water) , and the previously weak effect of T(inc) observed within the 30°C T(water) treatment became more pronounced. Our results suggest that metabolic compensation was reversible regardless of the life stage during which exposure occurred, and therefore is more appropriately considered acclimational than organizational.


Staying Current: Developing Just-in-time Evidence-Based Learning Objectives for a Maternal Cardiac Arrest Simulation Curriculum.
Our objective was to review the latest evidence on resuscitation care for maternal cardiac arrest (MCA) and gain expert consensus on best practices to inform an evidence-based curriculum. We convened a multidisciplinary panel of stakeholders in MCA to develop an evidence-based simulation training, Obstetric Life Support™ (OBLS). To inform the learning objectives, we used a novel three-step process to achieve consensus on best practices for maternal resuscitation. First, we reaffirmed the evidence process on an existing MCA guideline using the Appraisal of Guidelines for Research and Evaluation (AGREE II). Next, via systematic review, we evaluated the latest evidence on MCA and identified emerging topics since the publication of the MCA guideline. Finally, we applied a modified Research and Development (RAND) technique to gain consensus on emerging topics to include as additional just-in-time best practices. The AGREE II survey results demonstrated unanimous consensus on reaffirmation of the 2015 American Heart Association (AHA) MCA guideline for inclusion into the OBLS curriculum. A systematic review with deduplication resulted in 11,871 articles for review. After categorizing and synthesizing the relevant literature, we presented twelve additional best practices to the expert panel using a modified RAND technique. Upon completion, the 2015 AHA statement and nine additional just-in-time best practices were affirmed to inform the OBLS curriculum. A novel three-step process including reaffirmation of evidence process, systematic review, and a modified RAND technique resulted in unanimous consensus from experts in MCA resuscitation on existing and new just-in-time best practices to inform the learning objectives for an evidence-based curriculum.


Fine definition of the pedigree haplotypes of closely related rice cultivars by means of genome-wide discovery of single-nucleotide polymorphisms.
To create useful gene combinations in crop breeding, it is necessary to clarify the dynamics of the genome composition created by breeding practices. A large quantity of single-nucleotide polymorphism (SNP) data is required to permit discrimination of chromosome segments among modern cultivars, which are genetically related. Here, we used a high-throughput sequencer to conduct whole-genome sequencing of an elite Japanese rice cultivar, Koshihikari, which is closely related to Nipponbare, whose genome sequencing has been completed. Then we designed a high-throughput typing array based on the SNP information by comparison of the two sequences. Finally, we applied this array to analyze historical representative rice cultivars to understand the dynamics of their genome composition. The total 5.89-Gb sequence for Koshihikari, equivalent to 15.7 x the entire rice genome, was mapped using the Pseudomolecules 4.0 database for Nipponbare. The resultant Koshihikari genome sequence corresponded to 80.1% of the Nipponbare sequence and led to the identification of 67,051 SNPs. A high-throughput typing array consisting of 1917 SNP sites distributed throughout the genome was designed to genotype 151 representative Japanese cultivars that have been grown during the past 150 years. We could identify the ancestral origin of the pedigree haplotypes in 60.9% of the Koshihikari genome and 18 consensus haplotype blocks which are inherited from traditional landraces to current improved varieties. Moreover, it was predicted that modern breeding practices have generally decreased genetic diversity Detection of genome-wide SNPs by both high-throughput sequencer and typing array made it possible to evaluate genomic composition of genetically related rice varieties. With the aid of their pedigree information, we clarified the dynamics of chromosome recombination during the historical rice breeding process. We also found several genomic regions decreasing genetic diversity which might be caused by a recent human selection in rice breeding. The definition of pedigree haplotypes by means of genome-wide SNPs will facilitate next-generation breeding of rice and other crops.


Profiling Detection and Classification of Lameness Methods in British Dairy Cattle Research: A Systematic Review and Meta-Analysis.
Lameness is a serious concern in the dairy sector, reflecting its high incidence and impact on animal welfare and productivity. Research has provided figures on its frequency using different methodologies, making it difficult to compare results and hindering farm-level decision-making. The study's objectives were to determine the frequency levels of lameness in British dairy cattle through a meta-analysis approach, and to understand the chronological patterns of how lameness cases are detected and classified in scientific research. A systematic review was conducted using PRISMA-P guidelines for article selection. Random-effects models estimated the pooled frequency measure of lameness with heterogeneity managed through subgroup analysis and meta-regression. Sixty-eight papers were identified, 50 included prevalence and 36 incidence data. The pooled prevalence of lameness in British dairy cattle was estimated at 29.5% (95% CI 26.7-32.4%) whilst all-cause lameness incidence rate indicated 30.9 cases of lameness per 100 cow-years (95% CI 24.5-37.9). The pooled cause-specific lameness incidence rate per 100 cow-years was 66.1 (95% CI 24.1-128.8) for white line disease, 53.2 (95% CI 20.5-101.2) for sole ulcer, 53.6 (95% CI 19.2-105.34) for digital dermatitis, with 51.9 (95% CI 9.3-129.2) attributable to other lameness-related lesions. Heterogeneity levels remained high. Sixty-nine papers contributed to a chronological overview of lameness data source. Although the AHDB Dairy mobility scoring system (MSS) was launched in the UK in 2008 and adopted shortly after by the British Dairy sector as the standard tool for assessing lameness, other methods are used depending on the investigator. Automated lameness detection systems may offer a solution for the subjective nature of MSSs, yet it was utilized in one study only. Despite the recognition of under-reporting of lameness from farm records 22 (31.9%) studies used this data source. The diversity of lameness data collection methods and sources was a key finding. It limits the understanding of lameness burden and the refinement of policy making for lameness. Standardizing case definition and research methods would improve knowledge of and ability to manage lameness. Regardless of the measurement method lameness in British dairy cattle is high.


Effects of epidermal growth factor-loaded mucoadhesive films on wounded oral tissue rafts.
Current treatments for traumatic oral mucosal wounds include the gold standard of autologous tissue and alternative tissue-engineered grafts. While use of autografts has disadvantages of minimal availability of oral keratinized tissue, second surgery, and donor site discomfort, tissue-engineered grafts are limited by their unavailability as off-the-shelf products owing to their fabrication time of 4-8 weeks. Hence, the current work aimed to develop a potentially cost-effective, readily available device capable of enhancing native mucosal regeneration. Considering the key role of epidermal growth factor (EGF) in promoting mucosal wound regeneration and the advantages of mucoadhesive delivery systems, mucoadhesive films composed of polyvinylpyrrolidone and carboxymethylcellulose were developed to provide sustained release of EGF for a minimum of 6 h. Bioactivity of released EGF supernatants was then confirmed by its ability to promote proliferation of BALB/3T3 fibroblasts. Efficacy of the developed system was then investigated in vitro using buccal tissues (ORL 300-FT) as a potential replacement for small animal studies. Although the mucoadhesive films achieved their desired role of delivering bioactive EGF in a sustained manner, treatment with EGF, irrespective of its release from the films or solubilized in medium, caused a hyperparakeratotic response from in vitro tissues with distinguishable histological features including thickening of the spinous layer, intra- and intercellular edema, and pyknotic nuclei. These significant morphological changes were associated with no improvements in wound closure. These observations raise questions about the potential of using in vitro tissues as a wound healing model and substitute for small animal studies. The mucoadhesive delivery system developed, however, with its potential for sustained release of bioactive growth factors and small molecules, may be loaded with other desired compounds, with or without EGF, to accelerate the process of wound healing.


[Causal inference in medicine--decision making].
In the field of occupational medicine, either when we consider some preventive plans or when we make decisions to compensate for occupational diseases, it has been necessary to discuss causality between work and disease. Furthermore, epidemiologic causality has recently been used in risk assessment in occupational and environmental settings. We have shown that the law of causality in medicine is recognized as probability and continuous variables. Such a law of causality has been recognized in the same way as probability in physics, too, and has been regarded as a model of science. Physicists and mathematicians had claimed the importance of probability in causal inference as well as the principle of uncertainty before it was discovered. We, then, explained Etiologic Fraction (EF), Attributable Proportion for the Exposed Population (APE), Probability of Causation (PC), and so on. The PC has been used to ascertain the conditional probability in an individual case of a disease having been caused by a particular prior exposure, by using the experience of exposed populations to determine the appropriate relative risk, and this has been used for compensation for exposed cases. Next the applicability of information from a population to individuals was presented. Third, we provided a brief historical aspect of epidemiology. The evolutions in Epidemiology have been very rapid, so we pointed out that, in Japan, we could observe many incommensurable phenomena in epidemiologists and physicians depending on the era which was studied by them. Fourth, we discussed judgement and political application based on epidemiologic evidence, using Yanagimoto's classification is also taken or not should be estimated and compared. We presented several examples of reasoning in judgements. Lastly, we discussed several tasks and assignments for the future of epidemiology.


Unexpected but Incidental Positive Outcomes Predict Real-World Gambling.
Positive mood can affect a person's tendency to gamble, possibly because positive mood fosters unrealistic optimism. At the same time, unexpected positive outcomes, often called prediction errors, influence mood. However, a linkage between positive prediction errors-the difference between expected and obtained outcomes-and consequent risk taking has yet to be demonstrated. Using a large data set of New York City lottery gambling and a model inspired by computational accounts of reward learning, we found that people gamble more when incidental outcomes in the environment (e.g., local sporting events and sunshine) are better than expected. When local sports teams performed better than expected, or a sunny day followed a streak of cloudy days, residents gambled more. The observed relationship between prediction errors and gambling was ubiquitous across the city's socioeconomically diverse neighborhoods and was specific to sports and weather events occurring locally in New York City. Our results suggest that unexpected but incidental positive outcomes influence risk taking.


Pre-operative distraction lengthening for radial longitudinal deficiency.
Patients treated for total radial aplasia have been reviewed. It was possible to re-align the carpus by radialization in five out of six limbs treated with pre-operative distraction, but in only one out of six treated without distraction. The average improvement in radial angulation in the non-distraction group was 19 degrees and in the distraction group 38 degrees. The average improvement in radial translation was 16 mm and 17 mm in the two groups respectively. Pre-operative distraction with the Kessler device permits re-alignment of the hand without skeletal resection or excessive tension on the radial neurovascular structures.


Bilateral percutaneous nephrolithotomy in a patient with hemophilia A disorder.
A 50-year-old man with hemophilia A presented with recurrent hematuria due to renal stone disease. He was receiving approximately 50,000 units of recombinant factor (rF) VIII concentrate every year due to hematuria. Between 1996 and 2002, his serum creatinine level increased from 0.7 to 1.2 ng/ml. In an effort to resolve the problems of excessive blood loss with transfusions, recurrent rF VIII replacements and deteriorating renal function, he was offered treatment with percutaneous nephrolithotomy (PNL) in conjunction with rF VIII administration. He underwent left PNL for left staghorn calculi in November 2002 with administration of 52,000 units rF VIII, and another PNL for the right kidney in April 2004 with the administration of 90,500 units rF VIII. A pneumatic lithotriptor was used in both operations. The serum creatinine level was 0.8 ng/ml upon completion of treatment and the patient was symptom and stone free at 10-month follow-up. He has not suffered from hematuria since that time. We conclude that bleeding disorders may not be a contraindication for PNL if corrected and monitored appropriately.


Physical Activity among Veterans and Nonveterans with Diabetes.
Engaging in regular physical activity (PA), with or without a corresponding decrease in weight, is associated with improved health outcomes. The purpose of this study was to quantify the extent to which PA differed between veterans and nonveterans and to determine how diabetes and age influenced this association. Data from the 2009 Behavioral Risk Factor Surveillance System were used in this study. Respondents were classified as having diabetes if they reported ever being diagnosed with diabetes except during pregnancy. Respondents who reported ever serving on active military duty were classified as veterans. Based on self-report, we calculated the average minutes per week of moderate, vigorous, and total activity. After adjusting for sex, race and ethnicity, household income, education level, body mass index (BMI), and recent health checkup, veteran status was associated with a small but significantly larger amount of average weekly moderate PA (2.2 minutes, P = 0.0058) but not average weekly vigorous PA (-0.02 minutes, P = 0.98). Diabetes and prediabetes were associated with significantly lower mean levels of both moderate and vigorous intensity PA, as was increasing age. Consistent with prior research, veterans engaged in more PA than nonveterans. The association between diabetes, age, and physical activity did not differ by veteran status.


Quality of life assessed with EQ-5D in patients undergoing glioma surgery: what is the responsiveness and minimal clinically important difference?
To evaluate the responsiveness of EQ-5D 3L in patients undergoing intracranial glioma surgery and estimate the minimal clinically important difference (MCID). EQ-5D 3L index values from 164 patients who underwent glioma surgery in the period 2007-2012 were analysed. Responsiveness and MCID were estimated using a combination of distribution-based and anchor-based methods. Karnofsky performance status served as an anchor. Patients who improved functionally did not report significantly higher EQ-5D 3L scores post operatively with a standardized response mean (SRM) of 0.04 (p = 0.13). Patients who deteriorated functionally reported significantly lower EQ-5D 3L scores post operatively with a SRM of 0.72 (p < 0.001). With different approaches, we determined a range of MCID values from 0.13 to 0.15. EQ-5D 3L is responsive to changes when glioma patients are deteriorating functionally after surgery but not responsive when the patients are improving. The MCID values for EQ-5D 3L in glioma surgery seem higher than reported MCID values for other types of cancers.


Emergence of a novel GII.17 norovirus – End of the GII.4 era?
In the winter of 2014/15 a novel GII.P17-GII.17 norovirus strain (GII.17 Kawasaki 2014) emerged, as a major cause of gastroenteritis outbreaks in China and Japan. Since their emergence these novel GII.P17-GII.17 viruses have replaced the previously dominant GII.4 genotype Sydney 2012 variant in some areas in Asia but were only detected in a limited number of cases on other continents. This perspective provides an overview of the available information on GII.17 viruses in order to gain insight in the viral and host characteristics of this norovirus genotype. We further discuss the emergence of this novel GII.P17-GII.17 norovirus in context of current knowledge on the epidemiology of noroviruses. It remains to be seen if the currently dominant norovirus strain GII.4 Sydney 2012 will be replaced in other parts of the world. Nevertheless, the public health community and surveillance systems need to be prepared in case of a potential increase of norovirus activity in the next seasons caused by this novel GII.P17-GII.17 norovirus.


Biogenic transport of glyphosate in the presence of LDPE microplastics: A mesocosm experiment.
The accumulation of plastic debris and herbicide residues has become a huge challenge and poses many potential risks to environmental health and soil quality. In the present study, we investigated the transport of glyphosate and its main metabolite, aminomethylphosphonic acid (AMPA) via earthworms in the presence of different concentrations of light density polyethylene microplastics in the litter layer during a 14-day mesocosm experiment. The results showed earthworm gallery weight was negatively affected by the combination of glyphosate and microplastics. Glyphosate and AMPA concentrated in the first centimetre of the top soil layer and the downward transport of glyphosate and AMPA was only detected in the earthworm burrows, ranging from 0.04 to 4.25 μg g-1 for glyphosate and from 0.01 (less than limit of detection) to 0.76 μg g-1 for AMPA. The transport rate of glyphosate (including AMPA) from the litter layer into earthworm burrows ranged from 6.6 ± 4.6% to 18.3 ± 2.4%, depending on synergetic effects of microplastics and glyphosate application. The findings imply that earthworm activities strongly influence pollutant movement into the soil, which potentially affects soil ecosystems. Further studies focused on the fate of pollutants in the microenvironment of earthworm burrows are needed.


Hypolipidaemia among patients with PMM2-CDG is associated with low circulating PCSK9 levels: a case report followed by observational and experimental studies.
Proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors are novel therapeutics for reducing low-density lipoprotein cholesterol (LDLc). While serious side-effects have not been observed in short-term clinical trials, there remain concerns that long-term PCSK9 inhibition may cause neurocognitive side-effects. An adult male with childhood-onset global developmental delay, cerebellar atrophy and severe hypolipidaemia underwent extensive biochemical and genetic investigations. Initial testing revealed low circulating PCSK9 levels and a common loss-of-function PCSK9 polymorphism, but these findings did not fully account for severe hypolipidaemia. Whole-exome sequencing was subsequently performed and identified two pathogenic phosphomannose mutase 2 (PMM2) variants (p.Arg141His and p.Pro69Ser) known to cause PMM2-associated congenital disorder of glycosylation (PMM2-CDG). A diagnosis of PMM2-CDG was consistent with the proband's neurological symptoms and severe hypolipidaemia. Given that PMM2-CDG is characterised by defective protein N-glycosylation and that PCSK9 is a negative regulator of LDLc, we postulated that loss of PCSK9 N-glycosylation mediates hypolipidaemia among patients with PMM2-CDG. First, in an independent cohort of patients with PMM2-CDG (N=8), we verified that circulating PCSK9 levels were significantly lower in patients than controls (p=0.0006). Second, we conducted in vitro experiments in hepatocyte-derived cells to evaluate the effects of PCSK9 N-glycosylation loss on LDL receptor (LDLR) activity. Experimental results suggest that defective PCSK9 N-glycosylation reduces the ability of circulating PCSK9 to degrade LDLR. Life-long exposure to genetically lower PCSK9 per se is unlikely to cause neurocognitive impairment. Both observational and experimental findings suggest that hypolipidaemia in PMM2-CDG may be partially mediated by loss of PCSK9 N-glycosylation and/or its regulators.


Decompressive craniectomy in children: single-center series and systematic review.
Decompressive craniectomy (DC) is performed as a life-saving procedure in patients with intractably increased intracranial pressure after traumatic brain injury, bleeding, cerebral infarction, or brain swelling of other causes. However, the application of DC is as controversial in the pediatric population as it is in adults. To find factors influencing the outcome in pediatric patients who underwent DC because of sustained high intracranial pressure. Between April 2000 and December 2009, 34 pediatric patients (age 0-18 years) underwent DC. Patients were stratified according to the indication for DC. Outcome was assessed according to the modified Rankin Scale score at 6 months. MEDLINE was searched for published studies or reports of DC in pediatric patients to gain a larger population. Two reviewers independently extracted data. Literature data, including the current series, revealed a total of 172 pediatric patients. Overall, a favorable outcome was achieved in 106 of 172 patients (62%). A favorable outcome was achieved in 25 of 36 patients without traumatic brain injury vs 81 of 136 patients with traumatic brain injury (69% vs 60%). Patients without signs of cerebral herniation had a better outcome than patients with unilateral or bilateral dilated pupils (73% vs 60% vs 45%, respectively). The current data indicate that DC in children with traumatic or nontraumatic brain swelling might be warranted, regardless of the underlying cause. Despite mydriasis, a favorable outcome might be achieved in a significant number of pediatric patients. Nevertheless, careful individual decision making is needed for each patient, especially when signs of cerebral herniation have persisted for a long time.


Once-weekly rifapentine and isoniazid for tuberculosis prevention in patients with HIV taking dolutegravir-based antiretroviral therapy: a phase 1/2 trial.
Short-course preventive therapy with 12 doses of once-weekly rifapentine (900 mg) plus isoniazid (900 mg) could greatly improve tuberculosis control, especially in areas with high co-endemicity with HIV. However, a small previous trial of such therapy with dolutegravir in healthy, HIV-negative adults was halted early after two of the four patients developed serious adverse events. Because of the potential use of this therapy, and variable safety outcomes of tuberculosis drugs seen in patients with and without HIV, we aimed to characterise safety, pharmacokinetics, and virological suppression in adults who are HIV positive. DOLPHIN was a phase 1/2, single-arm trial done at The Aurum Institute (Tembisa Clinical Research Site, Tembisa, South Africa), with pharmacokinetic visits done at VxPharma (Pretoria, South Africa). Adults (≥18 years) with HIV infection and undetectable viral load (<40 copies per mL) after at least 8 weeks of efavirenz-based or dolutegravir-based regimens were recruited in three consecutive groups, subject to approval by the independent safety monitoring committee. Participants received 50 mg of daily dolutegravir in place of efavirenz for 8 weeks, then began once-weekly rifapentine (900 mg)-isoniazid (900 mg) for 12 weeks. Groups 1A (n=12) and 1B (n=18) had intensive dolutegravir pharmacokinetic sampling at week 8 (before rifapentine-isoniazid), at week 11 (after the third dose of rifapentine)-isoniazid and at week 16 after the eighth dose. Group 2 (n=30) were treated with the same schedule and had sparse dolutegravir pharmacokinetic sampling at weeks 8, 11, and 16. Participants were followed 4 weeks after completion of prophylactic tuberculosis treatment. HIV viral loads were measured at baseline and at weeks 11 and 24. Primary endpoints were adverse events (grade 3 or higher) and dolutegravir population pharmacokinetics, assessed in participants who began rifapentine-isoniazid. This trial was registered at ClinicalTrials.gov, NCT03435146. Between Jan 24, 2018, and Nov 25, 2018, 61 participants were enrolled into three groups; one participant withdrew (from group 1A). 43 (70%) of 60 participants were women and all participants were black African. Median age was 40 years (IQR 35-48), CD4 cell count was 683 cells per μL (447-935), and body-mass index was 28·9 kg/m2 (24·0-32·9). Three grade 3 adverse events occurred; two elevated creatinine and one hypertension. Rifapentine-isoniazid increased dolutegravir clearance by 36% (relative standard error 13%) resulting in a 26% decrease in dolutegravir area under the curve. Overall geometric mean ratio of trough concentrations with versus without rifapentine-isoniazid was 0·53 (90% CI 0·49-0·56) though this ratio varied by day after rifapentine-isoniazid dose. All but one trough value was above the 90% maximal inhibitory concentration for dolutegravir and HIV viral loads were less than 40 copies per mL in all patients. Our results suggest 12 doses of once-weekly rifapentine-isoniazid can be given for tuberculosis prophylaxis to patients with HIV taking dolutegravir-based antiretroviral therapy, without dose adjustments. Further exploration of the pharmacokinetics, safety, and efficacy in children and pharmacodynamics in individuals naive to antiretroviral therapy is needed. UNITAID.


Assessment of trace metal binding kinetics in the resin phase of diffusive gradients in thin films.
The dynamic technique of diffusive gradients in thin films (DGT), that measures metal speciation in situ, has found wide environmental application. Simple interpretation of the metal accumulation in terms of a solution concentration has assumed that trace metals do not penetrate beyond the surface of the binding layer, but penetration, although theoretically discussed has not yet been directly measured. Multiple binding layers were used to enable analysis of different depths of a DGT binding phase (Chelex-100 or iminodiacetate resins). In simple metal solution (no ligand) at pH 7, metal penetration to the back layer was low and similar for all metals. However, at lower pH up to 42% of an individual metal accumulated in the back resin layer. This was most noticeable for Mn at pH 4 and 5, but Cd and Co were also affected at pH 4. These results were consistent with rate limited binding, particularly for Mn. A kinetic model successfully fitted the data and allowed derivation of a binding rate constant and the mean distance that metals penetrate into a resin gel (λ(M)). Only for Mn, Co and Cd were experimentally derived λ(M) values greater than the diameter of a Chelex-100 resin bead. For most situations, then, the penetration into the binding layer is negligible and binding of trace metal ions can be regarded as instantaneous, validating the simple use and interpretation of DGT. For weakly binding metals at low pH the slower binding allows penetration, which may affect the DGT measurement.


Atlantic salmon (Salmo salar L.) as a marine functional source of gamma-tocopherol.
Gamma tocopherol (gT) exhibits beneficial cardiovascular effects partly due to its anti-inflammatory activity. Important sources of gT are vegetable oils. However, little is known to what extent gT can be transferred into marine animal species such as Atlantic salmon by feeding. Therefore, in this study we have investigated the transfer of dietary gT into salmon. To this end, fish were fed a diet supplemented with 170 ppm gT for 16 weeks whereby alpha tocopherol levels were adjusted to 190 ppm in this and the control diet. Feeding gT-rich diets resulted in a three-fold increase in gT concentrations in the liver and fillet compared to non-gT-supplemented controls. Tissue alpha tocopherol levels were not decreased indicating no antagonistic interaction between gamma- and alpha tocopherol in salmon. The concentration of total omega 3 fatty acids slightly increased in response to dietary gT. Furthermore, dietary gT significantly decreased malondialdehyde in the fillet, determined as a biomarker of lipid peroxidation. In the liver of gT fed salmon we observed an overall down-regulation of genes involved in lipid homeostasis. Additionally, gT improved the antioxidant capacity by up-regulating Gpx4a gene expression in the pyloric caeca. We suggest that Atlantic salmon may provide a marine functional source capable of enriching gT for human consumption.


Quantifying bluetongue vertical transmission in French cattle from surveillance data.
Bluetongue is a vector-borne disease of ruminants with economic consequences for the livestock industry. Bluetongue virus serotype 8 (BTV-8) caused a massive outbreak in Europe in 2006/2009 and re-emerged in France in 2015. Given the unprecedented epidemiological features of this serotype in cattle, the importance of secondary routes of transmission was reconsidered and transplacental transmission of BTV-8 was demonstrated in naturally and experimentally infected cattle. Here we used surveillance data from the on-going outbreak to quantify BTV-8 vertical transmission in French cattle. We used RT-PCR pre-export tests collected from June to December 2016 on the French territory and developed a catalytic model to disentangle vertical and vector-borne transmission. A series of in silico experiments validated the ability of our framework to quantify vertical transmission provided sufficient prevalence levels. By applying our model to an area selected accordingly, we estimated a probability of vertical transmission of 56% (55.8%, 95% credible interval 41.7-70.6) in unvaccinated heifers infected late in gestation. The influence of this high probability of vertical transmission on BTV-8 spread and persistence should be further investigated.


Women's experiences of labour and birth when having a termination of pregnancy for fetal abnormality in the second trimester of pregnancy: A qualitative meta-synthesis.
to explore women's experiences of labour and birth in the context of a termination of pregnancy for fetal abnormality in the second trimester of pregnancy. meta-synthesis of 10 qualitative studies which included the experiences of 581 women. Data analysis was informed by van Manen's four lifeworld existentials (lived body, lived space, lived time and lived human relationships) and focused only on women's experiences of their labour and birth when terminating a pregnancy in the second trimester for fetal abnormality. eight themes were generated by the analysis. In my head: a storm of emotion reflected the lived or felt space. In this space women make meaning of the experience. Too late to turn back time and Living and escaping the moment considered lived time; it is lived time that enables a woman to reinterpret who they once were and who they are becoming. The existential concept of lived body represented the women's physical or bodily presence and was reflected in the themes; The language of labour: un-describable torture, The meaning of pain; punishment and protection and Being a mother in the space where birth meets death. Relational or the lived human relationship is reflected in See me… talk to me…hear me… be with me and Sorry baby. These themes describe the women's lived sense of others in the interpersonal space that they share with them. the findings of this meta-synthesis provide insight into how emotionally and physically traumatic a woman's labour and birth experiences can be in this context. Women wanted to spend time with their baby constructing lasting memories that they could hold onto and share. They needed acknowledgment that their baby existed and their loss was tangible and real. What happened within the space of the relationship women shared with care providers was seen as vital, especially when respectful and dignified interaction was experienced. The women's need for human contact and support through their experience was also seen as critical. the need for greater understanding and acknowledgement of the woman's experience by all caregivers is required. Woman-centred and situation sensitive care provision should be prioritised. Further research that acknowledges and prioritises the feminine voice and the lived experience of women is required.


Vertebral bending mechanics and xenarthrous morphology in the nine-banded armadillo (Dasypus novemcinctus).
The vertebral column has evolved to accommodate the broad range of locomotor pressures found across vertebrate lineages. Xenarthran (armadillos, sloths and anteaters) vertebral columns are characterized by xenarthrous articulations, novel intervertebral articulations located in the posterior trunk that are hypothesized to stiffen the vertebral column to facilitate digging. To determine the degree to which xenarthrous articulations impact vertebral movement, we passively measured compliance and range of motion during ventroflexion, dorsiflexion and lateral bending across the thoracolumbar region of the nine-banded armadillo, Dasypus novemcinctus Patterns of bending were compared with changes in vertebral morphology along the column to determine which morphological features best predict intervertebral joint mechanics. We found that compliance was lower in post-diaphragmatic, xenarthrous vertebrae relative to pre-xenarthrous vertebrae in both sagittal and lateral planes of bending. However, we also found that range of motion was higher in this region. These changes in mechanics are correlated with the transition from pre-xenarthrous to xenarthrous vertebrae, as well as with the transition from thoracic to lumbar vertebrae. Our results thus substantiate the hypothesis that xenarthrous articulations stiffen the vertebral column. Additionally, our data suggest that xenarthrous articulations, and their associated enlarged metapophyses, also act to increase the range of motion of the post-diaphragmatic region. We propose that xenarthrous articulations perform the dual role of stiffening the vertebral column and increasing mobility, resulting in passively stable vertebrae that are capable of substantial bending under appropriate loads.


Parotid Gland Nodular Fasciitis: A Clinicopathologic Series of 12 Cases with a Review of 18 Cases from the Literature.
Nodular fasciitis (NF), very uncommon in the parotid gland, is a benign myofibroblastic proliferation that may be mistaken for other neoplastic proliferations. The mass-like clinical presentation and histologic features result in frequent misclassification, resulting in inappropriate clinical management. There are only a few reported cases in the English literature. Cases within the files of the authors' institutions (retrospective) confined to the parotid gland were compared to cases reported in the English literature (Medline 1966-2014). The patients included five females and seven males, aged 11-70 years (mean 45.2 years). All patients presented with a mass lesion, present on average 1.9 months, without a documented history of trauma. The lesions were 0.7-5.2 cm (mean 2.2 cm). Seven patients had fine needle aspiration. The majority of the lesions were circumscribed (n = 9), composed of spindle-shaped to stellate myofibroblasts (MF) arranged in a storiform growth pattern, juxtaposed to hypocellular myxoid tissue-culture-like areas with extravasation of erythrocytes. Dense, keloid-like collagen (n = 7) and occasional giant cells were seen (n = 6). Mitotic figures (without atypical forms) were readily identifiable (mean 4/10 HPFs). By immunohistochemical staining, the MF were reactive with vimentin, actins, and calponin, while the histiocytes were reactive with CD68. All patients had surgical excision. One patient developed local recurrence (12 months later). All were alive and disease free at last follow-up, with a mean 133 months of follow-up. The principle differential diagnoses include fibrosarcoma, fibromatosis, pleomorphic adenoma, myoepithelioma, neurofibroma, schwannoma, solitary fibrous tumor, leiomyoma, fibrous histiocytoma and myxoma. NF of the parotid gland occurs in middle-aged patients who present with a mass (mean 2.2 cm) in the parotid gland of short duration (1.9 months). FNA misinterpretation frequently leads to excision. Separation from myoepithelial and mesenchymal lesions affecting the parotid gland results in appropriate management.


Sunlight-powered kHz rotation of a hemithioindigo-based molecular motor.
Photodriven molecular motors are able to convert light energy into directional motion and hold great promise as miniaturized powering units for future nanomachines. In the current state of the art, considerable efforts have still to be made to increase the efficiency of energy transduction and devise systems that allow operation in ambient and non-damaging conditions with high rates of directional motions. The need for ultraviolet light to induce the motion of virtually all available light-driven motors especially hampers the broad applicability of these systems. We describe here a hemithioindigo-based molecular motor, which is powered exclusively by nondestructive visible light (up to 500 nm) and rotates completely directionally with kHz frequency at 20 °C. This is the fastest directional motion of a synthetic system driven by visible light to date permitting materials and biocompatible irradiation conditions to establish similarly high speeds as natural molecular motors.


Activation of β-catenin signalling by TFF1 loss promotes cell proliferation and gastric tumorigenesis.
In this study, we investigated the role of Trefoil factor 1 (TFF1) in regulating cell proliferation and tumour development through β-catenin signalling using in vivo and in vitro models of gastric tumorigenesis. Tff1-knockout (Tff1-KO) mice, immunohistochemistry, luciferase reporter, qRT-PCR, immunoblot, and phosphatase assays were used to examine the role of TFF1 on β-catenin signalling pathway. Nuclear localisation of β-catenin with transcriptional upregulation of its target genes, c-Myc and Ccnd1, was detected in hyperplastic tissue at an early age of 4-6 weeks and maintained during all stages of gastric tumorigenesis in the Tff1-KO mice. The reconstitution of TFF1 or TFF1 conditioned media significantly inhibited the β-catenin/T-cell factor (TCF) transcription activity in MKN28 gastric cancer cells. In agreement with these results, we detected a reduction in the levels of nuclear β-catenin with downregulation of c-MYC and CCND1 mRNA. Analysis of signalling molecules upstream of β-catenin revealed a decrease in phosphorylated glycogen synthase kinase 3β (p-GSK3β) (Ser9) and p-AKT (Ser473) protein levels following the reconstitution of TFF1 expression; this was consistent with the increase of p-β-catenin (Ser33/37/Thr41) and decrease of p-β-catenin (Ser552). This TFF1-induced reduction in phosphorylation of GSK3β, and AKT was dependent on protein phosphatase 2A (PP2A) activity. The treatment with okadaic acid or knockdown of PP2A abrogated these effects. Consistent with the mouse data, we observed loss of TFF1 and an increase in nuclear localisation of β-catenin in stages of human gastric tumorigenesis. Our data indicate that loss of TFF1 promotes β-catenin activation and gastric tumorigenesis through regulation of PP2A, a major regulator of AKT-GSK3β signalling.


Preoperative embolization in surgical treatment of spinal metastases: single-blind, randomized controlled clinical trial of efficacy in decreasing intraoperative blood loss.
To assess whether preoperative transcatheter arterial embolization of spinal metastases reduces blood loss, the need for transfusion with allogeneic red blood cells (RBCs), and surgery time in the surgical treatment of patients with symptomatic metastatic spinal cord compression. This single-blind, randomized (1:1), controlled, parallel-group, single-center trial was approved by the Danish National Committee on Biomedical Research Ethics and was conducted from May 2011-March 2013. Participants (N = 45) were scheduled for decompression and posterior thoracic/lumbar spinal instrumentation and randomly assigned to either preoperative embolization (n = 23) or a control group (n = 22). The primary outcome was intraoperative blood loss. Secondary outcomes were perioperative blood loss, allogeneic RBC transfusion, and surgery time. Analyses were performed by intention-to-treat. The intention-to-treat analysis included 45 patients. Mean intraoperative blood loss did not differ significantly (P = .270) between the embolization group (618 mL [SD, 282 mL]) and the control group (735 mL [SD, 415 mL]). There was also no significant difference in allogeneic RBC transfusion (P = .243). Surgery time was significantly shorter in the embolization group (P = .031): median 90 minutes (range, 54-252 min) versus 124 minutes (range, 80-183 min). The subanalysis of hypervascular metastases revealed a significant (P = .041) reduction in blood loss in the embolization group: 645 mL (SD, 289 mL) versus 902 mL (SD, 416 mL). Preoperative embolization in patients with symptomatic spinal metastasis independent of primary tumor diagnosis did not reduce intraoperative blood loss and allogeneic RBC transfusion significantly but did reduce the surgery time. A small reduction of intraoperative blood loss was shown in hypervascular metastases.


Using multispecies occupancy models to improve the characterization and understanding of metacommunity structure.
Two of the most prominent frameworks to develop in ecology over the past decade are metacommunity ecology, which seeks to characterize multispecies distributions across space, and occupancy modeling, which corrects for imperfect detection in an effort to better understand species occurrence patterns. Although their goals are complementary, metacommunity theory and statistical occupancy modeling methods have developed independently. For instance, the elements of metacommunity structure (EMS) framework uses species occurrence data to classify metacommunity structure and link it to underlying environmental gradients. While the efficacy of this approach relies on the quality of the data, few studies have considered how imperfect detection, which is widespread in ecological surveys and the major focus of occupancy modeling, affects the outcome. We introduce a framework that integrates multispecies occupancy models with the current EMS framework, detection error-corrected EMS (DECEMS). This method offers two distinct advantages. First, DECEMS reduces bias in characterizing metacommunity structure by using repeated surveys and occupancy models to disentangle species-specific occupancy and detection probabilities, ultimately bringing metacommunity structure classification into a more probabilistic framework. Second, occupancy modeling allows estimation of species-specific responses to environmental covariates, which will increase our ability to link species-level effects to metacommunity-wide patterns. After reviewing the EMS framework, we introduce a simple multispecies occupancy model and show how DECEMS can work in practice, highlighting that detection error often causes EMS to assign incorrect structures. To emphasize the broader applicability of this approach, we further illustrate that DECEMS can reduce the rate of structure misclassification by more than 20% in some cases, even proving useful when detection error rates are quite low (-10%). Integrating occupancy models and the EMS framework will lead to more accurate descriptions of metacommunity structure and to a greater understanding of the mechanisms by which different structures arise.


Cu boosting the collaborative effect of Ni and H+ in alloyed NiCu/saponite catalysts for hydrogenolysis of glycidol.
The effect of copper on various acid saponite supported Ni-Cu bimetallic catalysts, prepared with different Ni : Cu ratios, was studied for the liquid phase hydrogenolysis of glycidol on a batch reactor at 393 and 453 K. Characterization of the catalysts showed that Ni and Cu are in close contact as the XRD measurements evidenced the formation of an alloy. H2 chemisorption results revealed that the measured metallic area progressively decreased with an increase in the wt% of copper. In the presence of high metal activity (higher Ni wt%), the formation of 1,2-propanediol (1,2-PD) outweighed, while acid activity led to the formation of dimerization and oligomerization products. The addition of Cu and the increase of the reaction temperature decreased the diol formation but boosted the 1,3-PD/1,2-PD ratio. This could be explained by an improvement of the collaborative effect between the metal Ni and the H+ of the saponite. Therefore, the presence of an appropriate amount of Cu allowed the control of the hydrogenation capacity of Ni and enhanced the collaborative effect of Ni and H+ favouring the formation of 1,3-propanediol with respect to 1,2-propanediol.


Indications and results of cordectomy for early glottic carcinoma.
Select early glottic carcinomas in patients with normal vocal cord mobility can be successfully treated with cordectomy, performed either endoscopically or by means of an open operation (laryngofissure-cordectomy). From 1976 to 1986, 95 patients underwent cordectomy at our institution. There were 88 men and 7 women, with an age range at the time of surgery of 27 to 84 years. A laryngofissure was used to perform a cordectomy in 61 patients and an endoscopic approach was used in 34. The median number of days in the hospital for the endoscopic group was 2, and for the laryngofissure group it was 6. The Kaplan-Meier product-limit method showed the probability of no recurrence in the first 3 years after an endoscopic cordectomy was 0.9099, and in the laryngofissure-cordectomy group it was 0.9502. To date, there have been four recurrences (11.8%) in the endoscopic group and three (4.9%) in the laryngofissure group. Only one patient with recurrent tumor is known to have died of the cancer. Laryngectomy was necessary for ultimate control in three patients. The Kaplan-Meier survival curve for the 95 patients was the same as that for a sex- and age-matched control group. Cordectomy is an effective treatment option that results in minimal morbidity for select early glottic carcinomas.


Carnitine-dependent metabolic activities in Saccharomyces cerevisiae: three carnitine acetyltransferases are essential in a carnitine-dependent strain.
L-carnitine is required for the transfer of activated acyl-groups across intracellular membranes in eukaryotic organisms. In Saccharomyces cerevisiae, peroxisomal membranes are impermeable to acetyl-CoA, which is produced in the peroxisome when cells are grown on fatty acids as carbon source. In a reversible reaction catalysed by carnitine acetyltransferases (CATs), activated acetyl groups are transferred to carnitine to form acetylcarnitine which can be shuttled across membranes. Here we describe a mutant selection strategy that specifically selects for mutants affected in carnitine-dependent metabolic activities. Complementation of three of these mutants resulted in the cloning of three CAT encoding genes: CAT2, coding for the carnitine acetyltransferase associated with the peroxisomes and the mitochondria; YAT1, coding for the carnitine acetyltransferase, which is presumably associated with the outer mitochondrial membrane, and YER024w (YAT2), which encodes a third, previously unidentified carnitine acetyltransferase. The data also show that (a) L-carnitine and all three CATs are essential for growth on non-fermentable carbon sources in a strain with a disrupted CIT2 gene; (b) Yat2p contributes significantly to total CAT activity when cells are grown on ethanol; and that (c) the carnitine-dependent transfer of activated acetyl groups plays a more important role in cellular processes than previously realised.


Social withdrawal in childhood.
Socially withdrawn children frequently refrain from social activities in the presence of peers. The lack of social interaction in childhood may result from a variety of causes, including social fear and anxiety or a preference for solitude. From early childhood through to adolescence, socially withdrawn children are concurrently and predictively at risk for a wide range of negative adjustment outcomes, including socio-emotional difficulties (e.g., anxiety, low self-esteem, depressive symptoms, and internalizing problems), peer difficulties (e.g., rejection, victimization, poor friendship quality), and school difficulties (e.g., poor-quality teacher-child relationships, academic difficulties, school avoidance). The goals of the current review are to (a) provide some definitional, theoretical, and methodological clarity to the complex array of terms and constructs previously employed in the study of social withdrawal; (b) examine the predictors, correlates, and consequences of child and early-adolescent social withdrawal; and (c) present a developmental framework describing pathways to and from social withdrawal in childhood.


Vitamin D Levels in Children with Adenotonsillar Hypertrophy and Otitis Media with Effusion.
Vitamin D has been suggested to play a considerable role in the function of the immune system in various infectious, inflammatory, and autoimmune conditions. Otitis media with effusion (OME), defined as the presence of non-purulent fluid within the middle ear without signs or symptoms of suppurative otitis media, has a number of inflammatory predisposing factors. This study was designed to explore the association between vitamin D deficiency and OME. In this cross-sectional study, 74 children aged 2-7 years with an obstructive indication for adenotonsillectomy were included. Patients were divided into two groups based on the need for ventilation tube insertion for OME. Thirty-two children were enrolled in the OME group and 42 in the control group. The mean vitamin D level was compared between the two groups. Mean vitamin D concentration in all patients was 11.96±5.85 ng/ml (9.79±4.36 ng/ml in the OME group and 13.61±6.33 ng/ml in the control group; P=0.003). There was also a significant difference in levels of vitamin D in patients referred in winter (9.0±2.94 ng/ml) compared with the summer (19.85±4.21 ng/ml; P=0.001). Data analyzed based on the season in which the patients were referred showed no significant difference between the OME and the control group. Although our results showed lower serum levels of vitamin D in OME patients, the difference was not significant when seasons were taken into consideration. Therefore, the season is an important confounding factor in any research related to vitamin D due to the effect of sun-induced vitamin D.


Outcome of endoscopic minor papillotomy in patients with symptomatic pancreas divisum.
Pancreas divisum has been associated with recurrent acute pancreatitis, chronic abdominal pain without elevated pancreatic enzymes, and chronic pancreatitis. Prior studies suggest that endoscopic minor papillotomy benefits certain symptomatic pancreas divisum patients. However, the data are quite limited and there is a lack of long-term follow-up. To describe a retrospective study of endoscopic minor papillotomy for pancreas divisum. Eighty-nine adult patients who underwent endoscopic minor papillotomy at our referral center were included in the study. Median follow-up was 29 months. We conducted a telephone survey. Fifty-three patients were available for the telephone survey. Thirty-two patients (60.4%) reported immediate improvement: however, symptoms recurred in 17 (53.1% of the immediate responders). Repeat endoscopic interventions were performed in 8 patients, with long-term improvement in two. Overall long-term improvement was achieved in 17 patients (32.1%). Results of minor papillotomy were more favorable for patients with recurrent, well-defined bouts of pancreatitis (immediate improvement: P=0.036; long-term improvement: P=0.064) compared to those with pancreatitis who reported continuous pain and those without clinical evidence of pancreatitis (immediate improvement: 73.3%, 42.9% and 44.4%, respectively; long-term improvement: 43.3%, 21.4%, and 11.1%, respectively). The long-term benefit from endoscopic minor papillotomy using strict criteria is poorer than suggested from previous studies. However, pancreas divisum patients with well-defined bouts of pancreatitis are more likely to benefit from endoscopic minor papillotomy than those without symptom-free intervals between "attacks" and those with pain that is not associated with elevated pancreatic enzymes.


De novo nonsense mutations in ASXL1 cause Bohring-Opitz syndrome.
Bohring-Opitz syndrome is characterized by severe intellectual disability, distinctive facial features and multiple congenital malformations. We sequenced the exomes of three individuals with Bohring-Opitz syndrome and in each identified heterozygous de novo nonsense mutations in ASXL1, which is required for maintenance of both activation and silencing of Hox genes. In total, 7 out of 13 subjects with a Bohring-Opitz phenotype had de novo ASXL1 mutations, suggesting that the syndrome is genetically heterogeneous.


Integrating multi-scale data to create a virtual physiological mouse heart.
While the virtual physiological human (VPH) project has made great advances in human modelling, many of the tools and insights developed as part of this initiative are also applicable for facilitating mechanistic understanding of the physiology of a range of other species. This process, in turn, has the potential to provide human relevant insights via a different scientific path. Specifically, the increasing use of mice in experimental research, not yet fully complemented by a similar increase in computational modelling, is currently missing an important opportunity for using and interpreting this growing body of experimental data to improve our understanding of cardiac function. This overview describes our work to address this issue by creating a virtual physiological mouse model of the heart. We describe the similarities between human- and mouse-focused modelling, including the reuse of VPH tools, and the development of methods for investigating parameter sensitivity that are applicable across species. We show how previous results using this approach have already provided important biological insights, and how these can also be used to advance VPH heart models. Finally, we show an example application of this approach to test competing multi-scale hypotheses by investigating variations in length-dependent properties of cardiac muscle.


Epithelial cell polarity affects susceptibility to Pseudomonas aeruginosa invasion and cytotoxicity.
Intact tissues are relatively resistant to Pseudomonas aeruginosa-induced disease, and injury predisposes tissue to infection. Intact epithelia contain polarized cells that have distinct apical and basolateral membranes with unique lipids and proteins. In this study, the role of cell polarity in epithelial cell susceptibility to P. aeruginosa virulence mechanisms was tested. Madin-Darby canine kidney (MDCK) cells, human corneal epithelial cells, and primary cultures of two different types of airway epithelial cells were grown on Transwell filters or in plastic tissue culture wells. P. aeruginosa invasion of cells was quantified by gentamicin survival assays with two isolates that invade epithelial cells (6294 and PAO1). Cytotoxic activity was assessed by trypan blue exclusion assays with two cytotoxic strains (6206 and PA103). Basolateral surfaces of cells were exposed by one of two methods: EGTA pretreatment of epithelial cells or growth of cells in low-calcium medium. Both methods of exposing basolateral membranes increased epithelial cell susceptibility to P. aeruginosa invasion and cytotoxicity. Migrating cells were also found to be more susceptible to P. aeruginosa invasion than confluent monolayers that had established membrane polarity. Monolayers of MDCK cells that had been selected for resistance to killing by concanavalin A were resistant to both cytotoxicity and invasion by P. aeruginosa because they were more efficiently polarized for their susceptibility to P. aeruginosa virulence factors than regular MDCK cells and not because they were defective in glycosylation. These results suggest that there are factors on the basolateral surfaces of epithelial cells that promote interaction with P. aeruginosa or that there are inhibitory factors on the apical cell surface. Thus, cell polarity of intact epithelia is likely to contribute to defense against P. aeruginosa infection.


[The history of developing anticancer Drugs and their evaluation guidelines in Japan].
The cancer therapies currently available do not yet offer fully satisfactory treatments, even in 21st century, and efforts and progress are being made daily in the area of drug development. Anticancer drugs, which play the leading role in cancer therapy, are being developed dynamically around the world, and Japan is not an exception. Looking back on the history of developing anticancer drugs, cytotoxic drugs were the mainstream of drug development until the end of the 20th century. In the 21st century, they have been replaced by molecularly targeted drugs, and thus the development of cytotoxic drugs has been declining rapidly. There were various approaches to the development of anticancer drugs and clinical trial endpoints until the 1980s. In 1991, the "Guidelines for Clinical Evaluation Methods of Anti-Cancer Drugs in Japan" was issued. From 2000 onwards, there was vigorous discussion on the clinical trial endpoints of anticancer drugs in the United States. In conjunction with this discussion, the "Guidelines for Clinical Evaluation Methods of Anti-Cancer Drugs in Japan" was revised in 2005. The revised guidelines required survival data at the time of filing a new drug application (NDA) as a general rule. Around 2005, a bridging strategy was promoted as the "International Conference on Harmonization E5" was promulgated among Japan, the U.S. and EU, resulting in an outflow of clinical trials to overseas, with more non-Japanese survival data generated outside of Japan used for NDAs than Japanese data. Subsequently, the "Guideline for Basic Principles on Global Clinical Trials" was issued in 2007, which promoted the change in the mainstream approach from a bridging strategy to a pivotal, global study involving Japan. Thus, an era of full-fledged globalization in clinical trials began. We believe Japan will need systems to enhance the motivation for anticancer drug development, such as an expedited program or pediatric program, from now on. We hope that the enhancement of these systems will contribute to shortening the period required for approving an anticancer drug and reducing developmental costs. Furthermore, we expect Japan to be creating breakthrough anticancer drugs in the near future.


Cytokine patterns and viral load in lymph nodes during the early stages of SIV infection.
To elucidate the initial pathogenic events in lymphoid organs, the major reservoir of virus in HIV infection, follow-up of viral load and cytokine mRNA production was performed in the rhesus macaque SIV mac251 model. In the first experiment, lymph nodes (LN) obtained from animals sacrificed at early time points following intravenous (i.v.) inoculation with a high viral dose, showed that the production of cytokine mRNA in LN (IFN gamma, IL1 beta, IL2, IL4, IL12p40, IL6, IL10, TNF alpha and TNF beta) was correlated with viral replication and persisted during the two months post-inoculation (p.i.). In the second experiment, LN were sequentially analysed in two groups of animals receiving i.v. a high or a low viral dose. The initial higher local production of IFN gamma mRNA in LN was associated with weak viraemia, the capacity of the host to decrease productive infection in LN measured at one and two months p.i., and slow disease progression.