The burden of chronic spontaneous urticaria is substantial: Real-world evidence from ASSURE-CSU.
Chronic spontaneous urticaria (CSU) can be debilitating, difficult to treat, and frustrating for patients and physicians. Real-world evidence for the burden of CSU is limited. The objective of this study was to document disease duration, treatment history, and disease activity, as well as impact on health-related quality of life (HRQoL) and work among patients with inadequately controlled CSU, and to describe its humanistic, societal, and economic burden. This international observational study assessed a cohort of 673 adult patients with CSU whose symptoms persisted for ≥12 months despite treatment. Demographics, disease characteristics, and healthcare resource use in the previous 12 months were collected from medical records. Patient-reported data on urticaria and angioedema symptoms, HRQoL, and work productivity and activity impairment were collected from a survey and a diary. Almost 50% of patients had moderate-to-severe disease activity as reported by Urticaria Activity Score. Mean (SD) Dermatology Life Quality Index and Chronic Urticaria Quality of Life Questionnaire scores were 9.1 (6.62) and 33.6 (20.99), respectively. Chronic spontaneous urticaria markedly interfered with sleep and daily activities. Angioedema in the previous 12 months was reported by 66% of enrolled patients and significantly affected HRQoL. More than 20% of patients reported ≥1 hour per week of missed work; productivity impairment was 27%. These effects increased with increasing disease activity. Significant healthcare resources and costs were incurred to treat CSU. Chronic spontaneous urticaria has considerable humanistic and economic impacts. Patients with greater disease activity and with angioedema experience greater HRQoL impairments.


m6A modification of non-coding RNA and the control of mammalian gene expression.
The biology of non-coding RNA (ncRNA) and the regulation of mammalian gene expression is a rapidly expanding field. In this review, we consider how recent advances in technology, enabling the precise mapping of modifications to RNA transcripts, has provided new opportunities to dissect post-transcriptional gene regulation. With this has come the realisation that in the absence of translation, the modification of ncRNAs may play a fundamental role in their regulation, protein interactome and subsequent downstream effector functions. We focus upon modification of RNA by N6-methyladenosine (m6A); its readers, writers and erasers, before considering the differing role of m6A modified lncRNAs MALAT1 and Xist. This article is part of a Special Issue entitled: mRNA modifications in gene expression control edited by Dr. Soller Matthias and Dr. Fray Rupert.


[Evaluation of a cervical cancer control program in a local health system in Southeast Brazil].
This study considers the possibility of expanding the roles of the local health care system to respond to demands by the population. Focusing on the problem of cervical cancer, the study evaluated the results obtained by a preventive program conducted in a municipal health system with the objective of evaluating its case-resolving capacity and patient compliance. Variables were related to diagnosis and follow-up of 465 women enrolled in the program from 1987 to 1994. For purposes of comparison, the study used the results from a specialized regional referral service during the same period. The program's case-resolving capacity was similar to that of the referral service; in addition, on average the cervical lesions were less severe and the treatment drop-out rate was lower among patients from the local health care system, reflecting better follow-up compliance. It is important to train local health personnel in order to improve care, including treatment for more complex diseases.


Interplay of proton and electron transfer to determine concerted behavior in the proton-coupled electron transfer of glutathione oxidation.
Glutathione (GSH), whose thiol group dictates its redox chemistry, is oxidized to the thiyl radical (GS˙), which rapidly dimerizes to GSSG. Previously, we found that the oxidation rate of GSH by IrCl62- depends on the base (B) concentration and the pKa of its conjugate acid BH+, so that collateral to a stepwise mechanism, the concerted pathway GSH + IrCl62- + B = GS˙ + IrCl63- + BH+ was proposed as the rate determining step. Herein, this investigation is extended to include oxidant-base pairs that render exothermic and endothermic conditions of ΔG°' for electron transfer (ET) and proton transfer (PT). Experiments were conducted by the reaction of GSH with an electrogenerated oxidant M+ and using digital simulations to infer the mechanism. Data analysis shows that despite parallel mechanisms, the concerted one seems to predominate for the oxidant-base pair that renders the most isoenergetic coupled state, whereby a PT with is capable of producing an ET with , as a result of the Nernstian shift of with pKa. In contrast, the stepwise PT-ET appears to dominate when GS- grows in stability as becomes more negative. Understanding the interplay between ET and PT will help in the design of catalysts for energy harvesting processes that rely on proton-coupled electron transfer.


Association of coffee intake with reduced incidence of liver cancer and death from chronic liver disease in the US multiethnic cohort.
Coffee consumption has been proposed to reduce risk for hepatocellular carcinoma (HCC) and chronic liver disease (CLD), but few data are available from prospective, US multiethnic populations. We evaluated the association of coffee intake with HCC and CLD in 162,022 African Americans, Native Hawaiians, Japanese Americans, Latinos, and whites in the US Multiethnic Cohort (MEC). We collected data from the MEC, a population-based prospective cohort study of >215,000 men and women from Hawaii and California, assembled in 1993-1996. Participants reported coffee consumption and other dietary and lifestyle factors when they joined the study. During an 18-year follow-up period, there were 451 incident cases of HCC and 654 deaths from CLD. Hazard rate ratios (RRs) and 95% confidence intervals (CIs) were calculated using Cox regression, adjusting for known HCC risk factors. High levels of coffee consumption were associated with reduced risk of incident HCC and CLD mortality (Ptrend ≤ .0002). Compared with non-coffee drinkers, those who drank 2-3 cups per day had a 38% reduction in risk for HCC (RR = 0.62; 95% CI: 0.46-0.84); those who drank ≥4 cups per day had a 41% reduction in HCC risk (RR = 0.59; 95% CI: 0.35-0.99). Compared with non-coffee drinkers, participants who consumed 2-3 cups coffee per day had a 46% reduction in risk of death from CLD (RR = 0.54; 95% CI: 0.42-0.69) and those who drank ≥4 cups per day had a 71% reduction (RR = 0.29; 95% CI: 0.17-0.50). The inverse associations were similar regardless of the participants' ethnicity, sex, body mass index, smoking status, alcohol intake, or diabetes status. Increased coffee consumption reduces the risk of HCC and CLD in multiethnic US populations.


Combined OCT distance and FBG force sensing cannulation needle for retinal vein cannulation: in vivo animal validation.
Retinal vein cannulation is an experimental procedure during which a clot-dissolving drug is injected into an obstructed retinal vein. However, due to the fragility and minute size of retinal veins, such procedure is considered too risky to perform manually. With the aid of surgical robots, key limiting factors such as: unwanted eye rotations, hand tremor and instrument immobilization can be tackled. However, local instrument anatomy distance and force estimation remain unresolved issues. A reliable, real-time local interaction estimation between instrument tip and the retina could be a solution. This paper reports on the development of a combined force and distance sensing cannulation needle, and its experimental validation during in vivo animal trials. Two prototypes are reported, relying on force and distance measurements based on FBG and OCT A-scan fibres, respectively. Both instruments provide an 80 [Formula: see text] needle tip and have outer shaft diameters of 0.6 and 2.3 mm, respectively. Both prototypes were characterized and experimentally validated ex vivo. Then, paired with a previously developed surgical robot, in vivo experimental validation was performed. The first prototype successfully demonstrated the feasibility of using a combined force and distance sensing instrument in an in vivo setting. The results demonstrate the feasibility of deploying a combined sensing instrument in an in vivo setting. The performed study provides a foundation for further work on real-time local modelling of the surgical scene. This paper provides initial insights; however, additional processing remains necessary.


Recognition of T*G mismatched base pairs in DNA by stacked imidazole-containing polyamides: surface plasmon resonance and circular dichroism studies.
An imidazole-containing polyamide trimer, f-ImImIm, where f is a formamido group, was recently found using NMR methods to recognize T*G mismatched base pairs. In order to characterize in detail the T*G recognition affinity and specificity of imidazole-containing polyamides, f-ImIm, f-ImImIm and f-PyImIm were synthesized. The kinetics and thermodynamics for the polyamides binding to Watson-Crick and mismatched (containing one or two T*G, A*G or G*G mismatched base pairs) hairpin oligonucleotides were determined by surface plasmon resonance and circular dichroism (CD) methods. f-ImImIm binds significantly more strongly to the T*G mismatch-containing oligonucleotides than to the sequences with other mismatched or with Watson-Crick base pairs. Compared with the Watson-Crick CCGG sequence, f-ImImIm associates more slowly with DNAs containing T*G mismatches in place of one or two C*G base pairs and, more importantly, the dissociation rate from the T*G oligonucleotides is very slow (small k(d)). These results clearly demonstrate the binding selectivity and enhanced affinity of side-by-side imidazole/imidazole pairings for T*G mismatches and show that the affinity and specificity increase arise from much lower k(d) values with the T*G mismatched duplexes. CD titration studies of f-ImImIm complexes with T*G mismatched sequences produce strong induced bands at approximately 330 nm with clear isodichroic points, in support of a single minor groove complex. CD DNA bands suggest that the complexes remain in the B conformation.


Quantification of elemental and organic carbon in atmospheric particulate matter using color space sensing-hue, saturation, and value (HSV) coordinates.
A fast and cost effective application of color sensing was used to quantify color coordinates of atmospheric particulate matter collected on filters to quantify elemental and organic carbon (EC/OC) loading. This is a unique and novel approach for estimating OC composition. The method used a colorimeter and digital photography to obtain XYZ color space values and mathematically transformed them to HSV cylindrical-coordinates; a quantification method was applied to estimate the NIOSH and IMPROVE (TOR) EC/OC loadings from a set of globally diverse PM samples. When applied to 315 samples collected at three US EPA Chemical Speciation Network (CSN) sampling sites, the HSV model proved to be a robust method for EC measurement with an R(2)=0.917 for predicted versus measured loading results and a CV(RMSE)=16.1%. The OC quantified from the same sample filters had an R(2)=0.671 and a CV(RMSE)=24.8% between the predicted and measured results. The method was applied to NIOSH EC/OC results from a set of samples from rural China, Bagdad, and the San Joaquin Valley, CA, and the EC and OC CV(RMSE) were 30.8% and 49.3%, respectively. Additionally, the method was applied to samples with color quantified by a digital photographic image (DPI) with EC results showing good agreement with a CV(RMSE) of 22.6%. OC concentrations were not captured as accurately with the DPI method, with a CV(RMSE) of 77.5%. The method's low analytical cost makes it a valuable tool for estimating EC/OC exposure in developing regions and for large scale monitoring campaigns.


Peri-implant infections of oral biofilm etiology.
Biofilms are complex microbial communities that grow on various surfaces in nature. The oral micobiota tend to form polymicrobial biofilms, particularly on the hard mineralized surfaces of teeth, which may impact on oral health and disease. They can cause inflammation of the adjacent tooth-supporting (periodontal) tissues, leading to destructive periodontal disease and tooth loss. The emergence of osseointegrated dental implants as a restorative treatment option for replacing missing teeth has also brought along new artificial surfaces within the oral cavity, on which oral bacteria can form biofilms. As in the case of natural teeth, biofilms on implant surfaces may also trigger infection and cause inflammatory destruction of the peri-implant tissue (i.e. peri-implantitis). While there are strong similarities in the composition of the mixed microbial flora between periodontal and peri-implant infections, there are also a few distinctive differences. The immunological events underlying the pathogenesis of peri-implant infections are qualitatively similar, yet more extensive, compared to periodontal infections, resulting in a faster progression of tissue destruction. This chapter summarizes the current knowledge on the microbiology and immunology of peri-implant infections, including findings from the peri-implant crevicular fluid, the inflammatory exudate of the peri-implant tissue. Moreover, it discusses the diagnosis and current approaches for the treatment of oral infections.


Biochip-based study of unidirectional mitochondrial transfer from stem cells to myocytes via tunneling nanotubes.
Tunneling nanotubes (TNTs) are small membranous tubes of 50-1000 nm diameter observed to connect cells in culture. Transfer of subcellular organelles through TNTs was observed in vitro and in vivo, but the formation and significance of these structures is not well understood. A polydimethylsiloxane biochip-based coculture model was devised to constrain TNT orientation and explore both TNT-formation and TNT-mediated mitochondrial transfer. Two parallel microfluidic channels connected by an array of smaller microchannels enabled localization of stem cell and cardiomyocyte populations while allowing connections to form between them. Stem cells and cardiomyocytes were deposited in their respective microfluidic channels, and stem cell-cardiomyocyte pairs were formed via the microchannels. Formation of TNTs and transfer of stained mitochondria through TNTs was observed by 24 h real-time video recording. The data show that stem cells are 7.7 times more likely to initiate contact by initial extension of filopodia. By 24 h, 67% of nanotube connections through the microchannels are composed of cardiomyocyte membrane. Filopodial extension and retraction by stem cells draws an extension of TNTs from cardiomyocytes. MitoTracker staining shows that unidirectional transfer of mitochondria between stem cell-cardiomyocyte pairs invariably originates from stem cells. Control experiments with cardiac fibroblasts and cardiomyocytes show little nanotube formation between homotypic or mixed cell pairs and no mitochondrial transfer. These data identify a novel biological process, unidirectional mitochondrial transfer, mediated by heterotypic TNT connections. This suggests that the enhancement of cardiomyocyte function seen after stem-cell injection may be due to a bioenergetic stimulus provided by mitochondrial transfer.


Comparative analysis of Faecalibacterium prausnitzii genomes shows a high level of genome plasticity and warrants separation into new species-level taxa.
Faecalibacterium prausnitzii is a ubiquitous member of the human gut microbiome, constituting up to 15% of the total bacteria in the human gut. Substantial evidence connects decreased levels of F. prausnitzii with the onset and progression of certain forms of inflammatory bowel disease, which has been attributed to its anti-inflammatory potential. Two phylogroups of F. prausnitzii have been identified, with a decrease in phylogroup I being a more sensitive marker of intestinal inflammation. Much of the genomic and physiological data available to date was collected using phylogroup II strains. Little analysis of F. prausnitzii genomes has been performed so far and genetic differences between phylogroups I and II are poorly understood. In this study we sequenced 11 additional F. prausnitzii genomes and performed comparative genomics to investigate intraspecies diversity, functional gene complement and the mobilome of 31 high-quality draft and complete genomes. We reveal a very low level of average nucleotide identity among F. prausnitzii genomes and a high level of genome plasticity. Two genomogroups can be separated based on differences in functional gene complement, albeit that this division does not fully agree with separation based on conserved gene phylogeny, highlighting the importance of horizontal gene transfer in shaping F. prausnitzii genomes. The difference between the two genomogroups is mainly in the complement of genes associated with catabolism of carbohydrates (such as a predicted sialidase gene in genomogroup I) and amino acids, as well as defense mechanisms. Based on the combination of ANI of genomic sequences, phylogenetic analysis of core proteomes and functional differences we propose to separate the species F. prausnitzii into two new species level taxa: F. prausnitzii sensu stricto (neotype strain A2-165T = DSM 17677T = JCM 31915T) and F. moorei sp. nov. (type strain ATCC 27768T = NCIMB 13872T).


Reasoning like a doctor or like a nurse? A systematic integrative review.
When physicians and nurses are looking at the same patient, they may not see the same picture. If assuming that the clinical reasoning of both professions is alike and ignoring possible differences, aspects essential for care can be overlooked. Understanding the multifaceted concept of clinical reasoning of both professions may provide insight into the nature and purpose of their practices and benefit patient care, education and research. We aimed to identify, compare and contrast the documented features of clinical reasoning of physicians and nurses through the lens of layered analysis and to conduct a simultaneous concept analysis. The protocol of this systematic integrative review was published doi: 10.1136/bmjopen-2021-049862. A comprehensive search was performed in four databases (PubMed, CINAHL, Psychinfo, and Web of Science) from 30th March 2020 to 27th May 2020. A total of 69 Empirical and theoretical journal articles about clinical reasoning of practitioners were included: 27 nursing, 37 medical, and five combining both perspectives. Two reviewers screened the identified papers for eligibility and assessed the quality of the methodologically diverse articles. We used an onion model, based on three layers: Philosophy, Principles, and Techniques to extract and organize the data. Commonalities and differences were identified on professional paradigms, theories, intentions, content, antecedents, attributes, outcomes, and contextual factors. The detected philosophical differences were located on a care-cure and subjective-objective continuum. We observed four principle contrasts: a broad or narrow focus, consideration of the patient as such or of the patient and his relatives, hypotheses to explain or to understand, and argumentation based on causality or association. In the technical layer a difference in the professional concepts of diagnosis and the degree of patient involvement in the reasoning process were perceived. Clinical reasoning can be analysed by breaking it down into layers, and the onion model resulted in detailed features. Subsequently insight was obtained in the differences between nursing and medical reasoning. The origin of these differences is in the philosophical layer (professional paradigms, intentions). This review can be used as a first step toward gaining a better understanding and collaboration in patient care, education and research across the nursing and medical professions.


Oncogene expression in autoimmune and normal peripheral blood mononuclear cells.
PBMC from patients with autoimmune diseases and from normal controls were studied for the expression of several cellular oncogenes. Gene expression was assessed by Northern blot analysis of poly(A)+ RNA obtained from leukapheresis samples. Patients with SLE expressed significantly more c-myc protooncogene RNA than did normal controls. Increased expression of the N-ras protooncogene was found in that subset of patients whose autoimmune disease was very active. Cells from individuals with SLE, but not from those with other autoimmune illnesses, showed significantly decreased levels of the c-myb and c-fos protooncogenes. To examine the implications of these findings, B and T cells were purified from apheresis samples donated by normal volunteers. When mitogen was used to activate the B cells in vitro, their pattern of protooncogene expression changed to resemble that found in freshly isolated cells from lupus patients. These results suggest that the differences detected in the expression of protooncogenes by patients with SLE may be due to the abnormal activation of their B cells in vivo. The pattern of protooncogene expression found in patients with other autoimmune illnesses is consistent with the activation of additional cell types in those diseases.


Pre-pregnancy body mass index and breastfeeding initiation, early cessation and longevity: evidence from the first wave of the UK Millennium Cohort Study.
International evidence indicates relationships between pre-pregnancy body mass index (BMI) and breastfeeding behaviours. This study aims to assess associations between key points in the breastfeeding trajectory (initiation, early cessation and longevity) and pre-pregnancy BMI in a recent, nationally representative British cohort. It also aims to explore in the British context potential moderation by mothers' ethnic group. The sample comprises 17 113 mothers from the UK Millennium Cohort Study who have information on pre-pregnancy BMI. Associations between pre-pregnancy BMI categories and breastfeeding initiation, early cessation and longevity are tested using logistic regression. Directed acyclic graphics identify appropriate minimal adjustment to block biasing pathways and classify total and direct effects. After adjusting for confounders, there are large differences in breastfeeding early cessation and longevity by pre-pregnancy BMI group. Differences in propensity to initiation are negligible. Having begun breastfeeding, overweight and obese mothers are more likely to cease in the first week and less likely to continue past 4 months. Observed potential mediators within pregnancy and delivery provide little explanation for relationships. Evidence for moderation by ethnicity is scant. The causal mechanisms underlying relationships between pre-pregnancy overweight, obesity, and breastfeeding behaviours require further research. However, this study suggests pre-pregnancy BMI as one predictive measure for targeting support to women less likely to establish breastfeeding in the early days, and to continue beyond 4 months. The nature of support should carefully be considered and developed, with mind to both intended and potential unintended consequences of intervention given the need for additional investigation into the causes of associations.


Bortezomib inhibits PKR-like endoplasmic reticulum (ER) kinase and induces apoptosis via ER stress in human pancreatic cancer cells.
Bortezomib (Velcade, formerly known as PS-341) is a boronic acid dipeptide derivative that is a selective and potent inhibitor of the proteasome. We hypothesized that proteasome inhibition would lead to an accumulation of misfolded proteins in the cell resulting in endoplasmic reticulum (ER) stress. The ability of bortezomib to induce ER stress and the unfolded protein response was investigated in a human pancreatic cancer cell line, L3.6pl. Bortezomib increased expression of ER stress markers, CHOP and BiP, but inhibited PKR-like ER kinase and subsequent phosphorylation of eukaryotic initiation factor 2alpha (eif2alpha), both of which are key events in translational suppression. These effects resulted in an accumulation of ubiquitylated proteins leading to protein aggregation and proteotoxicity. Peptide inhibitor or small interfering RNA targeting ER-resident caspase-4 blocked DNA fragmentation, establishing a central role for caspase-4 in bortezomib-induced cell death. The translation inhibitor cycloheximide abrogated bortezomib-induced protein aggregation, caspase-4 processing, and all other characteristics of apoptosis. Because malignant cells have higher protein synthesis rates than normal cells, they may be more prone to protein aggregation and proteotoxicity and possess increased sensitivity to bortezomib-induced apoptosis. Taken together, the results show that bortezomib induces a unique type of ER stress compared with other ER stress agents characterized by an absence of eif2alpha phosphorylation, ubiquitylated protein accumulation, and proteotoxicity.


In vitro culture of embryonic mouse intestinal epithelium: cell differentiation and introduction of reporter genes.
Study of the normal development of the intestinal epithelium has been hampered by a lack of suitable model systems, in particular ones that enable the introduction of exogenous genes. Production of such a system would advance our understanding of normal epithelial development and help to shed light on the pathogenesis of intestinal neoplasia. The criteria for a reliable culture system include the ability to perform real time observations and manipulations in vitro, the preparation of wholemounts for immunostaining and the potential for introducing genes. The new culture system involves growing mouse embryo intestinal explants on fibronectin-coated coverslips in basal Eagle's medium+20% fetal bovine serum. Initially the cultures maintain expression of the intestinal transcription factor Cdx2 together with columnar epithelial (cytokeratin 8) and mesenchymal (smooth muscle actin) markers. Over a few days of culture, differentiation markers appear characteristic of absorptive epithelium (sucrase-isomaltase), goblet cells (Periodic Acid Schiff positive), enteroendocrine cells (chromogranin A) and Paneth cells (lysozyme). Three different approaches were tested to express genes in the developing cultures: transfection, electroporation and adenoviral infection. All could introduce genes into the mesenchyme, but only to a small extent into the epithelium. However the efficiency of adenovirus infection can be greatly improved by a limited enzyme digestion, which makes accessible the lateral faces of cells bearing the Coxsackie and Adenovirus Receptor. This enables reliable delivery of genes into epithelial cells. We describe a new in vitro culture system for the small intestine of the mouse embryo that recapitulates its normal development. The system both provides a model for studying normal development of the intestinal epithelium and also allows for the manipulation of gene expression. The explants can be cultured for up to two weeks, they form the full repertoire of intestinal epithelial cell types (enterocytes, goblet cells, Paneth cells and enteroendocrine cells) and the method for gene introduction into the epithelium is efficient and reliable.


Health and economic burden of obesity in Brazil.
Higher and lower-middle income countries are increasingly affected by obesity. Obesity-related diseases are placing a substantial health and economic burden on Brazil. Our aim is to measure the future consequences of these trends on the associated disease burden and health care costs. A previously developed micro-simulation model is used to project the extent of obesity, obesity-related diseases and associated healthcare costs to 2050. In total, thirteen diseases were considered: coronary heart disease, stroke, hypertension, diabetes, osteoarthritis, and eight cancers. We simulated three hypothetical intervention scenarios: no intervention, 1% and 5% reduction in body mass index (BMI). In 2010, nearly 57% of the Brazilian male population was overweight or obese (BMI ≥25 kg/m(2)), but the model projects rates as high as 95% by 2050. A slightly less pessimistic picture is predicted for females, increasing from 43% in 2010 to 52% in 2050. Coronary heart disease, stroke, hypertension, cancers, osteoarthritis and diabetes prevalence cases are projected to at least double by 2050, reaching nearly 34,000 cases of hypertension by 2050 (per 100,000). 1% and 5% reduction in mean BMI will save over 800 prevalence cases and nearly 3,000 cases of hypertension by 2050 respectively (per 100,000). The health care costs will double from 2010 ($5.8 billion) in 2050 alone ($10.1 billion). Over 40 years costs will reach $330 billion. However, with effective interventions the costs can be reduced to $302 billion by 1% and to $273 billion by 5% reduction in mean BMI across the population. Obesity rates are rapidly increasing creating a high burden of disease and associated costs. However, an effective intervention to decrease obesity by just 1% will substantially reduce obesity burden and will have a significant effect on health care expenditure.


Results of aperture area comparisons for exo-atmospheric total solar irradiance measurements.
Exo-atmospheric solar irradiance measurements made by the solar irradiance community since 1978 have incorporated limiting apertures with diameters measured by a number of metrology laboratories using a variety of techniques. Knowledge of the aperture area is a critical component in the conversion of radiant flux measurements to solar irradiance. A National Aeronautics and Space Administration (NASA) Earth Observing System (EOS) sponsored international comparison of aperture area measurements of limiting apertures provided by solar irradiance researchers was performed, the effort being executed by the National Institute of Standards and Technology (NIST) in coordination with the EOS Project Science Office. Apertures that had institutional heritage with historical solar irradiance measurements were measured using the absolute aperture measurement facility at NIST. The measurement technique employed noncontact video microscopy using high-accuracy translation stages. We have quantified the differences between the participating institutions' aperture area measurements and find no evidence to support the hypothesis that preflight aperture area measurements were the root cause of discrepancies in long-term total solar irradiance satellite measurements. Another result is the assessment of uncertainties assigned to methods used by participants. We find that uncertainties assigned to a participant's values may be underestimated.


Assessment of radiation dose awareness among pediatricians.
There is increasing awareness among pediatric radiologists of the potential risks associated with ionizing radiation in medical imaging. However, it is not known whether there has been a corresponding increase in awareness among pediatricians. To establish the level of awareness among pediatricians of the recent publicity on radiation risks in children, knowledge of the relative doses of radiological investigations, current practice regarding parent/patient discussions, and the sources of educational input. Multiple-choice survey. Of 220 respondents, 105 (48%) were aware of the 2001 American Journal of Roentgenology articles on pediatric CT and radiation, though only 6% were correct in their estimate of the quoted lifetime excess cancer risk associated with radiation doses equivalent to pediatric CT. A sustained or transient increase in parent questioning regarding radiation doses had been noticed by 31%. When estimating the effective doses of various pediatric radiological investigations in chest radiograph (CXR) equivalents, 87% of all responses (and 94% of CT estimates) were underestimates. Only 15% of respondents were familiar with the ALARA principle. Only 14% of pediatricians recalled any relevant formal teaching during their specialty training. The survey response rate was 40%. Awareness of radiation protection issues among pediatricians is generally low, with widespread underestimation of relative doses and risks.


No influence of oxygen levels on pathogenesis and virus shedding in Salmonid alphavirus (SAV)-challenged Atlantic salmon (Salmo salar L.).
For more than three decades, diseases caused by salmonid alphaviruses (SAV) have become a major problem of increasing economic importance in the European fish-farming industry. However, experimental infection trials with SAV result in low or no mortality i.e very different from most field outbreaks of pancreas disease (PD). This probably reflects the difficulties in reproducing complex biotic and abiotic field conditions in the laboratory. In this study we looked at the relationship between SAV-infection in salmon and sub-lethal environmental hypoxia as a result of reduced flow-through in tank systems. The experiment demonstrated that constant reduced oxygen levels (60-65% oxygen saturation: 6.5-7.0 mg/L) did not significantly increase the severity or the progress of pancreas disease (PD). These conclusions are based upon assessments of a semi-quantitative histopathological lesion score system, morbidities/mortalities, and levels of SAV RNA in tissues and water (measured by 1 MDS electropositive virus filters and downstream real-time RT-PCR). Furthermore, we demonstrate that the fish population shed detectable levels of the virus into the surrounding water during viraemia; 4-13 days after i.p. infection, and prior to appearance of severe lesions in heart (21-35 dpi). After this period, viral RNA from SAV could not be detected in water samples although still present in tissues (gills and hearts) at lasting low levels. Lesions could be seen in exocrine pancreas at 7-21 days post infection, but no muscle lesions were seen. In our study, experimentally induced hypoxia failed to explain the discrepancy between the severities reported from field outbreaks of SAV-disease and experimental infections. Reduction of oxygen levels to constant suboptimal levels had no effect on the severity of lesions caused by SAV-infection or the progress of the disease. Furthermore, we present a modified VIRADEL method which can be used to detect virus in water and to supplement experimental infection trials with information related to viral shedding. By using this method, we were able to demonstrate for the first time that shedding of SAV from the fish population into the surrounding water coincides with viraemia.


"Youth as accessories": Stakeholder Perspectives on Youth Participation in Mental Health Policymaking [Part II].
To elicit stakeholder perspectives on the findings from our scoping review on youth participation in mental health policymaking, we conducted a global consultation with young people and adults directly involved in mental health policymaking. Forty-four stakeholders from 16 countries, including 15 young people, 9 policymakers and 20 facilitators of youth participation, took part in individual interviews and/or focus groups. They were asked about how the review findings contrasted with their own experiences in mental health policymaking. The transcribed data were thematically analyzed. All participants viewed lived experience as valuable in identifying policy gaps. Youth pointed out that children and youth with disabilities, diverse sexual orientations, and/or gender identities were often excluded, and spoke about feelings of being an "accessory", illustrating a lack of power-sharing in a tokenized policymaking process. Adult participants' accounts highlighted the challenges inherent in policymaking such as the need for political knowledge and institutional time constraints. A range of cultural, socio-economic, and political barriers to youth participation, that were often context-specific, were identified. The diverse perspectives of stakeholders extended the review results. Based on our findings, we recommend that adults and institutions: (1) recognize lived experience as expertise in shaping mental health policies; (2) include diverse groups; (3) reduce tokenistic relationships through the creation of safer spaces, adult feedback, co-production, and social accountability; and (4) adopt an intersectional approach to address cultural, socio-economic, and political barriers to participation. Methodologically, our work demonstrates why stakeholder consultations are an essential component of scoping reviews.


Dual endothelin-converting enzyme/neutral endopeptidase blockade in rats with D-galactosamine-induced liver failure.
Secondary activation of the endothelin system is thought to be involved in toxic liver injury. This study tested the hypothesis that dual endothelin-converting enzyme / neutral endopeptidase blockade might be able to attenuate acute toxic liver injury. - Male Sprague-Dawley rats were implanted with subcutaneous minipumps to deliver the novel compound SLV338 (10 mg/kg*d) or vehicle. Four days later they received two intraperitoneal injections of D-galactosamine (1.3 g/kg each) or vehicle at an interval of 12 hours. The animals were sacrificed 48 hours after the first injection. - Injection of D-galactosamine resulted in very severe liver injury, reflected by strongly elevated plasma liver enzymes, hepatic necrosis and inflammation, and a mortality rate of 42.9 %. SLV338 treatment did not show any significant effect on the extent of acute liver injury as judged from plasma parameters, hepatic histology and mortality. Plasma measurements of SLV338 confirmed adequate drug delivery. Plasma concentrations of big endothelin-1 and endothelin-1 were significantly elevated in animals with liver injury (5-fold and 62-fold, respectively). Plasma endothelin-1 was significantly correlated with several markers of liver injury. SLV338 completely prevented the rise of plasma big endothelin-1 (p<0.05) and markedly attenuated the rise of endothelin-1 (p = 0.055). - In conclusion, dual endothelin-converting enzyme / neutral endopeptidase blockade by SLV338 did not significantly attenuate D-galactosamine-induced acute liver injury, although it largely prevented the activation of the endothelin system. An evaluation of SLV338 in a less severe model of liver injury would be of interest, since very severe intoxication might not be relevantly amenable to pharmacological interventions.


Jaw fractures in Nigerian children: an analysis of 102 cases.
To present the prevalence and pattern of jaw fractures in children aged 15 years and below attending two hospitals serving as referral centres for facial injuries in North Eastern Nigeria. Retrospective Cross Sectional Study. Department of Oral and Maxillofacial Surgery, University of Maiduguri Teaching Hospital, Borno State, Nigeria. One hundred and two patients with jaw fractures aged 15 years and below. Aetiological factors and pattern of jaw fractufes in children aged 15 years and below. We retrospectively reviewed 102 patients with jaw fractures aged 15 years and below seen over a five year period at two referral centres in Maiduguri, Northeastern Nigeria. This constituted 9.5% of the total 1,074 cases of maxillofacial injuries managed during the period. The male to female ratio was 7.5:1 and there was a male responderance in all age groups. The main causes of fractures were road traffic accident (n = 55, 53.7%) followed by falls (n = 26, 25%). Other etiological factors such as fights, sports and gunshots accounted for the remaining cases. The mandible was the commonest site with the body commonly fractured (n = 74, 72.5%). No Le Fort III fractures were identified in this study. Despite some limitations to our data, such as exclusion of patients who attended private clinics and lack of modern diagnostic methods, this report shows that there is obvious need for concern about the high prevalence of maxillofacial injuries in children caused by road traffic accidents in North Eastern Nigeria.


Isolation and rapid sharing of the 2019 novel coronavirus (SARS-CoV-2) from the first patient diagnosed with COVID-19 in Australia.
To describe the first isolation and sequencing of SARS-CoV-2 in Australia and rapid sharing of the isolate. SARS-CoV-2 was isolated from a 58-year-old man from Wuhan, China who arrived in Melbourne on 19 January 2020 and was admitted to the Monash Medical Centre, Melbourne from the emergency department on 24 January 2020 with fever, cough, and progressive dyspnoea. Clinical course and laboratory features of the first reported case of COVID-19 (the illness caused by SARS-CoV-2) in Australia; isolation, whole genome sequencing, imaging, and rapid sharing of virus from the patient. A nasopharyngeal swab and sputum collected when the patient presented to hospital were each positive for SARS-CoV-2 (reverse transcription polymerase chain reaction). Inoculation of Vero/hSLAM cells with material from the nasopharyngeal swab led to the isolation of SARS-CoV-2 virus in culture. Electron microscopy of the supernatant confirmed the presence of virus particles with morphology characteristic of viruses of the family Coronaviridae. Whole genome sequencing of the viral isolate and phylogenetic analysis indicated the isolate exhibited greater than 99.99% sequence identity with other publicly available SARS-CoV-2 genomes. Within 24 hours of isolation, the first Australian SARS-CoV-2 isolate was shared with local and overseas reference laboratories and major North American and European culture collections. The ability to rapidly identify, propagate, and internationally share our SARS-CoV-2 isolate is an important step in collaborative scientific efforts to deal effectively with this international public health emergency by developing better diagnostic procedures, vaccine candidates, and antiviral agents.


Nuclear magnetic resonance study of regional metabolism after forebrain ischemia in rats.
Proton nuclear magnetic resonance (NMR) spectroscopy of perchloric acid tissue extracts has been used to follow serial postischemic changes in the levels of metabolites in the hippocampus, cerebellum, frontal lobes, and parietal/occipital lobes in a rat model of short-duration (10 minutes) forebrain ischemia. Shortly (10 minutes, 1 hour) after the ischemic insult, the levels of the amino acids alanine and gamma-aminobutyric acid are elevated and that of glutamate is depressed in all regions except the cerebellum. The levels of these species return to control values by 24 hours postischemia. No changes are observed in the levels of aspartate or N-acetylaspartate. Greatly elevated levels of acetate 10 minutes postischemia, particularly in the hippocampus, may be due in part to metabolic degradation of fatty acids released due to membrane breakdown. Elevated levels of lactate persist for up to 7 days postischemia, suggesting that normal mitochondrial functioning is not fully restored following the ischemic insult.


Pediatric Condition Falsification Misdiagnosed by Misjudged Weight Growth from the Curve of Measured Weights.
BACKGROUND Pediatric condition falsification (PCF) is a rare form of child abuse in which a caregiver fabricates or induces illness in the child. The diagnosis is difficult and controversial and can easily include false positives. CASE REPORT A boy, 3.18 kg birthweight (P25 curve), lost weight between age 56 to120 days. Cow milk allergy was suspected, feeding was changed to elementary formula, and he started catch-up weight growth while remaining significantly underweight. His pediatrician continuously interpreted his low weight as insufficient growth, despite prescribing 3 times the normal caloric intake, concluded that the mother purposely malnourished her son, diagnosed PCF, and the boy was separated from his family (days 502-755 of age). PCF was confirmed by 2 other pediatricians and 3 child protection physicians and was supported by 4 child protection agencies and 6 judges. However, proper analysis of the weight growth (kg/year) from the weight curve showed a normal weight gain. Beyond 120 days of age, weight gain at home was significantly above normal (during 347-489 days: 6.2 versus 3 kg/year of the P50). He reached P25 again at around 516 days. CONCLUSIONS The question "How could so many physicians misjudge weight gain?" has scientific and sociologic aspects. Scientifically, low weight was wrongly interpreted as insufficient weight growth, requiring that physicians learn how to assess weight gain from weight curves. Sociologically, physicians seem to follow a diagnosis made by a colleague without proper evaluation. Arguments provided by the parents against this diagnosis seemed to be neglected. Confirmation bias occurs when any information against PCF is disregarded.


Analysis of Baseline Computerized Neurocognitive Testing Results among 5-11-Year-Old Male and Female Children Playing Sports in Recreational Leagues in Florida.
There is a paucity of data related to sports injuries, concussions, and computerized neurocognitive testing (CNT) among very young athletes playing sports in recreational settings. The purpose of this study was to report baseline CNT results among male and female children, ages 5-11, playing sports in Hillsborough County, Florida using ImPACT Pediatric, which is specifically designed for this population. Data were collected from 2016 to 2017. The results show that 657 baseline tests were conducted and t-tests and linear regression were used to assess mean significant differences in composite scores with sex and age. Results showed that females scored better on visual memory and in general as age increased, baseline scores improved. The results can be used to build further studies on the use of CNT in recreational settings and their role in concussion treatment, management, and interventions.


Sexual partner preference, hypothalamic morphology and aromatase in rams.
The male-oriented ram is a unique and valuable animal model for the study of hormonal, developmental and genetic contributions to sexual partner preference. Unlike most other mammalian models that are in use currently, variations in sexual attraction occur spontaneously in domestic ram populations. It is estimated that as many as 8-10% of rams exhibit a sexual partner preference for other males, classifying them as male-oriented rams. Studies have failed to identify any compelling social factors that can predict or explain the variations in sexual partner preferences of rams. Nor is there consensus on the endocrine and sensory responsiveness of male-oriented rams to other rams. However, a number of studies have reported differences in brain structure and function between male-oriented and female-oriented rams, suggesting that sexual partner preferences are neurologically hard-wired. Recently, we identified a sexually dimorphic nucleus (oSDN) in the sheep preoptic area-anterior hypothalamus. The oSDN is larger in female-oriented rams than in male-oriented rams and similar in size in male-oriented rams and ewes. In addition, mRNA levels for aromatase in the oSDN were higher in males than in females and were higher in female-oriented rams than in male-oriented rams. These results suggest a relationship between steroid hormones, specifically estrogens and oSDN morphology. In this review, we provide an overview of sexual behavior in rams and discuss the multiple factors that may contribute to the development and adult expression of same-sex partner preferences in rams.


Influenza Virus with Increased pH of Hemagglutinin Activation Has Improved Replication in Cell Culture but at the Cost of Infectivity in Human Airway Epithelium.
Pandemic H1N1 (pH1N1) influenza virus emerged from swine in 2009 with an adequate capability to infect and transmit between people. In subsequent years, it has circulated as a seasonal virus and evolved further human-adapting mutations. Mutations in the hemagglutinin (HA) stalk that increase pH stability have been associated with human adaptation and airborne transmission of pH1N1 virus. Yet, our understanding of how pH stability impacts virus-host interactions is incomplete. Here, using recombinant viruses with point mutations that alter the pH stability of pH1N1 HA, we found distinct effects on virus phenotypes in different experimental models. Increased pH sensitivity enabled viruses to uncoat in endosomes more efficiently, manifesting as increased replication rate in typical continuous cell cultures under single-cycle conditions. A more acid-labile HA also conferred a small reduction in sensitivity to antiviral therapeutics that act at the pH-sensitive HA fusion step. Conversely, in primary human airway epithelium cultured at the air-liquid interface, increased pH sensitivity attenuated multicycle viral replication by compromising virus survival in the extracellular microenvironment. In a mouse model of influenza pathogenicity, there was an optimum HA activation pH, and viruses with either more- or less-pH-stable HA were less virulent. Opposing pressures inside and outside the host cell that determine pH stability may influence zoonotic potential. The distinct effects that changes in pH stability exert on viral phenotypes underscore the importance of using the most appropriate systems for assessing virus titer and fitness, which has implications for vaccine manufacture, antiviral drug development, and pandemic risk assessment.IMPORTANCE The pH stability of the hemagglutinin surface protein varies between different influenza strains and subtypes and can affect the virus' ability to replicate and transmit. Here, we demonstrate a delicate balance that the virus strikes within and without the target cell. We show that a pH-stable hemagglutinin enables a human influenza virus to replicate more effectively in human airway cells and mouse lungs by facilitating virus survival in the extracellular environment of the upper respiratory tract. Conversely, after entering target cells, being more pH stable confers a relative disadvantage, resulting in less efficient delivery of the viral genome to the host cell nucleus. Since the balance we describe will be affected differently in different host environments, it may restrict a virus' ability to cross species. In addition, our findings imply that different influenza viruses may show variation in how well they are controlled by antiviral strategies targeting pH-dependent steps in the virus replication cycle.


Deep direct likelihood knockoffs.
Predictive modeling often uses black box machine learning methods, such as deep neural networks, to achieve state-of-the-art performance. In scientific domains, the scientist often wishes to discover which features are actually important for making the predictions. These discoveries may lead to costly follow-up experiments and as such it is important that the error rate on discoveries is not too high. Model-X knockoffs [2] enable important features to be discovered with control of the false discovery rate (fdr). However, knockoffs require rich generative models capable of accurately modeling the knockoff features while ensuring they obey the so-called "swap" property. We develop Deep Direct Likelihood Knockoffs (ddlk), which directly minimizes the KL divergence implied by the knockoff swap property. ddlk consists of two stages: it first maximizes the explicit likelihood of the features, then minimizes the KL divergence between the joint distribution of features and knockoffs and any swap between them. To ensure that the generated knockoffs are valid under any possible swap, ddlk uses the Gumbel-Softmax trick to optimize the knockoff generator under the worst-case swap. We find ddlk has higher power than baselines while controlling the false discovery rate on a variety of synthetic and real benchmarks including a task involving a large dataset from one of the epicenters of COVID-19.


Measuring Treatment Integrity: Use of and Experience with Measurements in Child and Youth Care Organizations.
Performance feedback and supervision are essential to the adoption of evidence-based interventions with high treatment integrity in child and youth care organizations. Little is known about the use of treatment integrity measurements in these organizations. For this study, 12 interventions for children and young people in the Netherlands with externalizing behavioral problems were selected. For each intervention, an expert, two supervisors, and two therapists were approached for an interview. In total, 54 semi-structured interviews were conducted. The results show that almost all interventions used treatment integrity instruments (N = 11, 91.7%). Only two used measurements for both QA procedures (certification and recertification) and supervision purposes. Therapists regard treatment integrity measurements as valuable when they are used for multiple purposes and feedback is provided. The results of this study suggest the feasibility of the use of measurements for multiple purposes. Collaborative action is required to develop instruments that effectively contribute to continuous improvement.


A simplified method to estimate Diphyllobothrium spp. infection in salmonids.
Some fish parasites constitute severe management problems as they may cause mortality of their fish host or are important zoonoses of humans. Parasite assessments are therefore critical to keep track of infections. If conventional sampling techniques can be simplified, parasite assessments might be easier to obtain, less time-consuming and more extensive. In this study, we compare the assessed number of Diphyllobothrium spp. cysts (CYST) with the counted number of Diphyllobothrium spp. plerocercoid larvae recovered using a conventional digestive technique (LARV). The aim was to determine the potential of using CYST as a simplified methodology for assessing Diphyllobothrium spp. infection in salmonids. In total, 365 brown trout and 424 Arctic charr were sampled from nine lakes in subarctic Norway. Strong correlation, significant linear relationship and large amount of explained variation were found between log10 CYST and log10 LARV in both fish species. The method had a slight, but not significant tendency to work better in charr compared to trout. In addition, absolute difference between CYST and LARV increased at parasite intensities >100 indicating that the method has reduced functionality when estimating parasite intensity in heavily infected salmonid populations. However, overall, using this simplified and less time-consuming methodology, a good indication of Diphyllobothrium spp. intensity, abundance and prevalence was obtained. We suggest that this method provides a sound proxy of the Diphyllobothrium spp. burden and have the potential to be used in parasite assessment during fish monitoring and fisheries management surveys, particularly if the time and resources for detailed parasite studies are not available.


A Rodent Model of Exposure Therapy: The Use of Fear Extinction as a Therapeutic Intervention for PTSD.
The symptoms of post-traumatic stress disorder (PTSD) include cognitive impairment related to medial prefrontal cortical dysfunction. Indeed, a deficit of cognitive flexibility, i.e., an inability to modify previously learned thoughts and behaviors based on changes in the environment, may underlie many of the other symptoms of PTSD, such as changes in mood, hyper-arousal, intrusive thoughts, exaggerated and over-generalized fear, and avoidance behavior. Cognitive-behavioral therapies target the cognitive dysfunction observed in PTSD patients, training them to recalibrate stress-related perceptions, interpretations and responses. Preclinically, the extinction of conditioned fear bears resemblance to one form of cognitive therapy, exposure therapy, whereby an individual learns, through repeated exposure to a fear-provoking stimulus in a safe environment, that the stimulus no longer signals imminent threat, and their fear response is suppressed. In this review article, we highlight recent findings from our lab using fear extinction as a preclinical model of exposure therapy in rodents exposed to chronic unpredictable stress (CUS). We specifically focus on the therapeutic effects of extinction on stress-compromised set-shifting as a measure of cognitive flexibility, and active vs. passive coping behavior as a measure of avoidance. Finally, we discuss mechanisms involving activity and plasticity in the medial prefrontal cortex (mPFC) necessary for the therapeutic effects of extinction on cognitive flexibility and active coping.


Nitrate variability in groundwater of North Carolina using monitoring and private well data models.
Nitrate (NO3-) is a widespread contaminant of groundwater and surface water across the United States that has deleterious effects to human and ecological health. This study develops a model for predicting point-level groundwater NO3- at a state scale for monitoring wells and private wells of North Carolina. A land use regression (LUR) model selection procedure is developed for determining nonlinear model explanatory variables when they are known to be correlated. Bayesian Maximum Entropy (BME) is used to integrate the LUR model to create a LUR-BME model of spatial/temporal varying groundwater NO3- concentrations. LUR-BME results in a leave-one-out cross-validation r2 of 0.74 and 0.33 for monitoring and private wells, effectively predicting within spatial covariance ranges. Results show significant differences in the spatial distribution of groundwater NO3- contamination in monitoring versus private wells; high NO3- concentrations in the southeastern plains of North Carolina; and wastewater treatment residuals and swine confined animal feeding operations as local sources of NO3- in monitoring wells. Results are of interest to agencies that regulate drinking water sources or monitor health outcomes from ingestion of drinking water. Lastly, LUR-BME model estimates can be integrated into surface water models for more accurate management of nonpoint sources of nitrogen.


Production and Properties of Xylan-Degrading Enzymes from Cellulomonas uda.
Xylan degradation and production of beta-xylanase and beta-xylosidase activities were studied in cultures of Cellulomonas uda grown on purified xylan from birchwood. beta-Xylanase activity was found to be associated with the cells, although in various degrees. The formation of beta-xylanase activity was induced by xylotriose and repressed by xylose. beta-Xylosidase activity was cell bound. Both constitutive and inducible beta-xylosidase activities were suggested. beta-Xylanase and beta-xylosidase activities were inhibited competitively by xylose. beta-Xylanase activity had a pronounced optimum pH of 5.8, whereas the optimum pH of beta-xylosidase activity ranged from 5.4 to 6.1. The major products of xylan degradation by a crude preparation of beta-xylanase activity, in decreasing order of amount, were xylobiose, xylotriose, xylose, and small amounts of xylotetraose. This pattern suggests that beta-xylanase activity secreted by C. uda is of the endosplitting type. Supernatants of cultures grown on cellulose showed not only beta-glucanase but also beta-xylanase activity. The latter could be attributed to an endo-1,4-beta-glucanase activity which had a low beta-xylanase activity.


Migration of toxoplasma gondii-infected dendritic cells across human retinal vascular endothelium.
Toxoplasma gondii, the parasite responsible for ocular toxoplasmosis, accesses the retina from the bloodstream. We investigated the dendritic cell as a potential taxi for T. gondii tachyzoites moving across the human retinal endothelium, and examined the participation of adhesion molecules and chemokines in this process. CD14-positive monocytes were isolated from human peripheral blood by antibody-mediated cell enrichment, and cultured in granulocyte-macrophage colony-stimulating factor and interleukin-4 to generate dendritic cells. Transmigration assays were performed over 18 hours in transwells seeded with human retinal endothelial cells and using dendritic cells exposed to laboratory or natural strains of T. gondii tachyzoites. Parasites were tagged with yellow fluorescent protein to verify infection. In some experiments, endothelial monolayers were preincubated with antibody directed against adhesion molecules, or chemokine was added to lower chambers of transwells. Human monocyte-derived dendritic cell preparations infected with laboratory or natural strain T. gondii tachyzoites transmigrated in larger numbers across simulated human retinal endothelium than uninfected dendritic cells (P ≤ 0.0004 in 5 of 6 experiments). Antibody blockade of intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, and activated leukocyte cell adhesion molecule (ALCAM) inhibited transmigration (P ≤ 0.007), and CCL21 or CXCL10 increased transmigration (P ≤ 0.031). Transmigration of human dendritic cells across retinal endothelium is increased following infection with T. gondii. Movement may be impacted by locally produced chemokines and is mediated in part by ICAM-1, VCAM-1, and ALCAM. These findings have implications for development of novel therapeutics aimed at preventing retinal infection by T. gondii.


Calcium channel receptor sites for (+)-[3H]PN 200-110 in coronary artery.
The receptor sites for 1,4-dihydropyridine (DHP) Ca++ channel antagonists in porcine coronary artery were identified and characterized by a binding assay using (+)-[3H]PN 200-110 as a radioligand. Specific (+)-[3H]PN 200-110 binding in porcine coronary artery was saturable, reversible and of high affinity (Kd = 0.24 nM) and it showed a pharmacological specificity as well as stereoselectivity which characterized the receptor sites for DHP Ca++ channel antagonists. DHP antagonists competed for the (+)-[3H]PN 200-110 binding in order: PN 200-110 greater than mepirodipine greater than nisoldipine greater than nicardipine greater than nitrendipine greater than nimodipine greater than nifedipine greater than (-)-PN 200-110. (+)-PN 200-110 was approximately 140 times as potent as the (-)-isomer. The potencies (PKi) of these eight DHP Ca++ channel antagonists in competing for (+)-[3H]PN 200-110 binding sites in porcine coronary artery correlated well with their pharmacological potencies. Specific (+)-[3H]PN 200-110 binding in the coronary artery was enhanced by d-cis-diltiazem and was inhibited incompletely by verapamil and D-600. In EDTA-pretreated coronary artery, the maximal number of binding sites for specific (+)-[3H]PN 200-110 binding was reduced (80%) markedly, and it was restored to the untreated level by the addition of Ca++ and Mg++.(ABSTRACT TRUNCATED AT 250 WORDS)


Intraosseous infantile myofibroma of the mandible.
The myofibroma, especially the intraosseous variety, is an uncommon benign tumor that occurs primarily in children younger than 3 years of age. Since 1966, less than 40 cases of solitary myofibromas of the mandible have been reported in the literature. Myofibroblasts and spindle cells are predominantly found in this benign lesion. These cells are also commonly found in many lesions due to which great difficulty can be encountered in the diagnosis. A rare case of the intraosseous variety of an infantile myofibroma of the mandible diagnosed in a 10-month-old child has been reported. The tumor was completely excised via an intraoral approach and no recurrence has been noted 15 months postoperatively. A brief review of the differential diagnosis of this lesion in terms of its clinicopathologic, histologic, and immunohistochemical features is also discussed.


The Role of Surgery for Asymptomatic Primary Tumors in Unresectable Stage IV Colorectal Cancer.
There are still debates regarding the appropriate primary treatment policy for asymptomatic primary colorectal lesions in cases of unresectable metastatic colorectal cancer. Even though there are patients with asymptomatic primary tumors when starting chemotherapy, those patients may still undergo surgery due to complications related to primary tumors in the middle of chemotherapy; therefore, controversy exists regarding surgical resection of primary colorectal lesions in cases where symptoms are absent when making a diagnosis. Thus, based on the published literature, we discuss opinions that prefer first-line surgery for primary tumors as well as opinions favoring first-line chemotherapy for treating unresectable synchronous metastatic colorectal cancer. Although the upfront chemotherapy including targeted agents is suggested as an effective treatment in recent years, the first line surgery has been a preferred treatment for decades. The first line surgery is beneficial to prolong the survival duration given the retrospective analysis of randomized trial data. So far, no prospective comparison study has only focused on the first-line treatment modality; thus, future clinical studies focusing on the survival duration and the quality of life should be performed as soon as possible. Furthermore, at this point, multidisciplinary team approaches would be helpful in finding the appropriate therapy. Regardless of symptoms, the performance status and the tumor burden should be taken into consideration as well. In case of surgical resection, minimally invasive surgery, such as laparoscopic surgery, is recommended.


Numerical validation of a new method to assess aortic pulse wave velocity from a single recording of a brachial artery waveform with an occluding cuff.
Recently a new method has been proposed as a tool to measure arterial pulse wave velocity (PWV), a measure of the stiffness of the large arteries and an emerging parameter used as indicator of clinical cardiovascular risk. The method is based on measurement of brachial blood pressure during supra-systolic pressure inflation of a simple brachial cuff [the device is known as the Arteriograph (Tensiomed, Budapest, Hungary)]. This occlusion yields pronounced first and secondary peaks in the pressure waveform, the latter ascribed to a reflection from the aortic bifurcation, and PWV is calculated as the ratio of twice the jugulum-symphysis distance and the time difference between the two peaks. To test the validity of this working principle, we used a numerical model of the arterial tree to simulate pressures and flows in the normal configuration, and in a configuration with an occluded brachial artery. A pronounced secondary peak was indeed found in the brachial pressure signal of the occluded model, but its timing was only related to brachial stiffness and not to aortic stiffness. We also compared PWV's calculated with three different methods: PWVATG (approximately Arteriograph principle), PWVcar-fem (approximately carotid-femoral PWV, the current clinical gold standard method), and PWVtheor (approximately Bramwell-Hill equation). Both PWVATG (R2=0.94) and PWVcar-fem (R2=0.95) correlated well with PWVtheor, but their numerical values were lower (by 2.17+/-0.42 and 1.08+/-0.70 m/s for PWVATG and PWVcar-fem, respectively). In conclusion, our simulations question the working principle of the Arteriograph. Our data indicate that the method picks up wave reflection phenomena confined to the brachial artery, and derived values of PWV rather reflect the stiffness of the brachial arteries.


Differentiated parietal connectivity of frontal regions for "what" and "where" memory.
In a previous meta-analysis across almost 200 neuroimaging experiments, working memory for object location showed significantly stronger convergence on the posterior superior frontal gyrus, whereas working memory for identity showed stronger convergence on the posterior inferior frontal gyrus (dorsal to, but overlapping with Brodmann's area BA 44). As similar locations have been discussed as part of a dorsal frontal-superior parietal reach system and an inferior frontal grasp system, the aim of the present study was to test whether the regions of working-memory related "what" and "where" processing show a similar distinction in parietal connectivity. The regions that were found in the previous meta-analysis were used as seeds for functional connectivity analyses using task-based meta-analytic connectivity modelling and task-independent resting state correlations. While the ventral seed showed significantly stronger connectivity with the bilateral intraparietal sulcus (IPS), the dorsal seed showed stronger connectivity with the bilateral posterior inferior parietal and the medial superior parietal lobule. The observed connections of regions involved in memory for object location and identity thus clearly demonstrate a distinction into separate pathways that resemble the parietal connectivity patterns of the dorsal and ventral premotor cortex in non-human primates and humans. It may hence be speculated that memory for a particular location and reaching towards it as well as object memory and finger positioning for manipulation may rely on shared neural systems. Moreover, the ensuing regions, in turn, featured differential connectivity with the bilateral ventral and dorsal extrastriate cortex, suggesting largely segregated bilateral connectivity pathways from the dorsal visual cortex via the superior and inferior parietal lobules to the dorsal posterior frontal cortex and from the ventral visual cortex via the IPS to the ventral posterior frontal cortex that may underlie action and cognition.


Genetic diversity of human head lice and molecular detection of associated bacterial pathogens in Democratic Republic of Congo.
Head louse, Pediculus humanus capitis, is an obligatory blood-sucking ectoparasite, distributed worldwide. Phylogenetically, it occurs in five divergent mitochondrial clades (A-E); each exhibiting a particular geographical distribution. Recent studies suggest that, as in the case of body louse, head louse could be a disease vector. We aimed to study the genetic diversity of head lice collected in the Democratic Republic of the Congo (DR Congo) and to screen for louse-borne pathogens in these lice. A total of 181 head lice were collected from 27 individuals at the Monkole Hospital Center located in Kinshasa. All head lice were genotyped and screened for the presence of louse-borne bacteria using molecular methods. We searched for Bartonella quintana, Borrelia recurrentis, Rickettsia prowazekii, Anaplasma spp., Yersinia pestis, Coxiella burnetii and Acinetobacter spp. Among these head lice, 67.4% (122/181) belonged to clade A and 24.3% (44/181) belonged to clade D. Additionally, for the first time in this area, we found clade E in 8.3% (15/181) of tested lice, from two infested individuals. Dual infestation with clades A and D was observed for 44.4% individuals. Thirty-three of the 181 head lice were infected only by different bacterial species of the genus Acinetobacter. Overall, 16 out of 27 individuals were infested (59.3%). Six Acinetobacter species were detected including Acinetobacter baumannii (8.3%), Acinetobacter johnsonii (1.7%), Acinetobacter soli (1.7%), Acinetobacter pittii (1.7%), Acinetobacter guillouiae (1.1%), as well as a new potential species named "Candidatus Acinetobacter pediculi". To our knowledge, this study reports for the first time, the presence of clade E head lice in DR Congo. This study is also the first to report the presence of Acinetobacter species DNAs in human head lice in DR Congo.


A brain-computer interface based attention training program for treating attention deficit hyperactivity disorder.
Attention deficit hyperactivity disorder (ADHD) symptoms can be difficult to treat. We previously reported that a 20-session brain-computer interface (BCI) attention training programme improved ADHD symptoms. Here, we investigated a new more intensive BCI-based attention training game system on 20 unmedicated ADHD children (16 males, 4 females) with significant inattentive symptoms (combined and inattentive ADHD subtypes). This new system monitored attention through a head band with dry EEG sensors, which was used to drive a feed forward game. The system was calibrated for each user by measuring the EEG parameters during a Stroop task. Treatment consisted of an 8-week training comprising 24 sessions followed by 3 once-monthly booster training sessions. Following intervention, both parent-rated inattentive and hyperactive-impulsive symptoms on the ADHD Rating Scale showed significant improvement. At week 8, the mean improvement was -4.6 (5.9) and -4.7 (5.6) respectively for inattentive symptoms and hyperactive-impulsive symptoms (both p<0.01). Cohen's d effect size for inattentive symptoms was large at 0.78 at week 8 and 0.84 at week 24 (post-boosters). Further analysis showed that the change in the EEG based BCI ADHD severity measure correlated with the change ADHD Rating Scale scores. The BCI-based attention training game system is a potential new treatment for ADHD. ClinicalTrials.gov NCT01344044.


Lower extremity computed tomography angiography can help predict technical success of endovascular revascularization in the superficial femoral and popliteal artery.
Preprocedural computed tomography angiography (CTA) assists in evaluating vascular morphology and disease distribution and in treatment planning for patients with lower extremity peripheral artery disease (PAD). The aim of the study was to determine the predictive value of radiographic findings on CTA and technical success of endovascular revascularization of occlusions in the superficial femoral artery-popliteal (SFA-pop) region. Medical records and available imaging studies were reviewed for patients undergoing endovascular intervention for PAD between January 2013 and December 2015 at a single academic institution. Radiologists reviewed preoperative CTA scans of patients with occlusions in the SFA-pop region. Radiographic criteria previously used to evaluate chronic occlusions in the coronary arteries were used. Technical success, defined as restoration of inline flow through the SFA-pop region with <30% stenosis at the end of the procedure, and intraoperative details were evaluated. From 2013 to 2015, there were 407 patients who underwent 540 endovascular procedures for PAD. Preprocedural CTA scans were performed in 217 patients (53.3%), and 84 occlusions in the SFA-pop region were diagnosed. Ten occlusions were excluded as no endovascular attempt to cross the lesion was made because of extensive disease or concomitant iliac intervention. Of the remaining 74 occlusions in the SFA-pop region, 59 were successfully treated (80%) and 15 were unsuccessfully crossed (20%). The indications for revascularization were claudication in 57% of patients and critical limb ischemia in the remaining patients. TransAtlantic Inter-Society Consensus A, B, and C occlusions were treated with 87% success, whereas D occlusions were treated with 68% success (P = .047). There were nine occlusions with 100% vessel calcification that was associated with technical failure (P = .014). Longer lengths of occlusion were also associated with technical failure (P = .042). Multiple occlusions (P = .55), negative remodeling (P = .69), vessel runoff (P = .56), and percentage of vessel calcification (P = .059) were not associated with failure. On multivariable analysis, 100% calcification remained the only significant predictor of technical failure (odds ratio, 9.0; 95% confidence interval, 1.8-45.8; P = .008). Analysis of preoperative CTA shows 100% calcification as the best predictor of technical failure of endovascular revascularization of occlusions in the SFA-pop region. Further studies are needed to determine the cost-effectiveness of obtaining preoperative CTA for lower extremity PAD.


Studies on the maturation of the head of bacteriophage T4.
The presentation focuses on the structural rearrangements of the subunits and the processing of the various protein constituents which accompany the maturation events of the head of bacteriophage T4. The major features of the maturation steps of the head are the following: (a) the viral DNA is pulled into an empty head in a series of events; (b) cleavage of two core proteins, P22 (mol. mass = 31000), to small fragments and the internal protein IPIII (mol. mass = 23000) to IPIII (mol. mass = 21000) appears to be intimately linked to the DNA packaging event, whereas the cleavage of the major head protein of the viral coat, P23 (mol. mass = 55000), to P23 (mol. mass = 45000) precedes the DNA packaging event. Recently, we have obtained information about the mechanism by which the viral DNA is pulled into a preformed empty head. Our evidence suggests that the DNA becomes attached to the inside of the empty head and is subsequently collapsed in the interior by the so-called internal peptides. These are highly acidic and derived from a large precursor protein by cleavage.


'Candidatus Halomonas phosphatis', a novel polyphosphate-accumulating organism in full-scale enhanced biological phosphorus removal plants.
Microautoradiography combined with fluorescence in situ hybridization (MAR-FISH) was used to screen for potential polyphosphate-accumulating organisms (PAOs) in full-scale enhanced biological phosphorus removal (EBPR) plants. Clone library analyses and application of MAR-FISH using newly designed probes revealed that small rods related to uncultured Halomonas within the gammaproteobacterial family Halomonadaceae were actively involved in uptake of orthophosphate. Although deeply branched in the Gammaproteobacteria, they were not targeted by the gammaproteobacterial probe (GAM42a). A part of them were also not targeted with the general bacterial probes (EUBmix). They could take up short-chain fatty acids (e.g. acetate and propionate) and ethanol under both anaerobic and aerobic conditions. Polyhydroxyalkanoate storage was observed under anaerobic conditions. There was no indication of a denitrifying capability. A survey of the occurrence of these Halomonas-PAOs in 23 full-scale EBPR plants revealed that they made up 0.5-5.7% of all bacteria in the plants, and were often in higher abundance than the well-described PAOs 'Candidatus Accumulibacter phosphatis'. This indicates a potentially important role for these uncultured Halomonas bacteria in the EBPR process in full-scale plants and we propose to name them 'Candidatus Halomonas phosphatis'.


Phase III, multicentre, randomised, double-blind, placebo-controlled, 104-week study of subcutaneous belimumab administered in combination with rituximab in adults with systemic lupus erythematosus (SLE): BLISS-BELIEVE study protocol.
Belimumab, an anti-B-lymphocyte-stimulator antibody, is approved for the treatment of active, autoantibody-positive systemic lupus erythematosus (SLE). Rituximab, a B cell-depleting anti-CD20 antibody, remains in the SLE treatment armamentarium despite failed trials in lupus nephritis and extrarenal lupus. These biologics, which operate through complementary mechanisms, might result in an enhanced depletion of circulating and tissue-resident autoreactive B lymphocytes when administered together. Thus, belimumab and rituximab combination may be a highly effective treatment of SLE. This study aims to evaluate and compare the efficacy, safety and tolerability of subcutaneous (SC) belimumab and a single cycle of rituximab in patients with SLE with belimumab alone. BLISS-BELIEVE is a three-arm, randomised, double-blind, placebo-controlled, 104-week superiority study. Two hundred adults with SLE will be randomised 1:2:1 to arm A, belimumab SC 200 mg/week for 52 weeks plus placebo at weeks 4 and 6; arm B, belimumab SC 200 mg/week for 52 weeks plus rituximab 1000 mg at weeks 4 and 6; arm C, belimumab SC 200 mg/week plus standard of care for 104 weeks. The 52-week treatment period (arms A and B) is followed by a 52-week observational phase. The primary efficacy endpoint is the proportion of patients with disease control (SLE Disease Activity Index (SLEDAI)-2K≤2, without immunosuppressants and with a prednisone-equivalent dose of ≤5 mg/day) at week 52. Major secondary efficacy endpoints are the proportion of patients in clinical remission (defined as SLEDAI-2K=0, without immunosuppressants and corticosteroids) at week 64, and the proportion of patients with disease control at week 104. Safety endpoints include the incidence of adverse events (AEs), serious AEs and AEs of special interest. Within 6 months of the study's primary manuscript publication, anonymised individual participant data and study documents can be requested for further research from www.clinicalstudydatarequest.com. NCT03312907; Pre-results.


The iron-proximal histidine linkage and protein control of oxygen binding in hemoglobin. A transient Raman study.
Time-resolved Raman studies have shown that communication between the heme oxygen binding sites and the surrounding globin occurs through the iron-proximal histidine linkage. By comparing the frequency of the Fe-His stretching mode in equilibrium deoxy- and photoinduced transient deoxyhemoglobins, we have found that ligand binding induces protein structural changes that strengthen the Fe-His linkage. The extent of this increase is observed to depend upon the quaternary state. This dependence is reflected in the restricted set of values observed in transient and equilibrium studies for the frequency of the Fe-His stretching mode in both R and T state deoxyhemoglobins. For T state hemoglobins the frequency increases from 215 cm-1 to approximately 222 cm-1 in going from the stable to the nanosecond transient deoxy species. The corresponding change for R state human hemoglobins is from approximately 222 cm-1 to approximately 230 cm-1. Studies on transients derived from Fe-Co hybrid hemoglobins reveal no subunit heterogeneity associated with the R state transient (230 cm-1) at high pH. We also observe that perturbations of the protein known to destabilize the structure of ligand-bound R state hemoglobins are associated with a decrease in the frequency of the iron-proximal histidine stretching mode in the corresponding transient species. Such effects can originate from changes in either quaternary state or solution conditions as well as species-specific variations in protein structure. These results indicate that modulation of the Fe-His linkage could be a general mechanism for regulating ligand binding properties in hemoglobin. A direct connection between ligand binding and the Fe-His bond is suggested from our finding that the structural parameter(s) regulating the barrier height for germinate recombination is likely to be the same as the one(s) modulating the frequency of the Fe-His stretching mode in the transient deoxy species. Based on the recent conclusion that the variation in the tilt of histidine with respect to the heme plane is primarily responsible for the spectrum of frequencies observed for the Fe-His stretching mode we have examined a model in which protein regulation of binding occurs via a protein-induced change in the histidine tilt.


Subtypes of primary angiitis of the CNS identified by MRI patterns reflect the size of affected vessels.
To describe patterns of diagnostic findings, and identify subgroups of primary angiitis of the central nervous system (PACNS). We retrospectively analysed 31 patients with PACNS. Cases were selected by predetermined diagnostic criteria and stratified into biopsy-proven and imaging-based PACNS. We compared clinical characteristics, cerebrospinal fluid (CSF) findings and imaging results including high-resolution vessel wall MRI between groups. There were 31 cases of PACNS (mean age 45.6 years, 58.1% female), of whom 17 (55%) were biopsy-proven, 14 (45%) were based on imaging findings. Patients with a positive biopsy had fewer infarcts (29.4% vs 85.7%, p=0.003), were more likely to have meningeal and parenchymal contrast enhancement (76.5% vs 28.6%, p=0.012), were less likely to have abnormal MR angiography (11.8% vs 100%, p<0.001) and did not show vessel wall enhancement at the time of diagnosis (0% vs 76.9%, p<0.001). In contrast, patients with imaging-based diagnosis showed more frequently multiple infarcts and vessel abnormalities, with vessel wall enhancement in most of the cases. Clinical characteristics and CSF analysis did not reveal marked differences between groups. Multi-parametric MRI distinguishes two subtypes of PACNS that most likely differ concerning the affected vessel size. Biopsy-proven PACNS primarily involves smaller vessels beyond the resolution of vascular imaging, while imaging-based PACNS affects predominantly medium-sized vessels leading to false-negative biopsy results. Using distinct MRI patterns may be helpful for selecting patients for appropriate invasive diagnostic modalities.


The potential contribution of impaired brain glucose metabolism to congenital Zika syndrome.
The Zika virus (ZIKV) became a major worldwide public concern in 2015 due to the congenital syndrome which presents the highest risk during the first trimester of pregnancy and includes microcephaly and eye malformations. Several cellular, genetic and molecular studies have shown alterations in metabolic pathways, endoplasmic reticulum (ER) stress, immunity and dysregulation of RNA and energy metabolism both in vivo and in vitro. Here we summarise the main metabolic complications, with a particular focus on the possibility that brain energy metabolism is altered following ZIKV infection, contributing to developmental abnormalities. Brain energetic failure has been implicated in neurological conditions such as autism disorder and epilepsy, as well as in metabolic diseases with severe neurodevelopmental complications such as Glut-1 deficiency syndrome. Therefore, these energetic alterations are of wide-ranging interest as they might be directly implicated in congenital ZIKV syndrome. Data showing increased glycolysis during ZIKV infection, presumably required for viral replication, might support the idea that the virus can cause energetic stress in the developing brain cells. Consequences may include neuroinflammation, cell cycle dysregulation and cell death. Ketone bodies are non-glycolytic brain fuels that are produced during neonatal life, starvation or fasting, ingestion of high-fat low-carbohydrate diets, and following supplementation with ketone esters. We propose that dietary ketones might alter the course of the disease and could even provide some degree of prevention of ZIKV-associated abnormalities and potentially related neurological conditions characterised by brain glucose impairment.


Quantitative in situ and real-time monitoring of mechanochemical reactions.
An experimental technique for in situ and real-time monitoring of mechanochemical reactions in a shaker ball mill was recently described, which utilises highly penetrating X-ray radiation available at the ID15B beamline of the European Synchrotron Radiation Facility. Herein, we describe the first attempts to perform such reaction monitoring in a quantitative fashion, by introducing an internal X-ray diffraction standard. The use of silicon as an internal standard resolved the issue with variations of the amount of the sample in the X-ray beam due to the non-uniform distribution of the sample in the reaction jar and allowed, via Rietveld analysis, the first quantitative estimate of the amorphous phase content in a mechanochemical reaction as it is being milled. We also highlight problems associated with the non-ideal mixing of the reaction mixture.


New insights into cellular prion protein (PrPc) functions: the "ying and yang" of a relevant protein.
The conversion of cellular prion protein (PrP(c)), a GPI-anchored protein, into a protease-K-resistant and infective form (generally termed PrP(sc)) is mainly responsible for Transmissible Spongiform Encephalopathies (TSEs), characterized by neuronal degeneration and progressive loss of basic brain functions. Although PrP(c) is expressed by a wide range of tissues throughout the body, the complete repertoire of its functions has not been fully determined. Recent studies have confirmed its participation in basic physiological processes such as cell proliferation and the regulation of cellular homeostasis. Other studies indicate that PrP(c) interacts with several molecules to activate signaling cascades with a high number of cellular effects. To determine PrP(c) functions, transgenic mouse models have been generated in the last decade. In particular, mice lacking specific domains of the PrP(c) protein have revealed the contribution of these domains to neurodegenerative processes. A dual role of PrP(c) has been shown, since most authors report protective roles for this protein while others describe pro-apoptotic functions. In this review, we summarize new findings on PrP(c) functions, especially those related to neural degeneration and cell signaling.


Transcriptional responses to sucrose mimic the plant-associated life style of the plant growth promoting endophyte Enterobacter sp. 638.
Growth in sucrose medium was previously found to trigger the expression of functions involved in the plant associated life style of the endophytic bacterium Enterobacter sp. 638. Therefore, comparative transcriptome analysis between cultures grown in sucrose or lactate medium was used to gain insights in the expression levels of bacterial functions involved in the endophytic life style of strain 638. Growth on sucrose as a carbon source resulted in major changes in cell physiology, including a shift from a planktonic life style to the formation of bacterial aggregates. This shift was accompanied by a decrease in transcription of genes involved in motility (e.g., flagella biosynthesis) and an increase in the transcription of genes involved in colonization, adhesion and biofilm formation. The transcription levels of functions previously suggested as being involved in endophytic behavior and functions responsible for plant growth promoting properties, including the synthesis of indole-acetic acid, acetoin and 2,3-butanediol, also increased significantly for cultures grown in sucrose medium. Interestingly, despite an abundance of essential nutrients transcription levels of functions related to uptake and processing of nitrogen and iron became increased for cultures grown on sucrose as sole carbon source. Transcriptome data were also used to analyze putative regulatory relationships. In addition to the small RNA csrABCD regulon, which seems to play a role in the physiological adaptation and possibly the shift between free-living and plant-associated endophytic life style of Enterobacter sp. 638, our results also pointed to the involvement of rcsAB in controlling responses by Enterobacter sp. 638 to a plant-associated life style. Targeted mutagenesis was used to confirm this role and showed that compared to wild-type Enterobacter sp. 638 a ΔrcsB mutant was affected in its plant growth promoting ability.


Identification of bottleneck enzymes with negative dynamic sensitivities: ethanol fermentation systems as case studies.
Dynamic logarithmic gains (normalised time-varying sensitivities) or their products with metabolite concentrations, called an instantaneous bottleneck ranking indicator (instantaneous BR indicator), can express the time courses of bottleneck enzyme ranking. The integrated value of the instantaneous BR indicator, called an overall BR indicator, provides a ranking of bottleneck enzymes over the entire reaction period of time. Owing to the property of the logarithmic gains, these BR indicators are allowed to include both positive and negative values and in turn raise several questions on the identification of a bottleneck enzyme. To eliminate these questions, the present work studies the identification of bottleneck enzymes in four ethanol fermentation systems using suspended and immobilised cells at pH 4.5 and 5.5 as case studies. These systems consist of both intracellular reactions without cell growth (intracellular reaction systems) and those with cell growth (cell growth systems). The analyses of the intracellular reaction systems reveal that both the suspended and immobilised cell systems at pH 5.5 are not robust. In the cell growth systems, therefore, the analyses are conducted only for the suspended and immobilised cell models at pH 4.5. The enzyme glyceraldehyde 3-phosphate dehydrogenase (gapd) initially has the largest logarithmic gains under all conditions, whereas the system becomes insensitive to it over time. Interestingly, the instantaneous BR indicator for gapd is negatively large. Also, other indicators change in positive and/or negative regions. To deal with these values of different signs, all the indicators are integrated to obtain and rank the overall BR indicators in the order of the magnitudes of their absolute values. The result shows that the ranking of enzymes are completely identical to that of the final ethanol concentrations obtained by finitely increasing or decreasing the relevant enzyme activities in accordance with the signs of overall BR indicators. This result indicates that the enzymes must be ranked in the order of the magnitudes of their absolute values to determine a bottleneck enzyme. As a result, the analyses of the intracellular reaction systems reveal that the bottleneck enzyme is switched from a membrane-bound glucose transporter enzyme to phosphofructokinase when the cell-retaining condition is changed from suspension to immobilisation. In the cell growth systems, gapd is identified as the most likely bottleneck enzyme, suggesting that the gapd activity must be decreased to increase the ethanol productivity.


c-RAF Ablation Induces Regression of Advanced Kras/Trp53 Mutant Lung Adenocarcinomas by a Mechanism Independent of MAPK Signaling.
A quarter of all solid tumors harbor KRAS oncogenes. Yet, no selective drugs have been approved to treat these malignancies. Genetic interrogation of the MAPK pathway revealed that systemic ablation of MEK or ERK kinases in adult mice prevent tumor development but are unacceptably toxic. Here, we demonstrate that ablation of c-RAF expression in advanced tumors driven by KrasG12V/Trp53 mutations leads to significant tumor regression with no detectable appearance of resistance mechanisms. Tumor regression results from massive apoptosis. Importantly, systemic abrogation of c-RAF expression does not inhibit canonical MAPK signaling, hence, resulting in limited toxicities. These results are of significant relevance for the design of therapeutic strategies to treat K-RAS mutant cancers.


RAMBO-K: Rapid and Sensitive Removal of Background Sequences from Next Generation Sequencing Data.
The assembly of viral or endosymbiont genomes from Next Generation Sequencing (NGS) data is often hampered by the predominant abundance of reads originating from the host organism. These reads increase the memory and CPU time usage of the assembler and can lead to misassemblies. We developed RAMBO-K (Read Assignment Method Based On K-mers), a tool which allows rapid and sensitive removal of unwanted host sequences from NGS datasets. Reaching a speed of 10 Megabases/s on 4 CPU cores and a standard hard drive, RAMBO-K is faster than any tool we tested, while showing a consistently high sensitivity and specificity across different datasets. RAMBO-K rapidly and reliably separates reads from different species without data preprocessing. It is suitable as a straightforward standard solution for workflows dealing with mixed datasets. Binaries and source code (java and python) are available from http://sourceforge.net/projects/rambok/.


Does Usage of an eHealth Intervention Reduce the Risk of Excessive Gestational Weight Gain? Secondary Analysis From a Randomized Controlled Trial.
Excessive gestational weight gain (GWG) contributes to the development of obesity in mother and child. Internet-based interventions have the potential for delivering innovative and interactive options for prevention of excessive GWG to large numbers of people. The objective of this study was to create a novel measure of Internet-based intervention usage patterns and examine whether usage of an Internet-based intervention is associated with reduced risk of excessive GWG. The website featured blogs, local resources, articles, frequently asked questions (FAQs), and events that were available to women in both the intervention and control arm. Weekly reminders to use the website and to highlight new content were emailed to participants in both arms. Only intervention arm participants had access to the weight gain tracker and diet and physical activity goal-setting tools. A total of 1335 (898 intervention and 437 control) relatively diverse and healthy pregnant women were randomly assigned to the intervention arm or control arm. Usage patterns were examined for both intervention and control arm participants using latent class analysis. Regression analyses were used to estimate the association between usage patterns and three GWG outcomes: excessive total GWG, excessive GWG rate, and GWG. Five usage patterns best characterized the usage of the intervention by intervention arm participants. Three usage patterns best characterized control arm participants' usage. Control arm usage patterns were not associated with excessive GWG, whereas intervention arm usage patterns were associated with excessive GWG. The control and intervention arm usage pattern characterization is a unique methodological contribution to process evaluations for self-directed Internet-based interventions. In the intervention arm some usage patterns were associated with GWG outcomes. ClinicalTrials.gov; Clinical Trials Number: NCT01331564; https://clinicaltrials.gov/ct2/show/NCT01331564 (Archived by WebCite at http://www.webcitation/6nI9LuX9w).