Role of homology in site-specific recombination of bacteriophage lambda: evidence against joining of cohesive ends.
Bacteriophage lambda integration and excision take place at specific loci called attachment sites. Earlier work has shown that efficient recombination requires the identical sequence to be present in both attachment sites throughout the seven-base-pair region between the points of strand exchange. A plausible model for the role of homology postulates that Int, the site-specific recombinase, makes double-strand breaks at attachment sites such that each broken end has a short single-strand protrusion. Recombination would then depend upon the capacity of these protrusions to form Watson-Crick helices--i.e., to anneal--a process that might require perfect complementarity between the cohesive ends. To test this model, we have studied Int-promoted crosses in which one attachment site is a heteroduplex. Specifically, we constructed sites in which the seven-base-pair region between the points of strand exchange contains one or more noncomplementary pairs. The double-strand break and annealing mechanism predicts that crosses with these heteroduplex sites should yield one completed recombinant and one broken site. We find that such nonreciprocal recombination is uncommon and that the typical outcome of crosses involving a heteroduplex site is a reciprocal recombinant in which both products are resealed. Moreover, the occasional appearance of nonreciprocal products can be explained by our finding that Int can cleave heteroduplex attachment sites after recombination is completed. Taken together, our data strongly indicate that bacteriophage lambda recombination does not proceed by the homology-dependent annealing of cohesive ends; acceptable alternatives for the role of homology are discussed.


Severe community-acquired pneumonia requiring intensive care: a study of 80 cases from Singapore.
Severe community-acquired pneumonia is a major cause of mortality and morbidity worldwide. This study looked at the clinical characteristics of these patients admitted to a Singaporean community hospital and the prognostic impact of age, bacteraemia and logistic organ dysfunction score (LODS) on intensive care unit mortality. Retrospective analysis of 80 severe community-acquired pneumonia patients admitted to the intensive care unit over a 20-month period was conducted. The Mann-Whitney U and chi-square tests were used for statistical analysis and a p-value of less than 0.5 was considered as significant. There were 55 male and 25 female patients, with a median age of 62 years. The median LODS was 5. The intensive care unit mortality was 30 percent. The median LODS of intensive care unit survivors was 5 and of non-survivors, 8. The overall hospital mortality was 37.5 percent. A microbiological aetiology was identified in 38 percent of patients. Three of four patients with melioidosis died. Between intensive care unit survivors and non-survivors, there was a significant difference in the LODS, but no significant difference in the age and incidence of bacteraemia. Severe community-acquired pneumonia is a highly fatal disease which requires early initiation of appropriate empirical antibiotic therapy, which should include coverage for melioidosis in the local context. The microbiological workup should include testing for tuberculosis. The LODS system may be an appropriate tool in estimating the severity of illness.


STAT5 is required for long-term maintenance of normal and leukemic human stem/progenitor cells.
The transcription factor STAT5 fulfills a distinct role in the hematopoietic system, but its precise role in primitive human hematopoietic cells remains to be elucidated. Therefore, we performed STAT5 RNAi in sorted cord blood (CB) and acute myeloid leukemia (AML) CD34+ cells by lentiviral transduction and investigated effects of STAT5 downmodulation on the normal stem/progenitor cell compartment and the leukemic counterpart. STAT5 RNAi cells displayed growth impairment, without affecting their differentiation in CB and AML cultures on MS5 stroma. In CB, limiting-dilution assays demonstrated a 3.9-fold reduction in progenitor numbers. Stem cells were enumerated in long-term culture-initiating cell (LTC-IC) assays, and the average LTC-IC frequency was 3.25-fold reduced from 0.13% to 0.04% by STAT5 down-regulation. Single-cell sorting experiments of CB CD34+/CD38(-) cells demonstrated a 2-fold reduced cytokine-driven expansion, with a subsequent 2.3-fold reduction of progenitors. In sorted CD34+ AML cells with constitutive STAT5 phosphorylation (5/8), STAT5 RNAi demonstrated a reduction in cell number (72% +/- 17%) and a decreased expansion (17 +/- 15 vs 80 +/- 58 in control cultures) at week 6 on MS5 stroma. Together, our data indicate that STAT5 expression is required for the maintenance and expansion of primitive hematopoietic stem and progenitor cells, both in normal as well as leukemic hematopoiesis.


Speech overlap in Japanese mother-child conversations.
This study aimed to clarify the development of conversational style in Japanese mother-child interactions. We focused on the frequency of speech overlap as an index of Japanese conversational style, with particular attention to ne, a particle produced by the speaker, and to backchannels, such as 'uh-huh', produced by the listener that support sympathetic conversation. The results of longitudinal observations of two Japanese mother-child dyads from approximately 0;11 to 3;3 suggest that an adultlike conversational style with frequent overlaps emerges in Japanese child-directed speech around the two-word utterance period, and a child's development of ne use is closely related to this shift.


Improving public health by tackling climate change.
Across the world, climate change is now responsible for substantial mortality and morbidity, through direct effects on health and also by threatening the determinants of health. This commentary argues that adaptation policies to enhance resilience to adverse climate events are important, but must be coupled with determined action to reduce greenhouse gas emissions. The prize is synergy: many such policies, for example concerning food, travel and community engagement, can simultaneously improve physical and mental health.This is a commentary on http://www.ijhpr.org/content/2/1/23.


Central and East European migrant men who have sex with men in London: a comparison of recruitment methods.
Following the expansion of the European Union, there has been a large influx of Central and East European (CEE) migrants to the UK. CEE men who have sex with men (MSM) represent a small minority within this population that are none-the-less important to capture in sexual health research among the CEE migrant community. This paper examines the feasibility of recruiting CEE MSM for a survey of sexual behaviour in London using respondent driven sampling (RDS), via gay websites and in GUM clinics. We sought CEE MSM to start RDS chain referral among GUM clinic attendees, our personal contacts and at gay events and venues in central London. We recruited CEE MSM (n = 485) via two popular websites for gay men in Britain (March-May 2009) and at two central London GUM clinics (n = 51) (July 2008-March 2009). We found seventeen men who knew other CEE MSM in London and agreed to recruit contacts into the study. These men recruited only three men into the study, none of whom recruited any further respondents, and RDS was abandoned after 7 months (July 2008-January 2009). Half of the men that we approached to participate in RDS did not know any other CEE MSM in London. Men who agreed to recruit contacts for RDS were rather more likely to have been in the UK for more than one year (94.1% vs 70.0%, p = 0.052). Men recruited through gay websites and from GUM clinics were similar. The Internet was the most successful method for collecting data on sexual risk behaviour among CEE MSM in London. CEE MSM in London were not well networked. RDS may also have failed because they did not fully understand the procedure and/or the financial incentive was not sufficient motivation to take part.


Efficient Cs+-Sr2+ Separation over a Microporous Silver Selenidostannate Synthesized in Deep Eutectic Solvent.
Efficient Cs+-Sr2+ separation, highly desirable for radionuclide recovery in medical and industrial applications, was achieved by the ion exchange technique over a novel microporous silver selenidostannate, [NH3CH3]0.5[NH2(CH3)2]0.25Ag1.25SnSe3 (AgSnSe-1). This material was synthesized in deep eutectic solvent (DES), where the alkylammonium cations play significant structure-directing roles in the construction of micropores that allow for selective ion exchange toward Cs+ against Sr2+. The much greater KdCs (1.06 × 104 mL g-1) over KdSr (87.7 mL g-1) contributes to an outstanding separation factor SFCs/Sr of ∼121.4 that is top-ranked among inorganic materials. An ion exchange column filled with AgSnSe-1 exhibits a remarkable separation effect for 10 000 bed volumes of continuous flow, with removal rates of ∼99.9% and ∼0 ± 5.5% for Cs+ and Sr2+, respectively. AgSnSe-1 exhibits excellent β and γ radiation resistances and a chemical stability over a broad pH range of 1-12. The Se leaching level below the safe guideline value for drinking water highlights the environmental-friendly nature of AgSnSe-1. The high Cs+ exchange performance is almost unaffected by Na+, Mg2+, and Ca2+ cations. The Cs+-laden product AgSnSe-1Cs can be facilely eluted for recycling use, highlighting the great potential of open framework metal selenides in nuclear waste treatment and renewable energy utilization.


[Dietetic care for patients after maxillofacial trauma].
Patients suffering from maxillofacial trauma and subsequent intermaxillary fixation are temporary at risk of severe malnutrition and substantial weight loss. Eventually, these consequences may lead to impaired fracture healing and wound healing, and thus to deteriorated overall functional recovery. Adequate evaluation of several specific risk factors warrants the opportunity to stratify individual patients to be at either high or low risk for nutrition related health problems. Customary dietary assessment and counselling can be provided when necessary.


Incidence of venous thromboembolism and bleeding events in patients with lower extremity orthopedic surgery: a retrospective analysis of a Japanese healthcare database.
Orthopedic surgeries of lower extremities such as total knee arthroplasty (TKA), total hip arthroplasty (THA), and hip fracture surgery (HFS) are widely considered to carry a high risk of developing deep venous thrombosis (DVT) and pulmonary thromboembolism (PTE). Growing attention to epidemiological studies using a healthcare database led us to quantify the risks using a Japanese database to reveal recent medical care for such events. The study comprised 36,947 patients who had undergone orthopedic surgeries of the lower extremities and whose medical information from April 2008 to September 2013 was available. The source population of the database was derived from 100 acute-care hospitals with Diagnosis Procedure Combination. The events were defined by diagnosis, medication, imaging, and laboratory tests. A breakdown of patients who underwent orthopedic surgeries by type of surgery showed 13.6% for TKA, 10.4% for THA, 56.8% for HFS, 1.5% for rupture of Achilles tendon, and 18.0% for simple fracture of lower extremities. The incidence for DVT, PTE, and bleeding events by type of surgery was 1.3, 0.2, and 1.0% for TKA; 0.9, 0.2, and 1.1% for THA; and 0.4, 0.1, and 1.8% for HFS, respectively. The population for risk factor analysis consisted of patients with similar background factors who underwent TKA, THA, or HFS. The statistically significant risk factors for PTE analyzed by the backward elimination procedure in a multivariate model were female sex, history of venous thromboembolism (VTE), thrombophilia, and varicose veins of lower extremity. The incidence of DVT, PTE, and bleeding events and the risk factors for DVT and PTE in patients by type of orthopedic surgeries of lower extremities found in our study are considered to be rational as they reflect evidence from real-world cases. University Hospital Medical Information Network Clinical Trials Registry UMIN000012667.


Male and female employment in the textile industry in relation to miscarriage and preterm delivery.
To address potential reproductive hazards in textile manufacturing, we conducted a community-based case-control study in central North Carolina. Miscarriage cases were identified from medical records (280 interviewed cases): preterm delivery cases and term, normal birth weight controls (454 and 605, respectively) were identified from area hospitals. Exposures were based on job title, an interview concerning textile-related exposures, expert imputation of exposure based on job titles and interviews, and self-reported exposures by women. Relative to women and men working in nonhazardous occupations, workers in the textile industry were not at increased risk of miscarriage or preterm delivery, with the possible exception of preterm delivery among women and men employed in sectors other than knitting and yarn mills and men employed in yarn mills. Inferred exposures to specific agents were also not associated with adverse pregnancy outcome. Subject to uncertainty in exposure assessment and nonresponse, these data indicate an absence of adverse effects of the textile workplace environment on these pregnancy outcomes.


A comparison of visits and practices of nurse-midwives and obstetrician-gynecologists in ambulatory care settings.
With more than 5 million patient visits annually, certified nurse-midwives (CNMs) substantially contribute to women's health care in the United States. The objective of this study was to describe ambulatory visits and practices of CNMs, and compare them with those of obstetrician-gynecologists (OB/GYNs). Sources of population-based data used to compare characteristics of provider visits were three national surveys of CNMs and two National Ambulatory Medical Care Surveys of physicians. When a subset of 4,305 visits to CNMs in 1991 and 1992 were compared to 5,473 visits to OB/GYNs in similar office-based ambulatory care settings in 1989 and 1990, it was found that a larger proportion of CNM visits were made by women who were publicly insured and below age 25. The majority of visits to CNMs were for maternity care; the majority of visits to OB/GYNs were for gynecologic and/or family planning concerns. Face-to-face visit time was longer for CNMs, and involved more client education or counseling. This population-based comparison suggests that CNMs and OB/GYNs provide ambulatory care for women with diverse demographic characteristics and differing clinical service needs. Enhancing collaborative practice could improve health care access for women, which would be especially beneficial for those who are underserved and vulnerable.


Comparison of the rolling circle amplification and ligase-dependent reaction methods for the identification of opportunistic Exophiala species.
We developed two ligase-dependent probe amplification assays based on rolling circle amplification (RCA) and the ligase-dependent reaction (LDR) to differentiate species of Exophiala targeting the rDNA internal transcribed spacer region. We focused on Exophiala dermatitidis and E. phaeomuriformis, two opportunistic inhabitants of indoor wet cells, and further detected E. heteromorpha, E. xenobiotica, and E. crusticola; 57 reference isolates representing the five species were tested. Depending on the RCA probes used, the sensitivity was 100%, and the specificity ranged from 3.7% to 88.6% (median: 46.1%). In contrast, the sensitivity and specificity of the LDR probes targeting the same isolates were 88.6-100% (median: 95.8%) and 95.4-100% (median: 97.7%), respectively. We analyzed 198 additional environmental isolates representing the same Exophiala species. Overall, the sensitivity and specificity of LDR ranged from 89.7% to 100% (median: 94.1%) and from 93.9% to 100% (median: 96.9%), respectively. The assessment of performance and validation of LDR probes using SYBR Green quantitative polymerase chain reaction revealed high reproducibility and an acceptable range limit, in line with the guidelines of the European Network of GMO Laboratories. In conclusion, the LDR assay was more reliable and less expensive than RCA for species-level identification of Exophiala isolates.


HIV neuropathy: insights in the pathology of HIV peripheral nerve disease.
HIV-associated neuropathies (HIV-N) have become the most frequent neurological disorder associated with HIV infection. The most common forms of HIV-N are the distal sensory polyneuropathy (DSP) and antiretroviral toxic neuropathies (ATN), disorders characterized mostly by sensory symptoms that include spontaneous or evoked pain that follow a subacute or chronic course. The main pathological features that characterize DSP and ATN include "dying back" axonal degeneration of long axons in distal regions, loss of unmyelinated fibers, and variable degree of macrophage infiltration in peripheral nerves and dorsal root ganglia. Marked activation of macrophages as well as the effect of pro-inflammatory cytokines appear to be the main immunopathogenic factors in DSP. Interference with DNA synthesis and mitochondrial abnormalities produced by nucleoside antiretrovirals have been hypothesized as pathogenic factors involved in ATN. The use of skin biopsy has become a useful tool in the evaluation of HIV-N. Reduction in fiber density, increased frequency of fiber varicosities and fiber fragmentation are prominent features of skin biopsies from patients with HIV-N. Other forms of HIV-N include acute or chronic inflammatory polyneuropathies, uncommon disorders that may ocur during seroconversion or early stages of HIV infection. Opportunisitic infections, mostly associated with cytomegalovirus or herpes zoster virus infection occur in late stages of AIDS and produce characteristic clinical features such as mononeuritis multiple or radiculopathies.


The targeted inactivation of TRβ gene in thyroid follicular cells suggests a new mechanism of regulation of thyroid hormone production.
Thyroid epithelial cells, or thyrocytes, express functional thyroid hormone receptors but no precise role has yet been assigned to either TRα or TRβ in the thyroid gland. In this study, we analyzed the impact of inactivating the TRβ gene in the thyroid of mice. First, we generated a mouse line named Thyr-Cre, expressing the Cre recombinase under the control of the thyroglobulin gene promoter, which led to a complete recombination of floxed genes in thyrocytes. Thyr-Cre mice were then crossed with TRβ floxed mice (TRβ(flox/flox)) to obtain a thyrocyte-selective deletion of TRβ. Thyr-TRβ(-/-) mice were characterized by a decrease in the size and functional activity of the thyroid gland. These alterations were associated with a decrease in plasma TSH concentration. Surprisingly, Thyr-TRβ(-/-) displayed elevated serum T(4) and rT(3) concentrations with no significant change in serum T(3) levels. Their intrathyroidal free T(4) and rT(3) contents were also elevated, whereas the ratio of serum T(4) to thyroid free T(4) was decreased by comparison with wild-type littermates. Also, within the thyroid, deiodinases D1 and D2 were reduced as well as the expression levels of genes encoding monocarboxylate transporters (Mct8 and Mct10). Such a decrease in intrathyroidal deiodination of T(4) and in the expression of genes encoding thyroid hormone transporters may contribute to the primary overproduction of T(4) observed in Thyr-TRβ(-/-) mice. In conclusion, these data show that the control of thyroid hormone production involves not only TRβ-dependent mechanisms acting at the level of hypothalamus and pituitary but also TRβ-dependent mechanisms acting at the thyroid level.


Electrical and optical percolations of polystyrene latex-multiwalled carbon nanotube composites.
Electrical conductivity and optical transmittance properties of polystyrene (PS)-multiwalled carbon nanotube (MWCNT) composite films were investigated. Composite films were prepared by mixing of various mass fractions of MWCNT in PS-water dispersions. After water evaporates, powder composite films were annealed at 175 degrees C above the glass transition of PS for 20 min. Photon transmission and two point probe resistivity techniques were employed to determine the variations of the optical and the electrical properties of composites. Transmitted light intensity, I(tr) and surface resistivity, R(s) were monitored as a function of MWCNT mass fraction (M). It was observed that, both the surface resistivity and the optical transparency were decreased by increasing the amount of MWCNT added to the polymeric system. Conductivity and optical results were interpreted according to the classical and site percolation theory, respectively. The electrical (sigma) and the optical (op) percolation threshold values and critical exponents were calculated as M(sigma)=1.8 wt.%, M(op)=0-0.13 wt.% and beta(sigma)=2.25, beta(op)=0.32, respectively.


Organic matrix synthesis in the scleractinian coral stylophora pistillata: role in biomineralization and potential target of the organotin tributyltin
The kinetics of organic matrix biosynthesis and incorporation into scleractinian coral skeleton was studied using microcolonies of Stylophora pistillata. [14C]Aspartic acid was used to label the organic matrix since this acidic amino acid can represent up to 50 mol % of organic matrix proteins. External aspartate was rapidly incorporated into tissue protein without any detectable lag phase, suggesting either a small intracellular pool of aspartic acid or a pool with a fast turn-over rate. The incorporation of 14C-labelled macromolecules into the skeleton was linear over time, after an initial delay of 20 min. Rates of calcification, measured by the incorporation of 45Ca into the skeleton, and of organic matrix biosynthesis and incorporation into the skeleton were constant. Inhibition of calcification by the Ca2+ channel inhibitor verapamil reduced the incorporation of organic matrix proteins into the skeleton. Similarly, organic matrix incorporation into the skeleton, but not protein synthesis for incorporation into the tissue compartment, was dependent on the state of polymerization of both actin and tubulin, as shown by the sensitivity of this process to cytochalasin B and colchicin. These drugs may inhibit exocytosis of organic matrix proteins into the subcalicoblastic space. Finally, inhibition of protein synthesis by emetin or cycloheximide and inhibition of N-glycosylation by tunicamycin reduced both the incorporation of macromolecules into the skeleton and the rate of calcification. This suggests that organic matrix biosynthesis and its migration towards the site of calcification may be a prerequisite step in the calcification process. On the basis of these results, we investigated the effects of tributyltin (TBT), a component of antifouling painting known to interfere with biomineralization processes. Our results have shown that this xenobiotic significantly inhibits protein synthesis and the subsequent incorporation of protein into coral skeleton. This effect was correlated with a reduction in the rate of calcification. Protein synthesis was shown to be the parameter most sensitive to TBT (IC50=0.2 micromol l-1), followed by aspartic acid uptake by coral tissue (IC50=0.6 micromol l-1), skeletogenesis (IC50=3 micromol l-1) and Ca2+ uptake by coral tissue (IC50=20 micromol l-1). These results suggest that the mode of action of TBT on calcification may be the inhibition of organic matrix biosynthesis.


Duration of antibiotic therapy for bacteremia: a systematic review and meta-analysis.
The optimal duration of antibiotic therapy for bloodstream infections is unknown. Shorter durations of therapy have been demonstrated to be as effective as longer durations for many common infections; similar findings in bacteremia could enable hospitals to reduce antibiotic utilization, adverse events, resistance and costs. A search of the MEDLINE, EMBASE and COCHRANE databases was conducted for the years 1947-2010. Controlled trials were identified that randomized patients to shorter versus longer durations of treatment for bacteremia, or the infectious foci most commonly causing bacteremia in critically ill patients (catheter-related bloodstream infections (CRBSI), intra-abdominal infections, pneumonia, pyelonephritis and skin and soft-tissue infections (SSTI)). Twenty-four eligible trials were identified, including one trial focusing exclusively on bacteremia, zero in catheter related bloodstream infection, three in intra-abdominal infection, six in pyelonephritis, thirteen in pneumonia and one in skin and soft tissue infection. Thirteen studies reported on 227 patients with bacteremia allocated to 'shorter' or 'longer' durations of treatment. Outcome data were available for 155 bacteremic patients: neonatal bacteremia (n = 66); intra-abdominal infection (40); pyelonephritis (9); and pneumonia (40). Among bacteremic patients receiving shorter (5-7 days) versus longer (7-21 days) antibiotic therapy, no significant difference was detected with respect to rates of clinical cure (45/52 versus 47/49, risk ratio 0.88, 95% confidence interval [CI] 0.77-1.01), microbiologic cure (28/28 versus 30/32, risk ratio 1.05, 95% CI 0.91-1.21), and survival (15/17 versus 26/29, risk ratio 0.97, 95% CI 0.76-1.23). No significant differences in clinical cure, microbiologic cure and survival were detected among bacteremic patients receiving shorter versus longer duration antibiotic therapy. An adequately powered randomized trial of bacteremic patients is needed to confirm these findings.


Method for the isolation of Francisella tularensis outer membranes.
Francisella tularensis is a Gram-negative intracellular coccobacillus and the causative agent of the zoonotic disease tularemia. When compared with other bacterial pathogens, the extremely low infectious dose (<10 CFU), rapid disease progression, and high morbidity and mortality rates suggest that the virulent strains of Francisella encode for novel virulence factors. Surface-exposed molecules, namely outer membrane proteins (OMPs), have been shown to promote bacterial host cell binding, entry, intracellular survival, virulence and immune evasion. The relevance for studying OMPs is further underscored by the fact that they can serve as protective vaccines against a number of bacterial diseases. Whereas OMPs can be extracted from gram-negative bacteria through bulk membrane extraction techniques, including sonication of cells followed by centrifugation and/or detergent extraction, these preparations are often contaminated with periplasmic and/or cytoplasmic (inner) membrane (IM) contaminants. For years, the "gold standard" method for the biochemical and biophysical separation of gram-negative IM and outer membranes (OM) has been to subject bacteria to spheroplasting and osmotic lysis, followed by sucrose density gradient centrifugation. Once layered on a sucrose gradient, OMs can be separated from IMs based on the differences in buoyant densities, believed to be predicated largely on the presence of lipopolysaccharide (LPS) in the OM. Here, we describe a rigorous and optimized method to extract, enrich, and isolate F. tularensis outer membranes and their associated OMPs.


Quorum quenching Bacillus sonorensis isolated from soya sauce fermentation brine.
An N-acylhomoserine lactone (AHL)-degrading bacterial strain, L62, was isolated from a sample of fermentation brine of Chinese soya sauce by using rich medium agar supplemented with soya sauce (10% v/v). L62, a rod-shaped Gram positive bacterium with amylolytic activity, was phylogentically related to Bacillus sonorensis by 16S ribosomal DNA and rpoB sequence analyses. B. sonorensis L62 efficiently degraded N-3-oxohexanoyl homoserine lactone and N-octanoylhomoserine lactone. However, the aiiA homologue, encoding an autoinducer inactivation enzyme catalyzing the degradation of AHLs, was not detected in L62, suggesting the presence of a different AHL-degrading gene in L62. To the best of our knowledge, this is the first report of AHL-degrading B. sonorensis from soya sauce liquid state fermentation.


Tracking the ecological overshoot of the human economy.
Sustainability requires living within the regenerative capacity of the biosphere. In an attempt to measure the extent to which humanity satisfies this requirement, we use existing data to translate human demand on the environment into the area required for the production of food and other goods, together with the absorption of wastes. Our accounts indicate that human demand may well have exceeded the biosphere's regenerative capacity since the 1980s. According to this preliminary and exploratory assessment, humanity's load corresponded to 70% of the capacity of the global biosphere in 1961, and grew to 120% in 1999.


Validation of apathy evaluation scale and assessment of severity of apathy in Alzheimer's disease.
Apathy is a well-recognized symptom of Alzheimer's disease (AD). The aim of the present study was to validate the Taiwanese version of the Apathy Evaluation Scale, clinician version (AES-C) and assess the severity of apathy in patients with AD. Comprehensive evaluations were conducted in a total of 144 AD patients. This study used a cross-sectional comparative design. Data were collected from clinical interviews using the AES, the Mini-Mental Status Examination (MMSE), the Neuropsychiatric Inventory (NPI), and the Clinical Dementia Rating Scale (CDR). Internal consistency was indicated by Cronbach's alphas of 0.85; test-retest reliability was 0.89 over a period of 3 days. Criterion-related validity was supported by the fact that AES-C significantly correlated with the apathy scores of the NPI. Factor analysis indicated a three-factor structure. Convergent validity was supported by a positive correlation between the AES-C score and the anxiety score of the NPI. Discriminant validity was supported by the fact that the AES-C scores did not correlate with the depression subscale of the NPI, and the correlation between the AES-C score and the euphoria score of the NPI score was negative. Known-group validity was supported by results showing that AD patients in a moderate stage of dementia (CDR = 2) had significantly higher scores on the AES-C than patients with mild-stage dementia (CDR = 1). The AES-C is a reliable and valid instrument for measuring symptoms of apathy among AD patients in Taiwan.


Differential Expression Analysis Based on Ensemble Strategy on miRNA Profiles of Kidney Clear Cell Carcinoma.
Kidney clear cell carcinoma (KIRC) is the most common type of kidney cancer, accounting for approximately 60-85% of all the kidney cancers. However, there are few options available for early treatment. Therefore, it is extremely important to identify biomarkers and study therapeutic targets for KIRC. Since there are few studies on KIRC, we used a data-driven approach to identify differential genes. Here, we used miRNA gene expression profile data from the TCGA database species of KIRC and proposed a machine learning-based approach to quantify the importance score of each gene. Then, an ensemble method was utilized to find the optimal subset of genes used to predict KIRC by clustering. The most genetic subset was then used to classify and predict KIRC. Differential genes were screened by several traditional differential analysis methods, and the selected gene subset showed a better performance. Independent testing sets from the GEO database were used to verify the effectiveness of the optimal subset of genes. Besides, cross-validation was made to verify the effectiveness of the approach. Finally, important genes, such as miR-140 and miR-210, were found to be involved in the biochemical processes of KIRC, which also proved the effectiveness of our approach.


A cross-species framework to identify vocal learning abilities in mammals.
Vocal production learning (VPL) is the experience-driven ability to produce novel vocal signals through imitation or modification of existing vocalizations. A parallel strand of research investigates acoustic allometry, namely how information about body size is conveyed by acoustic signals. Recently, we proposed that deviation from acoustic allometry principles as a result of sexual selection may have been an intermediate step towards the evolution of vocal learning abilities in mammals. Adopting a more hypothesis-neutral stance, here we perform phylogenetic regressions and other analyses further testing a potential link between VPL and being an allometric outlier. We find that multiple species belonging to VPL clades deviate from allometric scaling but in the opposite direction to that expected from size exaggeration mechanisms. In other words, our correlational approach finds an association between VPL and being an allometric outlier. However, the direction of this association, contra our original hypothesis, may indicate that VPL did not necessarily emerge via sexual selection for size exaggeration: VPL clades show higher vocalization frequencies than expected. In addition, our approach allows us to identify species with potential for VPL abilities: we hypothesize that those outliers from acoustic allometry lying above the regression line may be VPL species. Our results may help better understand the cross-species diversity, variability and aetiology of VPL, which among other things is a key underpinning of speech in our species. This article is part of the theme issue 'Voice modulation: from origin and mechanism to social impact (Part II)'.


Vocational rehabilitation improves cognition and negative symptoms in schizophrenia.
Several studies in schizophrenia found a positive association between cognitive performance and work status, and it has been reported that good cognitive performance at the outset does predict the success of vocational interventions. However little has been done to investigate whether vocational interventions itself benefit cognitive performance. To test this hypothesis we performed a randomized, placebo-controlled trial to investigate in remitted schizophrenic patients the effect of a 6-months vocational rehabilitation program on cognitive performance. We recruited 112 remitted and clinically stable schizophrenic patients who aimed to enter a vocational rehabilitation program. From these, 57 immediately entered a 6-months vocational rehabilitation program, whereas the remaining 55 were allocated to a waiting-list; the latter formed our control group, which received during the 6 months out-clinic follow-up treatment. Before and after the 6-months period we assessed changes in cognitive performance through a neuropsychological test battery, as well as changes in the psychopathological status and in quality of life. We found that vocational rehabilitation significantly improved patients' performance in cognitive measures that assess executive functions (concept formation, shifting ability, flexibility, inhibitory control, and judgment and critics abilities). Moreover, after 6 months the vocational group improved significantly in the negative symptoms and in quality of life, as compared to controls. Together with results from the literature, our findings reinforce the notion that the inclusion of vocational interventions may enhance the effectiveness of therapeutic strategies for schizophrenia patients.


Evolutionary models demonstrate rapid and adaptive diversification of Australo-Papuan pythons.
Lineages may diversify when they encounter available ecological niches. Adaptive divergence by ecological opportunity often appears to follow the invasion of a new environment with open ecological space. This evolutionary process is hypothesized to explain the explosive diversification of numerous Australian vertebrate groups following the collision of the Eurasian and Australian plates 25 Mya. One of these groups is the pythons, which demonstrate their greatest phenotypic and ecological diversity in Australo-Papua (Australia and New Guinea). Here, using an updated and near complete time-calibrated phylogenomic hypothesis of the group, we show that following invasion of this region, pythons experienced a sudden burst of speciation rates coupled with multiple instances of accelerated phenotypic evolution in head and body shape and body size. These results are consistent with adaptive radiation theory with an initial rapid niche-filling phase and later slow-down approaching niche saturation. We discuss these findings in the context of other Australo-Papuan adaptive radiations and the importance of incorporating adaptive diversification systems that are not extraordinarily species-rich but ecomorphologically diverse to understand how biodiversity is generated.


Prenatal diagnosis of dilated cavum septum pellucidum et vergae.
The cavum septum pellucidum is a fluid-filled cavity located between the membranes of the septi pellucidi, which should always be visualized sonographically in normal fetuses between 18 and 37 weeks gestation and in approximately 50% of term infants. The cavum vergae, a cavity within the septum pellucidum, is located posterior to an arbitrary vertical plane formed by the columns of the fornix. We present a case in which markedly dilated cavum septum pellucidum et vergae noted at 29 weeks gestation was associated with midgut malrotation and volvulus, and review the literature pertaining to this unusual prenatal ultrasonographic diagnosis.


Long Noncoding RNA TP73-AS1 Targets MicroRNA-329-3p to Regulate Expression of the SMAD2 Gene in Human Cervical Cancer Tissue and Cell Lines.
BACKGROUND Worldwide, mortality from cervical cancer in women remains high. This study aimed to investigate the expression of long noncoding RNA (lncRNA) TP73-AS1, microRNA-329-3p (miRNA-329-3p), and the SMAD2 gene and their regulatory relationships in human cervical cancer tissue and cervical cancer cell lines. MATERIAL AND METHODS Cervical cancer tissue samples (n=30) and normal control cervical tissues were studied. Cell proliferation and migration were investigated in HeLa and SiHa human cervical cancer cells using the MTT assay, crystal violet staining, wound healing assay, and the transwell assay. Expression of lncRNA TP73-AS1 and the SMAD2 gene were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Enrichment of miR-329-3p was measured using the RNA immunoprecipitation assay (RIPA). Targeting relationships between TP73-AS1, miR-329-3p, and SMAD2 were identified using the dual-luciferase reporter assay. A subcutaneous xenograft model was established, tumor size was measured, and SMAD2 expression was detected using immunohistochemistry. RESULTS LncRNA TP73-AS1 was overexpressed in cervical cancer tissues and cells and was associated with reduced expression of miR-329-3p. Down-regulation of lncRNA TP73-AS1 inhibited cell proliferation, migration and invasion and increased miR-329-3p expression. Expression of SMAD2 down-regulated miR-329-3p and was associated with increased expression of TP73-AS1. LncRNA TP73-AS1 knockdown resulted in miR-329-3p silencing. In tumor xenografts, expression of TP73-AS1 reduced the tumor volume and down-regulated the expression levels of the SMAD2 gene. CONCLUSIONS LncRNA TP73-AS1 promoted proliferation of cervical cancer cell lines by targeting miR-329-3p to regulate the expression of the SMAD2 gene. A regulatory network was formed between lncRNA TP73-AS1, miR-329-3p, and SMAD2.


Long-term care liability for pressure ulcers.
More than 20% of residents who have been in long-term care (LTC) facilities for 2 or more years will develop at least one pressure ulcer (PU). Residents suffer pain, disfigurement, and decreased quality of life, and their risk of illness and death increases. LTC facilities face censure from residents, their families, and surveyors and the threat of expensive lawsuits. Lawsuits are typically based on contentions of residents with a PU--or their advocates--that the LTC facility was negligent and failed to provide the care that, by industry standards, it must provide to prevent or manage such wounds (managing pressure, incontinence, and nutrition). In this article, data from 1999 and 2002 are presented, showing that lawsuits related to PUs are increasingly common and costly for LTC owners and care providers. Residents realized some type of recovery against the facility in 87% of the cases (verdicts for the resident plus settlements) and were awarded amounts as high as $312 million in damages. Even LTC administrators who believe that care in their facility equals or exceeds industry standards often settle lawsuits out of court to avoid jury verdicts. The data also show that jury awards were highest for PUs caused by multiple factors and that the highest awards for PUs caused by a single factor were seen when that factor was inadequate nutrition. LTC providers can help improve the health and quality of life of their residents, improve survey results, and minimize their risk of expensive lawsuits by developing, implementing, and documenting a plan of basic measures to prevent PUs.


Helicobacter pylori-related disease: guidelines for testing and treatment.
To develop practical guidelines for the treatment of patients with suspected and documented Helicobacter pylori-related gastroduodenal diseases. A panel of physicians with expertise in H. pylori reviewed, critically appraised, and synthesized the literature on assigned topics and presented their overviews to the panel. Consensus was obtained in controversial areas through discussion. The panel recommended testing for H. pylori in patients with active ulcers, a history of ulcers, or gastric mucosa-associated lymphoid tissue lymphomas. Young, otherwise healthy patients with ulcerlike dyspepsia and those with a family history or fear of gastric cancer may also undergo H pylori testing. Non-endoscopic methods are preferred for H. pylori diagnosis. Dual medication regimens should not be used for therapy; twice-daily triple therapy with a proton pump inhibitor or ranitidine bismuth citrate, clarithromycin, and amoxicillin for 10 to 14 days is an appropriate therapy. Posttreatment assessment of H. pylori status using urea breath testing should be considered in patients with a documented history of ulcer disease or with persistent symptoms.


Antioxidant effects of extracts from Cassia tora L. prepared under different degrees of roasting on the oxidative damage to biomolecules.
The effects of water extracts from Cassia tora L. (WECT) treated with different degrees of roasting (unroasted and roasted at 150, 200, and 250 degrees C) on the oxidative damage to deoxyribose, DNA, and DNA base in vitro were investigated. It was found that WECT alone induced a slight strand breaking of DNA. In the presence of Fe(3+)/H(2)O(2), WECT accelerated the strand breaking of DNA at a concentration of 2 microg/mL; however, it decreased with increasing concentrations (>5 microg/mL) of WECT. WECT also accelerated the oxidation of deoxyribose induced by Fe(3+)-EDTA/H(2)O(2) at a concentration of 0.2 mg/mL but inhibited the oxidation of deoxyribose induced by Fe(3+)-EDTA/H(2)O(2)/ascorbic acid. Furthermore, WECT accelerated the oxidation of 2'-deoxyguanosine (2'-dG) to form 8-OH-2'-dG induced by Fe(3+)-EDTA/H(2)O(2). The prooxidant action of WECT on the oxidation of 2'-dG was in the order of unroasted > roasted at 150 degrees C > roasted at 200 degrees C > roasted at 250 degrees C. The decrease in the prooxidant activity of the roasted sample might be due to the reduction in its anthraquinone glycoside content or the formation of antioxidant Maillard reaction products after roasting. Thus, WECT exhibited either a prooxidant or an antioxidant property in the model system that was dependent on the activities of the reducing metal ions, scavenging hydroxyl radical, and chelating ferrous ion.


Trends in adolescent emotional problems in Wales between 2013 and 2019: the contribution of peer relationships.
Epidemiological evidence shows a substantial increase in adolescent emotional problems in many countries, but reasons for this increase remain poorly understood. We tested change in emotional problems in a national sample of young people in Wales in 2013, 2017 and 2019 using identical symptom screens, and examined whether trends were accounted for by changes in youth friendship quality and bullying. The present study of 230,735 11-16-year olds draws on repeat cross-sectional data obtained on three occasions (2013, 2017 and 2019) in national school-based surveys in Wales (conducted by the School Health Research Network). Emotional problems were assessed with a brief validated symptom screen (the SCL-4). There was a significant increase in emotional problem scores between 2013 and 2019 (b[95% CI] = 1.573 [1.380, 1.765]). This increase was observed for all ages and was more pronounced for girls than boys (interaction b [95% CI] = 0.229 [0.004, 0.462]) and for young people from less affluent families (interaction b [95% CI] = -0.564[-0.809, -0.319]). Of the total sample, 14.2% and 5.7% reported frequent face-to-face and cyberbullying respectively. There were modest decreases in friendship quality and increases in rates of bullying between 2013 and 2019, but accounting for these changes did not attenuate estimates of the population-level increase in emotional problems. This study provides evidence of a substantial increase in emotional problems among young people in Wales, particularly for young people from less affluent backgrounds. Changes in bullying or friendship quality did not explain this increase.


Temperament and character traits in major depressive disorder: a case control study.
Patients with major depressive disorder (MDD) have distinct personality traits, compared with control subjects, although the role of anxiety and positive and negative affects in this finding is unclear. A case-control study enrolling 103 antidepressant-free depressed patients and 103 age and gender-matched controls was conducted at the University Hospital, University of São Paulo. The self-reported scales of the Positive and Negative Affect Schedule (PANAS), State-Trait Anxiety Inventory (STAI) and Cloninger's Temperament and Character Inventory (TCI) were applied. Temperament and character traits were compared between groups using multivariate and bivariate analyses of variance (MANOVA and ANOVA). The influence of anxiety and affect was further investigated using ANOVA and mediation analyses. Depressed patients presented higher harm avoidance and lower self-directedness scores than controls. After adjustment for anxiety trait, harm avoidance was no longer significantly different between groups. Mediation analysis revealed that the anxiety trait, but not state-anxiety or affect, fully mediated the influence of group (depressed versus control subjects) on harm avoidance. Our findings confirm that depressed patients present personality traits distinct from those of controls and suggest that MDD is not directly associated with harm avoidance, but that this effect is fully mediated through the anxiety trait.


"It's Embedded in What We Do for Every Child": A Qualitative Exploration of Early Childhood Educators' Perspectives on Supporting Children's Social and Emotional Learning.
Early childhood educators play an important role in supporting children's social and emotional development. While a growing body of research has examined the impact of curriculum-based social and emotional learning (SEL) programs on child outcomes, the approaches educators use to strengthen children's social and emotional functioning through their everyday practices are less defined. This study explored Australian early childhood educators' perspectives on children's social and emotional development, the approaches educators use to encourage children's social and emotional skills, the enablers and barriers to SEL within the preschool environment, and the additional support needed. Thirty Early Childhood Education and Care professionals participated in semi-structured interviews and focus group discussions. Findings suggest children's social-emotional development is at the forefront of educator planning, practice, and reflection. Participants described utilising various approaches to support children's social and emotional skills, embedded within interactions and relationships with children and families. Specifically, strategies could be grouped into four broad categories: a nurturing and responsive educator-child relationship; supporting SEL through everyday interactions and practice; utilising the physical environment to encourage SEL; and working in partnership with caregivers. There was, however, inconsistency in the variety and type of approaches identified. Time constraints, group size, educator confidence and capability, high staff turnover, and limited guidance regarding high-quality social and emotional pedagogy were identified as key barriers. Participants sought practical strategies that could be embedded into daily practice to build upon current knowledge.


Ecological Influences and Morphological Correlates of Resting and Maximal Metabolic Rates across Teleost Fish Species.
Rates of aerobic metabolism vary considerably across evolutionary lineages, but little is known about the proximate and ultimate factors that generate and maintain this variability. Using data for 131 teleost fish species, we performed a large-scale phylogenetic comparative analysis of how interspecific variation in resting metabolic rates (RMRs) and maximum metabolic rates (MMRs) is related to several ecological and morphological variables. Mass- and temperature-adjusted RMR and MMR are highly correlated along a continuum spanning a 30- to 40-fold range. Phylogenetic generalized least squares models suggest that RMR and MMR are higher in pelagic species and that species with higher trophic levels exhibit elevated MMR. This variation is mirrored at various levels of structural organization: gill surface area, muscle protein content, and caudal fin aspect ratio (a proxy for activity) are positively related with aerobic capacity. Muscle protein content and caudal fin aspect ratio are also positively correlated with RMR. Hypoxia-tolerant lineages fall at the lower end of the metabolic continuum. Different ecological lifestyles are associated with contrasting levels of aerobic capacity, possibly reflecting the interplay between selection for increased locomotor performance on one hand and tolerance to low resource availability, particularly oxygen, on the other. These results support the aerobic capacity model of the evolution of endothermy, suggesting elevated body temperatures evolved as correlated responses to selection for high activity levels.


Orbital malignant schwannoma and embryonal rhabdomyosarcoma in rats caused by leakage of locally injected nickel subsulfide to the orbit.
Nickel subsulfide (Ni3S2) is known to induce intraocular neoplasms when injected intravitreally into the eyes of rats. Here, we found two extraocular orbital neoplasms in two different rat strains, presumably due to the leakage of locally injected Ni3S2 to the extraocular orbital tissues. In the F344/DuCrlCrlj rat, an orbital mass arose at 30 weeks after injection, and invaded into the cranium. Histologically, the orbital mass was composed of areas arranged in parallel bundles formed by densely packed elongated or spindle-shaped cells with indistinct cytoplasmic borders, and of areas of hypocellular arrangement consisting of round cells in eosinophilic myxoid-like substances. Metastases were observed in the right submandibular and cervical lymph nodes. The neoplastic cells were immunopositive for S-100 protein and vimentin. Transmission electron microscopy revealed that the neoplastic cells had cellular processes and pericytoplasmic basal laminae. In the RccHanTM:WIST rat, an orbital mass arose at 36 weeks after injection. Histologically, the mass consisted of rhabdoid-like large round cells with proliferation of small round-to-polygonal cells, and these neoplastic cells infiltrated into the extraocular muscles. Immunohistochemically, the neoplastic cells were positive for desmin and vimentin. Transmission electron microscopy detected immature myofibrils with Z-band structures in the cytoplasm of these neoplastic cells. Consequently, the tumors were diagnosed as an orbital malignant schwannoma in an F344/DuCrlCrlj rat and an orbital embryonal rhabdomyosarcoma in a RccHanTM:WIST rat. The results of this case report suggest that leakage of Ni3S2 to the orbit caused the induction of orbital malignant schwannoma or rhabdomyosarcoma in rats.


Supplementation with selenium and coenzyme Q10 in critically ill patients.
Multiple organ dysfunction and resultant mortality in critically ill patients has been linked with impaired cellular energy supply and oxidative stress. Clinical studies supplementing selenium, on the basis of its role as a key cofactor of antioxidant enzymes, have reported variable outcomes in critically ill patients. However, the synergistic interaction between selenium and coenzyme Q10, which has essential roles in cellular energy supply and as an antioxidant, has not been considered in such studies. This article reviews the link between selenium and coenzyme Q10, and the potential role of their co-supplementation in critical illness.


Detection of K-ras mutations in pancreatic and hepatic neoplasms by non-isotopic mismatched polymerase chain reaction.
Mutations within codon 12 leading to activation of Kirsten-ras (K-ras) genes occur in a wide variety of human tumors, but have been reported most frequently in pancreatic carcinomas. We studied twenty-four paraffin-embedded pancreatic and hepatic tumors and two colon carcinoma cell lines with a rapid and simple approach that exploits allele-specific amplification of genomic DNA in a polymerase chain reaction (PCR). We extend the utility of this technique, which is dependent on an exact match at the 3' nucleotide between synthetic oligonucleotides and template DNA, to analyse paraffin-embedded tumor samples for the presence of point mutations at the first and second base of codon 12 of the K-ras gene. The PCR mismatch amplification technique demonstrated a 66% incidence of K-ras mutations at codon 12 in the group of pancreatic neoplasms as a whole. The percentage of mutations varied only slightly in the pancreatic cancer subcategories: 75% in ampullary, 66% in bile duct and 57% in the ductal adenocarcinomas. One islet cell carcinoma and normal tissues adjacent to the tumors revealed wild-type alleles only. One hepatoblastoma and one of six hepatocellular carcinomas also had codon 12 mutations. The PCR mismatch is a sensitive and rapid method that may be useful in screening neoplasms for K-ras point mutation and can be applied to archival material. This application allows a retrospective analyses of a wide range of pathological specimens to determine the role of K-ras mutations in human tumorigenesis.


DNA methylation is a primary mechanism for silencing postmigratory primordial germ cell genes in both germ cell and somatic cell lineages.
DNA methylation is necessary for the silencing of endogenous retrotransposons and the maintenance of monoallelic gene expression at imprinted loci and on the X chromosome. Dynamic changes in DNA methylation occur during the initial stages of primordial germ cell development; however, all consequences of this epigenetic reprogramming are not understood. DNA demethylation in postmigratory primordial germ cells coincides with erasure of genomic imprints and reactivation of the inactive X chromosome, as well as ongoing germ cell differentiation events. To investigate a possible role for DNA methylation changes in germ cell differentiation, we have studied several marker genes that initiate expression at this time. Here, we show that the postmigratory germ cell-specific genes Mvh, Dazl and Scp3 are demethylated in germ cells, but not in somatic cells. Premature loss of genomic methylation in Dnmt1 mutant embryos leads to early expression of these genes as well as GCNA1, a widely used germ cell marker. In addition, GCNA1 is ectopically expressed by somatic cells in Dnmt1 mutants. These results provide in vivo evidence that postmigratory germ cell-specific genes are silenced by DNA methylation in both premigratory germ cells and somatic cells. This is the first example of ectopic gene activation in Dnmt1 mutant mice and suggests that dynamic changes in DNA methylation regulate tissue-specific gene expression of a set of primordial germ cell-specific genes.


A phase II study of vindesine in the treatment of esophageal carcinoma. Japanese Esophageal Oncology Group.
A phase II study of Vindesine for esophageal carcinoma was carried out cooperatively by 10 Japanese institutions. Fifty patients were enrolled in the study over a year and ten months. Four patients could not be evaluated because less than 4 weeks had elapsed since their preceding therapy. There were one complete remission and five partial remissions among the 46 remaining patients, a response rate of 13.0%. The patient experiencing the complete remission is still alive more than 24 months after treatment. The main side effects were depilation, anorexia, stomatitis and leukopenia. Thrombocytopenia and elevations of blood urea nitrogen and creatinine were not marked. There was no mortality connected with the administration of Vindesine. It was concluded that Vindesine is useful in the treatment of esophageal carcinoma as a single agent.


Second-Day Morning Cortisol Levels after Transsphenoidal Surgery Are Accurate Predictors of Secondary Adrenal Insufficiency with Diagnostic Cut-Offs Similar to Those in Non-Stressed Conditions.
Multiple studies tried to identify cortisol cut-offs after pituitary surgery that could accurately assess hypothalamic-pituitary-adrenal (HPA) axis function; however, there is no consensus nowadays. This study aimed to evaluate the accuracy of morning cortisol after transsphenoidal surgery in predicting long-term secondary adrenal insufficiency. In our tertiary center, we prospectively determined first- and second-day cortisol after transsphenoidal surgery in 92 patients without preoperative adrenal -insufficiency and not treated with glucocorticoids perioperatively. Definitive diagnosis of secondary adrenal insufficiency was obtained with re-evaluation 3 months after trans-sphenoidal surgery and clinical follow-up of at least 1 year. Ten patients (10.8%) developed long-term postoperative secondary adrenal insufficiency. The ROC curves demonstrated that first-day cortisol had a moderate diagnostic accuracy, while a second-day cortisol ≤9.3 µg/dL (257 nmol/L) showed the best performance in predicting adrenal insufficiency (sensitivity [Se] 88.9%, specificity [Sp] 86.9%, AUC 0.921). Moreover, a second-day cortisol ≤3.2 µg/dL (89 nmol/L) was able to diagnose adrenal insufficiency in 100% of cases (Se 22.2%, Sp 100%) and >14 µg/dL (386 nmol/L) was able to exclude ACTH deficiency (Se 100%, Sp 57.4%). Adrenal function can be carefully studied on the second day after pituitary surgery, using cut-off values that international guidelines suggested for non-stressed conditions. In fact, second-day cortisol levels ≤3.2 μg/dL (89 nmol/L) and >14 μg/dL (386 nmol/L) are diagnostic of secondary adrenal insufficiency and normal function, respectively. We also suggest performing a definitive re-evaluation with an HPA axis stimulation test when second-day cortisol values are between 3.3 and 14 μg/dL (90-386 nmol/L).


A quasi-integral controller for adaptation of genetic modules to variable ribosome demand.
The behavior of genetic circuits is often poorly predictable. A gene's expression level is not only determined by the intended regulators, but also affected by changes in ribosome availability imparted by expression of other genes. Here we design a quasi-integral biomolecular feedback controller that enables the expression level of any gene of interest (GOI) to adapt to changes in available ribosomes. The feedback is implemented through a synthetic small RNA (sRNA) that silences the GOI's mRNA, and uses orthogonal extracytoplasmic function (ECF) sigma factor to sense the GOI's translation and to actuate sRNA transcription. Without the controller, the expression level of the GOI is reduced by 50% when a resource competitor is activated. With the controller, by contrast, gene expression level is practically unaffected by the competitor. This feedback controller allows adaptation of genetic modules to variable ribosome demand and thus aids modular construction of complicated circuits.


Involvement of ERK and p38 MAPK pathways on Interleukin-33-induced RANKL expression in osteoblastic cells.
The receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG) system is a well-known key factor in osteoclast differentiation, and osteoblastic lineage cells are the major sources of RANKL and OPG in local bone tissue. Recently, a new molecule from the interleukin (IL)-1 family, IL-33, was identified. Here, we report the possible involvement of IL-33 in RANKL and OPG expression, and the signaling pathways that are required for maximal IL-33-induced RANKL expression in MC3T3-E1 osteoblastic cells. Stimulation with IL-33 increased the mRNA expression and secretion of RANKL in MC3T3-E1 cells. The IL-33-induced RANKL mRNA expression was inhibited by an anti-IL-33 monoclonal antibody. Furthermore, ERK and p38 MAPK inhibitors, but not a JNK inhibitor, suppressed IL-33-induced RANKL mRNA expression. On the other hand, IL-33 had no effect on OPG mRNA expression and protein secretion. These results taken together suggest that IL-33 stimulates RANKL expression through mechanisms dependent on the ERK and p38 MAPK pathways in MC3T3-E1 cells.


How to manage traumatic brain injury without invasive monitoring?
Severe traumatic brain injury (TBI) is an extremely serious health problem, especially in low-middle income countries (LMICs). The prevalence of severe TBI continues to increase in LMICs. Major limitations in the chain of care for TBI patients are common in LMICs including suboptimal or nonexistent prehospital care, overburdened emergency services, lack of trained human resources and limited availability of ICUs. Basic neuromonitoring, such as intracranial pressure, are unavailable or underutilized and advanced techniques are not available. Attention to fundamental principles of TBI care in LMICs, including early categorization, prevention and treatment of secondary insults, use of low-cost technology for evaluation of intracranial bleeding and neuromonitoring, and emphasis on education of human resources and multidisciplinary work, are particularly important in LMICs. Institutional collaborations between high-income and LMICs have developed evidence focused on available resources. Accordingly, an expert group have proposed consensus recommendations for centers without availability of invasive brain monitoring. Severe TBI is very prevalent in LMIC and neuromonitoring is often not available in these environments. When intracranial pressure monitors are not available, careful attention to changes on clinical examination, serial imaging and noninvasive monitoring techniques can help recognize intracranial hypertension and effectively guide treatment decisions.


[Development of antisocial disorders--impact of the autonomic stress system].
The current selective review emphasizes the heterogeneity of antisocial behaviour in children and adolescents. It focuses on the development of children of the early-starter subtype of conduct disorder who are at high risk for the development of an antisocial personality disorder. Especially the autonomic stress system seems to have an important impact on symptoms and the prognosis of antisocial individuals. While autonomic hypoarousal and a reduced autonomic reagibility seem to be associated with more proactive aggressiveness and a negative outcome, increased autonomic arousal and reagibility might be related to reactive aggressiveness and constitutes possibly a protective trait. Data of the current psychophysiological and neuroendocrinological literature are summarized. Moreover, the impact of comorbid attention deficit hyperactivity disorder and anxiety disorders on dissocial development is illustrated. Particularly early diagnostic assessment of the individual's extent of trait anxiety might help to specify therapeutic opportunities and could thereby improve therapeutic efficiency.


Activity spaces in studies of the environment and physical activity: A review and synthesis of implications for causality.
Activity spaces are increasingly used to understand how people interact with their environment and engage in activity but their use may raise challenges regarding causal inference. We conducted a systematic review of findings and the methodological, analytical and conceptual issues relevant to causal inference. Studies were included if they comprised a spatial summary of locations visited, assessed any part of the causal pathway between the environment, physical activity and health, and used quantitative or qualitative methods. We searched seven electronic databases in January 2018 and screened 11910 articles for eligibility. Forty-seven studies were included for review. Studies answered research questions about features of or environmental features within activity spaces using a range of spatial and temporal summary techniques. The conceptual challenge of using activity spaces to strengthen causal inference was rarely considered, although some studies discussed circularity, temporality, and plausibility. Future studies should use longitudinal and experimental designs and consider the potential and actual use of spaces for physical activity, and their relationship with total levels of activity.


Machine Learning-Enabled NIR Spectroscopy. Part 3: Hyperparameter by Design (HyD) Based ANN-MLP Optimization, Model Generalizability, and Model Transferability.
Data variations, library changes, and poorly tuned hyperparameters can cause failures in data-driven modelling. In such scenarios, model drift, a gradual shift in model performance, can lead to inaccurate predictions. Monitoring and mitigating drift are vital to maintain model effectiveness. USFDA and ICH regulate pharmaceutical variation with scientific risk-based approaches. In this study, the hyperparameter optimization for the Artificial Neural Network Multilayer Perceptron (ANN-MLP) was investigated using open-source data. The design of experiments (DoE) approach in combination with target drift prediction and statistical process control (SPC) was employed to achieve this objective. First, pre-screening and optimization DoEs were conducted on lab-scale data, serving as internal validation data, to identify the design space and control space. The regression performance metrics were carefully monitored to ensure the right set of hyperparameters was selected, optimizing the modelling time and storage requirements. Before extending the analysis to external validation data, a drift analysis on the target variable was performed. This aimed to determine if the external data fell within the studied range or required retraining of the model. Although a drift was observed, the external data remained well within the range of the internal validation data. Subsequently, trend analysis and process monitoring for the mean absolute error of the active content were conducted. The combined use of DoE, drift analysis, and SPC enabled trend analysis, ensuring that both current and external validation data met acceptance criteria. Out-of-specification and process control limits were determined, providing valuable insights into the model's performance and overall reliability. This comprehensive approach allowed for robust hyperparameter optimization and effective management of model lifecycle, crucial in achieving accurate and dependable predictions in various real-world applications.


Atypical teratoid/rhabdoid tumor of the central nervous system: a comparative study with primitive neuroectodermal tumor/medulloblastoma.
Eleven atypical teratoid/rhabdoid tumors (AT/RT) and 121 primitive neuroectodermal tumors/medulloblastomas (PNET/MB) were included in this study for evaluation of the histopathological features of AT/RT and comparison between AT/RT and PNET/MB. Histopathological studies of AT/RT showed that in addition to the commonly recognized components, i.e., rhabdoid cells, small (PNET/MB) cells, spindle cells and epithelial components, there was a previously unrecognized component, sickle-shaped embracing cells, which were present in all cases and could be useful as a histological marker of this tumor. Immunohistochemical studies showed divergent differentiation of the tumor cells and among the 16 antibodies studied, vimentin, neuron-specific enolase, epithelial membrane antigen and glial fibrillary acidic protein were most commonly reactive. The frequency of AT/RT expressed as a ratio of AT/RT to PNET/MB was 1:11 in general and increased to 1:3.8 among patients younger than 3 years old. The AT/RT patients were younger than those with PNET/MB and had a female predominance. The MIB-1 labeling index of AT/RT was significantly higher than that of PNET/MB (mean 63.9 vs 40.1), which correlated with a shorter survival in patients with AT/RT than those with PNET/MB (median survival time 15.4 months vs 156.4 months).


Controlling the outcome of an N-alkylation reaction by using N-oxide functional groups.
Covalent modifiers of proteins are of importance in chemical proteomics, an emerging chemical technology used to assign protein function. In this study, high-field (1)H NMR techniques were used to analyze the reaction of the bioactive compound, 2,3-bis(bromomethyl)quinoxaline 1,4-dioxide, with amines (a model system for proteins containing nitrogen-based nucleophiles). Unexpectedly, the results show that a double nucleophilic substitution reaction involving 2 equiv of the amine is preferred to an intramolecular cyclization pathway. A direct comparison with the reaction carried out on a substrate lacking the N-oxide functional groups is also provided. X-ray crystal structures and computational studies are used to rationalize the observed differences in reactivity between the two systems.


An SH2 domain-dependent, phosphotyrosine-independent interaction between Vav1 and the Mer receptor tyrosine kinase: a mechanism for localizing guanine nucleotide-exchange factor action.
Mer belongs to the Mer/Axl/Tyro3 receptor tyrosine kinase family, which regulates immune homeostasis in part by triggering monocyte ingestion of apoptotic cells. Mutations in Mer can also cause retinitis pigmentosa, again due to defective phagocytosis of apoptotic material. Although, some functional aspects of Mer have been deciphered, how receptor activation lead to the physiological consequences is not understood. By using yeast two-hybrid assays, we identified the carboxyl-terminal region of the guanine nucleotide-exchange factor (GEF) Vav1 as a Mer-binding partner. Unlike similar (related) receptors, Mer interacted with Vav1 constitutively and independently of phosphotyrosine, yet the site of binding localized to the Vav1 SH2 domain. Mer activation resulted in tyrosine phosphorylation of Vav1 and release from Mer, whereas Vav1 was neither phosphorylated nor released from kinase-dead Mer. Mutation of the Vav1 SH2 domain phosphotyrosine coordinating Arg-696 did not alter Mer/Vav1 constitutive binding or Vav1 tyrosine phosphorylation but did retard Vav1 release from autophosphorylated Mer. Ligand-dependent activation of Mer in human monocytes led to Vav1 release and stimulated GDP replacement by GTP on RhoA family members. This unusual constitutive, SH2 domain-dependent, but phosphotyrosine-independent, interaction and its regulated local release and subsequent activation of Rac1, Cdc42, and RhoA may explain how Mer coordinates precise cytoskeletal changes governing the ingestion of apoptotic material by macrophages and pigmented retinal epithelial cells.


Dose-response association between physical activity and sedentary time categories on ageing biomarkers.
Physical activity and sedentary behaviour have been suggested to independently affect a number of health outcomes. To what extent different combinations of physical activity and sedentary behaviour may influence physical function and frailty outcomes in older adults is unknown. The aim of this study was to examine the combination of mutually exclusive categories of accelerometer-measured physical activity and sedentary time on physical function and frailty in older adults. 771 older adults (54% women; 76.8 ± 4.9 years) from the Toledo Study for Healthy Aging participated in this cross-sectional study. Physical activity and sedentary time were measured by accelerometry. Physically active was defined as meeting current aerobic guidelines for older adults proposed by the World Health Organization. Low sedentary was defined as residing in the lowest quartile of the light physical activity-to-sedentary time ratio. Participants were then classified into one of four mutually exclusive movement patterns: (1) 'physically active & low sedentary', (2) 'physically active & high sedentary', (3) 'physically inactive & low sedentary', and (4) 'physically inactive & high sedentary'. The Short Physical Performance Battery was used to measure physical function and frailty was assessed using the Frailty Trait Scale. 'Physically active & low sedentary' and 'physically active & high sedentary' individuals had significantly higher levels of physical function (β = 1.73 and β = 1.30 respectively; all p < 0.001) and lower frailty (β = - 13.96 and β = - 8.71 respectively; all p < 0.001) compared to 'physically inactive & high sedentary' participants. Likewise, 'physically inactive & low sedentary' group had significantly lower frailty (β = - 2.50; p = 0.05), but significance was not reached for physical function. We found a dose-response association of the different movement patterns analysed in this study with physical function and frailty. Meeting the physical activity guidelines was associated with the most beneficial physical function and frailty profiles in our sample. Among inactive people, more light intensity relative to sedentary time was associated with better frailty status. These results point out to the possibility of stepwise interventions (i.e. targeting less strenuous activities) to promote successful aging, particularly in inactive older adults.


N-methylpurine DNA glycosylase and 8-oxoguanine dna glycosylase metabolize the antiviral nucleoside 2-bromo-5,6-dichloro-1-(beta-D-ribofuranosyl)benzimidazole.
The rapid in vivo degradation of the potent human cytomegalovirus inhibitor 2-bromo-5,6-dichloro-1-(beta-D-ribofuranosyl)benzimidazole (BDCRB) compared with a structural L-analog, maribavir (5,6-dichloro-2-(isopropylamino)-1-beta-L-ribofuranosyl-1H-benzimidazole), has been attributed to selective glycosidic bond cleavage. An enzyme responsible for this selective BDCRB degradation, however, has not been identified. Here, we report the identification of two enzymes, 8-oxoguanine DNA glycosylase (OGG1) and N-methylpurine DNA glycosylase (MPG), that catalyze N-glycosidic bond cleavage of BDCRB and its 2-chloro homolog, 2,5,6-trichloro-1-(beta-D-ribofuranosyl)benzimidazole, but not maribavir. To our knowledge, this is the first demonstration that free nucleosides are substrates of OGG1 and MPG. To understand how these enzymes might process BDCRB, docking and molecular dynamics simulations were performed with the native human OGG1 crystal coordinates. These studies showed that OGG1 was not able to bind a negative control, guanosine, yet BDCRB and maribavir were stabilized through interactions with various binding site residues, including Phe319, His270, Ser320, and Asn149. Only BDCRB, however, achieved orientations whereby its anomeric carbon, C1', could undergo nucleophilic attack by the putative catalytic residue, Lys249. Thus, in silico observations were in perfect agreement with experimental observations. These findings implicate DNA glycosylases in drug metabolism.


Tips, Tricks, and Technique for Laparoscopic Colectomy.
In the United States, there are in excess of 300,000 operations for diseases of the colon yearly. Minimally invasive colectomy became a reality early in the 21st century with the advent of laparoscopic colectomy. The goal of minimally invasive colectomy is to improve postoperative pain control, decrease length of hospital stay, decrease recovery time, decrease complications, and thereby decrease the cost of colon resections. There are many facets to laparoscopic colectomy, including completely laparoscopic approach versus hand-assisted approach and the medial versus lateral approach. These decisions are often based on the disease process, surgeon preference and comfort with technique, and patient considerations such as weight and prior operations. This article outlines the pros and cons of each of these factors.


[Evaluation of survival of the pedicle flap by polarographic method].
The data obtained by the authors allowed the use of a polarographic study of pO2 in the pedicled graft tissue as an adjunct method of determining the graft patency. This method may be especially valuable in cases of a complicated period after suturing the graft to a defect, when considerable difficulties arise as to finding the optimum terms for a safe incision of the feeding pedicle.


Formation of supported lipid bilayers by vesicle fusion: effect of deposition temperature.
We have investigated the effect of deposition temperature on supported lipid bilayer formation via vesicle fusion. By using several complementary surface-sensitive techniques, we demonstrate that despite contradicting literature on the subject, high-quality bilayers can be formed below the main phase-transition temperature of the lipid. We have carefully studied the formation mechanism of supported DPPC bilayers below and above the lipid melting temperature (Tm) by quartz crystal microbalance and atomic force microscopy under continuous flow conditions. We also measured the structure of lipid bilayers formed below or above Tm by neutron reflection and investigated the effect of subsequent cooling to below the Tm. Our results clearly show that a continuous supported bilayer can be formed with high surface coverage below the lipid Tm. We also demonstrate that the high dissipation responses observed during the deposition process by QCM-D correspond to vesicles absorbed on top of a continuous bilayer and not to a surface-supported vesicular layer as previously reported.


Plasma oxytocin concentrations following MDMA or intranasal oxytocin in humans.
MDMA (±3,4-methylenedioxymethamphetamine, 'ecstasy') is reportedly used recreationally because it increases feelings of sociability and interpersonal closeness. Prior work suggests that the pro-social effects of MDMA may be mediated by release of oxytocin. A direct examination of plasma levels of oxytocin after acute doses of oxytocin and MDMA, in the same individuals, would provide further evidence for the idea that MDMA produces its pro-social effects by increasing oxytocin. Fourteen healthy MDMA users participated in a 4-session, double-blind study in which they received oral MDMA (0.75 and 1.5mg/kg), intranasal oxytocin (20IU or 40IU), and placebo. Plasma oxytocin concentrations, as well as cardiovascular and subjective effects were assessed before and at several time points after drug administration. MDMA (1.5mg/kg only) increased plasma oxytocin levels to a mean peak of 83.7pg/ml at approximately 90-120min, compared to 18.6pg/ml after placebo. Intranasal oxytocin (40IU, but not 20IU) increased plasma oxytocin levels to 48.0pg/ml, 30-60min after nasal spray administration. MDMA dose-dependently increased heart rate, blood pressure, feelings of euphoria (e.g., 'High' and 'Like Drug'), and feelings of sociability, whereas oxytocin had no cardiovascular or subjective effects. The subjective and cardiovascular responses to MDMA were not related to plasma oxytocin levels, although the N was small for this analysis. Future studies examining the effects of oxytocin antagonists on responses to MDMA will help to determine the mechanism by which MDMA produces pro-social effects.


Outpatient follow-up of nonoperative cerebral contusion and traumatic subarachnoid hemorrhage: does repeat head CT alter clinical decision-making?
Many neurosurgeons obtain repeat head CT at the first clinic follow-up visit for nonoperative cerebral contusion and traumatic subarachnoid hemorrhage (tSAH). The authors undertook a single-center, retrospective study to determine whether outpatient CT altered clinical decision-making. The authors evaluated 173 consecutive adult patients admitted to their institution from April 2006 to August 2012 with an admission diagnosis of cerebral contusion or tSAH and at least 1 clinic follow-up visit with CT. Patients with epidural, subdural, aneurysmal subarachnoid, or intraventricular hemorrhage, and those who underwent craniotomy, were excluded. Patient charts were reviewed for new CT findings, new patient symptoms, and changes in treatment plan. Patients were stratified by neurological symptoms into 3 groups: 1) asymptomatic; 2) mild, nonspecific symptoms; and 3) significant symptoms. Mild, nonspecific symptoms included minor headaches, vertigo, fatigue, and mild difficulties with concentration, short-term memory, or sleep; significant symptoms included moderate to severe headaches, nausea, vomiting, focal neurological complaints, impaired consciousness, or new cognitive impairment evident on routine clinical examination. One hundred seventy-three patients met inclusion criteria, with initial clinic follow-up obtained within approximately 6 weeks. Of the 173 patients, 104 (60.1%) were asymptomatic, 68 patients (39.3%) had mild, nonspecific neurological symptoms, and 1 patient (1.0%) had significant neurological symptoms. Of the asymptomatic patients, 3 patients (2.9%) had new CT findings and 1 of these patients (1.0%) underwent a change in treatment plan because of these findings. This change involved an additional clinic appointment and CT to monitor a 12-mm chronic subdural hematoma that ultimately resolved without treatment. Of the patients with mild, nonspecific neurological symptoms, 6 patients (8.8%) had new CT findings and 3 of these patients (4.4%) underwent a change in treatment plan because of these findings; none of these patients required surgical intervention. The single patient with significant neurological symptoms did not have any new CT findings. Repeat outpatient CT of asymptomatic patients after nonoperative cerebral contusion and tSAH is very unlikely to demonstrate significant new pathology. Given the cost and radiation exposure associated with CT, imaging should be reserved for patients with significant symptoms or focal findings on neurological examination.


Trends in the morbidity of pre-school children registered in family medicine and primary pediatrics in Croatia: 1995-2012.
Primary health care for pre-school children in Croatia is provided by the primary pediatricians (PPs) in urban, or family doctors (FDs) in rural areas. The main aim of the study was to determine the trends in morbidity as recorded by FDs, and those recorded by PPs, from 1995-2012. The study was based on the routinely collected, national statistics data from the Croatian Health Service Yearbooks. The results obtained clearly indicate that there are no differences in the morbidity patterns registered by FDs and PPs. The average annual number of diagnoses per child, and the most frequently registered diagnostic groups were almost the same for both services. The number of children under the particular service was the only difference, with a decreasing trend in family medicine and an increasing in pediatric service. The national routine health statistics system does not provide enough information to permit a deeper analysis of the childhood morbidity in two services.


[Imatinib-induced toxic hepatitis: description of two cases and review of the literature].
Imatinib is currently the treatment of choice in chronic myeloid leukemia. The use of this drug is safe, although some cases of imatinib-induced toxic hepatitis have been reported. We present 2 patients treated with this drug who developed acute anicteric hepatitis months after starting treatment. We also review 20 reports of individual cases to characterize imatinib-induced hepatitis. Imatinib-induced hepatitis has a variable latency period, frequently of several months. Half of the patients develop anicteric hepatitis and the clinical course is generally benign. A distinguishing feature of this entity is a transitory increase in transaminase levels in patients diagnosed with hepatitis in the weeks after treatment withdrawal. Resumption of imatinib use provokes hepatitis recurrence, which can be avoided by simultaneous prednisone administration.


Automatic registration and error detection of multiple slices using landmarks.
When analysing the 3D structure of tissue, serial sectioning and staining of the resulting slices is sometimes the preferred option. This leads to severe registration problems. In this paper, a method for automatic registration and error detection of slices using landmark needles has been developed. A cost function takes some parameters from the current state of the problem to be solved as input and gives a quality of the current solution as output. The cost function used in this paper, is based on a model of the slices and the landmark needles. The method has been used to register slices of prostates in order to create 3D computer models. Manual registration of the same prostates has been undertaken and compared with the results from the algorithm. Prostates from sixteen men who underwent radical prostatectomy were formalin fixed with landmark needles, sliced and the slices were computer reconstructed. The cost function takes rotation and translation for each prostate slice, as well as slope and offset for each landmark needle as input. The current quality of fit of the model, using the input parameters given, is returned. The function takes the built-in instability of the model into account. The method uses a standard algorithm to optimize the prostate slice positions. To verify the result, s standard method in statistics was used. The methods were evaluated for 16 prostates. When testing blindly, a physician could not determine whether the registration shown to him were created by the automated method described in this paper, or manually by an expert, except in one out of 16 cases. Visual inspection and analysis of the outlier confirmed that the input data had been deformed. The automatic detection of erroneous slices marked a few slices, including the outlier, as suspicious. The model based registration performs better than traditional simple slice-wise registration. In the case of prostate slice registration, other aspects, such as the physical slicing method used, may be more important to the final result than the selection of registration method to use.


Critical scaling of whole-brain resting-state dynamics.
Scale invariance is a characteristic of neural activity. How this property emerges from neural interactions remains a fundamental question. Here, we studied the relation between scale-invariant brain dynamics and structural connectivity by analyzing human resting-state (rs-) fMRI signals, together with diffusion MRI (dMRI) connectivity and its approximation as an exponentially decaying function of the distance between brain regions. We analyzed the rs-fMRI dynamics using functional connectivity and a recently proposed phenomenological renormalization group (PRG) method that tracks the change of collective activity after successive coarse-graining at different scales. We found that brain dynamics display power-law correlations and power-law scaling as a function of PRG coarse-graining based on functional or structural connectivity. Moreover, we modeled the brain activity using a network of spins interacting through large-scale connectivity and presenting a phase transition between ordered and disordered phases. Within this simple model, we found that the observed scaling features were likely to emerge from critical dynamics and connections exponentially decaying with distance. In conclusion, our study tests the PRG method using large-scale brain activity and theoretical models and suggests that scaling of rs-fMRI activity relates to criticality.


Graft-versus-host disease management.
Graft-versus-host disease (GVHD) remains a major problem of allogeneic hematopoietic-stem cell transplantation (HSCT) and an obstacle for successful outcome. Clinically significant acute GVHD (grade II or higher) developed in 20 to 65 percent of the patients. Death due to this complication accounts for approximately 50 percent of the deaths that are not due to a relapse of the neoplasm. Up to 70 % of patients who survive beyond day 100 develop chronic GVHD and it is the leading cause of nonrelapse mortality more than 2 years after allogeneic HSCT. In addition, chronic GVHD is associated with decreased quality of life, impaired functional status, and ongoing need for immunosuppressive medications. The incidence of chronic GVHD is increasing because of expansion of the donor population beyond HLA-identical siblings, older recipient age, use of peripheral blood cells as the graft source, and infusion of donor lymphocytes for treatment of recurrent malignancy after HSCT. With the current rush in new findings related to GVHD, we see a significant advancement in its management. Given these various new options and challenges, it is important to identify the minimal requirements for diagnosis and treatment of GVHD, as access to the most sophisticated advances may vary depending on local circumstances (Tab. 4, Fig. 1, Ref. 51).


Methods of usability testing in the development of eHealth applications: A scoping review.
The number of eHealth applications has exponentially increased in recent years, with over 325,000 health apps now available on all major app stores. This is in addition to other eHealth applications available on other platforms such as PC software, web sites and even gaming consoles. As with other digital applications, usability is one of the key factors in the successful implementation of eHealth apps. Reviews of the literature on empirical methods of usability testing in eHealth were last published in 2015. In the context of an exponentially increasing rate of App development year on year, an updated review is warranted. To identify, explore, and summarize the current methods used in the usability testing of eHealth applications. A scoping review was conducted on literature available from April 2014 up to October 2017. Four databases were searched. Literature was considered for inclusion if it was (1) focused on an eHealth application (which includes websites, PC software, smartphone and tablet applications), (2) provided information about usability of the application, (3) provided empirical results of the usability testing, (4) a full or short paper (not an abstract) published in English after March 2014. We then extracted data pertaining to the usability evaluation processes described in the selected studies. 133 articles met the inclusion criteria. The methods used for usability testing, in decreasing order of frequency were: questionnaires (n = 105), task completion (n = 57), 'Think-Aloud' (n = 45), interviews (n = 37), heuristic testing (n = 18) and focus groups (n = 13). Majority of the studies used one (n = 45) or two (n = 46) methods of testing. The rest used a combination of three (n = 30) or four (n = 12) methods of testing usability. None of the studies used automated mechanisms to test usability. The System Usability Scale (SUS) was the most frequently used questionnaire (n = 44). The ten most frequent health conditions or diseases where eHealth apps were being evaluated for usability were the following: mental health (n = 12), cancer (n = 10), nutrition (n = 10), child health (n = 9), diabetes (n = 9), telemedicine (n = 8), cardiovascular disease (n = 6), HIV (n = 4), health information systems (n = 4) and smoking (n = 4). Further iterations of the app were reported in a minority of the studies (n = 41). The use of the 'Think-Aloud' (Pearson Chi-squared test: χ2 = 11.15, p < 0.05) and heuristic walkthrough (Pearson Chi-squared test: χ2 = 4.48, p < 0.05) were significantly associated with at least one further iteration of the app being developed. Although there has been an exponential increase in the number of eHealth apps, the number of studies that have been published that report the results of usability testing on these apps has not increased at an equivalent rate. The number of digital health applications that publish their usability evaluation results remains only a small fraction. Questionnaires are the most prevalent method of evaluating usability in eHealth applications, which provide an overall measure of usability but do not pinpoint the problems that need to be addressed. Qualitative methods may be more useful in this regard. The use of multiple evaluation methods has increased. Automated methods such as eye tracking have not gained traction in evaluating health apps. Further research is needed into which methods are best suited for the different types of eHealth applications, according to their target users and the health conditions being addressed.


Toxoplasma gondii protease inhibitor-1 (TgPI-1) is a novel vaccine candidate against toxoplasmosis.
The Toxoplasma gondii serin protease inhibitor-1 (TgPI-1) is a dense granule antigen that showed to specifically inhibit trypsin, chymotrypsin and neutrophil elastase, suggesting a possible modulatory role during the parasite invasion process and on the development of the innate immune response. To study the immune-protective value of TgPI-1, C3H/HeN mice were immunized with a recombinant form of the antigen rTgPI-1 combined with alum. All immunized mice produced specific anti-rTgPI-1 immunoglobulins, with high IgG antibody titers and a mixed IgG(1)/IgG(2a) response, with predominance of IgG(1) production. The cellular immune response was associated with the production of IFN-gamma and IL-10 cytokines. Vaccinated mice displayed significant protection against an oral challenge either after a lethal infection with Me49 cysts (90% survival vs. 50%) and also after a non-lethal infection (58% reduction in brain parasite load) compared to the non-vaccinated control group. In conclusion, rTgPI-1 elicits a strong specific immune response providing partial protection against both T. gondii acute and chronic infection, so it would be a good candidate in a vaccine against toxoplasmosis, which could be combined with other relevant parasite antigens.


Depression chains in seafloor of contrasting morphology, Atacama Trench margin: a comment on Marsh et al. (2018).
This comment presents acoustic and visual data showing deep seafloor depression chains similar to those reported in Marsh et al. (R. Soc. open sci. 5: 180286), though from a different deep-sea setting. Marsh et al. present data collected during cruise JC120 from polymetallic nodule rich sites within the Clarion-Clipperton Fracture Zone (CCFZ), at water depths of between 3999 and 4258 m. Within this comment, we present data collected with equivalent acoustic and imaging devices on-board the RV Sonne (SO261-March/April 2018) from the Atacama Trench, approximately 4000 m depth, which shows comparable depression chains in the seafloor. In contrast with the CCFZ observations, our study area was wholly free of polymetallic nodules, an observation therefore weakening the 'ballast collection' by deep-sea diving mammals formation hypothesis discussed in their paper. We support their alternate hypothesis that if these features are indeed generated by deep-diving megafauna, then they are more likely the resultant traces of infauna feeding or marks made during opportunistic capture of benthic fish/cephalopods. We observed these potential prey fauna with lander and towed camera systems during the cruise, with example images of these presented here. Both the SO261 and JC120 cruises employed high-resolution sidescan systems at deployment altitudes seldom used routinely until the last few years during scientific deep-sea surveys. Given that both cruises found these depression chains in contrasting physical regions of the East Pacific, they may have a more ubiquitous distribution than at just these sites. Thus, the impacts of cetacean foraging behaviour on deep seafloor communities, and the potential relevance of these prey sources to deep-diving species, should be considered.


Studies on membrane topology, N-glycosylation and functionality of SARS-CoV membrane protein.
The glycosylated membrane protein M of the severe acute respiratory syndrome associated coronavirus (SARS-CoV) is the main structural component of the virion and mediates assembly and budding of viral particles. The membrane topology of SARS-CoV M and the functional significance of its N-glycosylation are not completely understood as is its interaction with the surface glycoprotein S. Using biochemical and immunofluorescence analyses we found that M consists of a short glycosylated N-terminal ectodomain, three transmembrane segments and a long, immunogenic C-terminal endodomain. Although the N-glycosylation site of M seems to be highly conserved between group 1 and 3 coronaviruses, studies using a recombinant SARS-CoV expressing a glycosylation-deficient M revealed that N-glycosylation of M neither influence the shape of the virions nor their infectivity in cell culture. Further functional analysis of truncated M proteins showed that the N-terminal 134 amino acids comprising the three transmembrane domains are sufficient to mediate accumulation of M in the Golgi complex and to enforce recruitment of the viral spike protein S to the sites of virus assembly and budding in the ERGIC.