An evaluation of P0.1 measured in mouth and oesophagus, during carbon dioxide rebreathing in COPD.
The pressure generated 100 ms after the onset of an occluded inspiratory effort (P0.1) is advocated and used as a measure of respiratory centre drive. We have re-examined P0.1, measured simultaneously in the mouth (Pmo0.1) and the oesophagus (Poes0.1), during carbon dioxide rebreathing, in eight patients with severe chronic obstructive pulmonary disease, to see whether either indicates central respiratory drive. Pmo0.1 was identical to Poes0.1 in 4 out of 61, greater than Poes0.1 in 18 out of 61, and less than Poes0.1 in 39 out of 61 measurements (overall Poes0.1-Pmo0.1, median +0.075, range -0.175 to +1.01 kPa). Within a rebreathing run in an individual patient, there was considerable variability in the relationship Pmo0.1/Poes0.1 (0.89 +/- 0.24), coefficient of variation (CoV%) 14.4 +/- 3.7%), in the end-expiratory oesophageal pressure (0.7 +/- 0.54 kPa, CoV% 105 +/- 106%), and in the time delay between the onset of a fall in oesophageal pressure (Poes) from the end-expiratory level to the beginning of inspiration, defined as starting when mouth pressure (Pmo) fell below atmospheric pressure (129 +/- 25 ms, CoV% 22.5 +/- 5.3%). We conclude that the problem of determining the true onset of inspiratory muscle activity from pressure data, and the likelihood that breaths are taken from different lung volumes, make it unlikely that Poes0.1 accurately represents central respiratory drive during rebreathing in chronic obstructive pulmonary disease. Furthermore, Pmo0.1 differed from Poes0.1 during rebreathing, and their relationship was not constant, so that Pmo0.1 is even less likely to be a useful reflection of central nervous system output or respiratory centre drive in such patients.


Do aggressive signals evolve towards higher reliability or lower costs of assessment?
It has been suggested that the evolution of signals must be a wasteful process for the signaller, aimed at the maximization of signal honesty. However, the reliability of communication depends not only on the costs paid by signallers but also on the costs paid by receivers during assessment, and less attention has been given to the interaction between these two types of costs during the evolution of signalling systems. A signaller and receiver may accept some level of signal dishonesty by choosing signals that are cheaper in terms of assessment but that are stabilized with less reliable mechanisms. I studied the potential trade-off between signal reliability and the costs of signal assessment in the corncrake (Crex crex). I found that the birds prefer signals that are less costly regarding assessment rather than more reliable. Despite the fact that the fundamental frequency of calls was a strong predictor of male size, it was ignored by receivers unless they could directly compare signal variants. My data revealed a response advantage of costly signals when comparison between calls differing with fundamental frequencies is fast and straightforward, whereas cheap signalling is preferred in natural conditions. These data might improve our understanding of the influence of receivers on signal design because they support the hypothesis that fully honest signalling systems may be prone to dishonesty based on the effects of receiver costs and be replaced by signals that are cheaper in production and reception but more susceptible to cheating.


Detection of aberrant responding on a personality scale in a military sample: an application of evaluating person fit with two-level logistic regression.
Person-fit assessment is used to identify persons who respond aberrantly to a test or questionnaire. In this study, S. P. Reise's (2000) method for evaluating person fit using 2-level logistic regression was applied to 13 personality scales of the Schedule for Nonadaptive and Adaptive Personality (SNAP; L. Clark, 1996) that had been administered to military recruits (N = 2,026). Results revealed significant person-fit heterogeneity and indicated that for 5 SNAP scales (Disinhibition, Entitlement, Exhibitionism, Negative Temperament, and Workaholism), the scale was more discriminating for some people than for others. Possible causes of aberrant responding were explored with several covariates. On all 5 scales, severe pathology emerged as a key influence on responses, and there was evidence of differential test functioning with respect to gender, ethnicity, or both. Other potential sources of aberrancy were carelessness, haphazard responding, or uncooperativeness. Social desirability was not as influential as expected.


Pharmacology of enalapril in children: a review.
Enalapril is an angiotensin-converting enzyme (ACE) inhibitor that is used for the treatment of (paediatric) hypertension, heart failure and chronic kidney diseases. Because its disposition, efficacy and safety differs across the paediatric continuum, data from adults cannot be automatically extrapolated to children. This review highlights paediatric enalapril pharmacokinetic data and demonstrates that these are inadequate to support with certainty an age-related effect on enalapril/enalaprilat pharmacokinetics. In addition, our review shows that evidence to support effective and safe prescribing of enalapril in children is limited, especially in young children and heart failure patients; studies in these groups are either absent or show conflicting results. We provide explanations for observed differences between age groups and indications, and describe areas for future research.


Gene-environment interaction in tobacco-related cancers.
This review summarizes the carcinogenic effects of tobacco smoke and the basis for interaction between tobacco smoke and genetic factors. Examples of published papers on gene-tobacco interaction and cancer risk are presented. The assessment of gene-environment interaction in tobacco-related cancers has been more complex than originally expected for several reasons, including the multiplicity of genes involved in tobacco metabolism, the numerous substrates metabolized by the relevant genes and the interaction of smoking with other metabolic pathways. Future studies on gene-environment interaction and cancer risk should include biomarkers of smoking dose, along with markers of quantitative historical exposure to tobacco. Epigenetic studies should be added to classic genetic analyses, in order to better understand gene-environmental interaction and individual susceptibility. Other metabolic pathways in competition with tobacco genetic metabolism/repair should be incorporated in epidemiological studies to generate a more complete picture of individual cancer risk associated with environmental exposure to carcinogens.


Safety and efficacy of probiotic lactobacilli in promoting growth in post-weaning Swiss mice.
Lactobacillus rhamnosus MA27/6B and L. acidophilus MA27/6R are strains used in feed as probiotics. Their safety profiles and growth-stimulating properties were investigated via in vivo studies on young Swiss mice. After repeated administrations of different probiotic preparations in drinking water, safety parameters determined from liver, spleen and total weight remained unchanged. The growth-stimulating properties of viable or dead lactobacilli were studied after supplementation in drinking water. The feed intake (FI), water intake and body weight gain (WG) of the animals were compared to those of control mice. The lactobacilli supplementation of a sub-optimal diet made of barley allow recording of measurable growth performance of mice. It significantly increased WG compared to control groups (P < 0.01), by +28.9% and +31.7% for L. rhamnosus MA27/6B and L. acidophilus MA27/6R, respectively. This WG was correlated with a decrease in the consumption index. The effect of the dose ingested was also investigated: 10(8) lactobacilli CFU/mouse/day produced greater WG than 10(2), 10(4) or 10(6) CFU/mouse/day. No significant differences in growth performance parameters were observed between mice fed with 10(8) cells of viable or nonviable preparations. The mouse assay described could be used as a preliminary criterion when screening candidate probiotics for growth performance properties.


Predicting survival outcomes using subsets of significant genes in prognostic marker studies with microarrays.
Genetic markers hold great promise for refining our ability to establish precise prognostic prediction for diseases. The development of comprehensive gene expression microarray technology has allowed the selection of relevant marker genes from a large pool of candidate genes in early-phased, developmental prognostic marker studies. The primary analytical task in such studies is to select a small fraction of relevant genes, typically from a list of significant genes, for further investigation in subsequent studies. We develop a methodology for predicting survival outcomes using subsets of significant genes in prognostic marker studies with microarrays. Key components in this methodology include building prediction models, assessing predictive performance of prediction models, and assessing significance of prediction results. As particular specifications, we assume Cox proportional hazard models with a compound covariate. For assessing predictive accuracy, we propose to use the cross-validated log partial likelihood. To assess significance of prediction results, we apply permutation procedures in cross-validated prediction. As an additional key component peculiar to prognostic prediction, we also consider incorporation of standard prognostic factors. The methodology is evaluated using both simulated and real data. The developed methodology for prognostic prediction using a subset of significant genes can provide new insights based on predictive capability, possibly incorporating standard prognostic factors, in selecting a fraction of relevant genes for subsequent studies.


Change of positive selection pressure on HIV-1 envelope gene inferred by early and recent samples.
HIV-1 infection has been on the rise in Japan recently, and the main transmission route has changed from blood transmission in the 1980s to homo- and/or hetero-sexual transmission in the 2000s. The lack of early viral samples with clinical information made it difficult to investigate the possible virological changes over time. In this study, we sequenced 142 full-length env genes collected from 16 Japanese subjects infected with HIV-1 in the 1980s and in the 2000s. We examined the diversity change in sequences and potential adaptive evolution of the virus to the host population. We used a codon-based likelihood method under the branch-site and clade models to detect positive selection operating on the virus. The clade model was extended to account for different positive selection pressures in different viral populations. The result showed that the selection pressure was weaker in the 2000s than in the 1980s, indicating that it might have become easier for the HIV to infect a new host and to develop into AIDS now than 20 years ago and that the HIV may be becoming more virulent in the Japanese population. The study provides useful information on the surveillance of HIV infection and highlights the utility of the extended clade models in analysis of virus populations which may be under different selection pressures.


Catalytic site interactions in yeast OMP synthase.
The enigmatic kinetics, half-of-the-sites binding, and structural asymmetry of the homodimeric microbial OMP synthases (orotate phosphoribosyltransferase, EC 220.127.116.11) have been proposed to result from an alternating site mechanism in these domain-swapped enzymes [R.W. McClard et al., Biochemistry 45 (2006) 5330-5342]. This behavior was investigated in the yeast enzyme by mutations in the conserved catalytic loop and 5-phosphoribosyl-1-diphosphate (PRPP) binding motif. Although the reaction is mechanistically sequential, the wild-type (WT) enzyme shows parallel lines in double reciprocal initial velocity plots. Replacement of Lys106, the postulated intersubunit communication device, produced intersecting lines in kinetic plots with a 2-fold reduction of kcat. Loop (R105G K109S H111G) and PRPP-binding motif (D131N D132N) mutant proteins, each without detectable enzymatic activity and ablated ability to bind PRPP, complemented to produce a heterodimer with a single fully functional active site showing intersecting initial velocity plots. Equilibrium binding of PRPP and orotidine 5'-monophosphate showed a single class of two binding sites per dimer in WT and K106S enzymes. Evidence here shows that the enzyme does not follow half-of-the-sites cooperativity; that interplay between catalytic sites is not an essential feature of the catalytic mechanism; and that parallel lines in steady-state kinetics probably arise from tight substrate binding.


A case report: Familial glucocorticoid deficiency associated with familial focal segmental glomerulosclerosis.
Familial glucocorticoid deficiency (FGD) is a rare autosomal recessive disorder characterized by isolated glucocorticoid deficiency in the presence of normal plasma renin and aldosterone level. Focal segmental glomerulosclerosis (FSGS) is a form of glomerular disease associated with proteinuria and nephritic syndrome. This is the first case of familial glucocorticoid deficiency associated with familial focal segmental glomerulosclerosis. An eight month old boy presented with increased genital pigmentation. Initial investigation revealed that he was glucocorticoid deficient and was started on hydrocortisone and fludrocortisone with a diagnosis of primary adrenal insufficiency. Later fludrocortisone was withdrawn and he was diagnosed to have isolated glucocorticoid deficiency. He later developed focal segmental glomerulosclerosis for which he underwent renal transplantation at the age of five years. Now at the age of twelve years, this boy is doing well on hydrocortisone treatment. His two siblings and a first degree cousin also had isolated glucocorticoid deficiency. One of the above two siblings died due to renal failure secondary to focal segmental glomerulosclerosis. Patients with familial glucocorticoid deficiency should be carefully followed for development of features of nephrotic syndrome.


Benzyl acetates as attractants for the male oriental fruit fly, Dacus dorsalis, and the male melon fly, Dacus cucurbitae.
Fifty compounds related to benzyl acetate were evaluated quantitatively as attractants to the male oriental fruit fly (Dacus dorsalis) and the male melon fly (Dacus cucurbitae). Thienylmethyl acetate was nearly as attractive as benzyl acetate to both species, but cyclohexylmethyl acetate was completely unattractive, emphasizing the role of the planar aromatic ring in receptor interaction. Although benzyl acetate was equally attractive to both species, para substituents invariably reduced attraction to the oriental fruit fly. A number of derivatives, including p-hydroxy-, p-methoxy-, p-acetoxy-, and p-cyanobenzyl acetates were highly attractive to the melon fly. Selective fluorination indicated that the polarizability of the carbonyl carbon is important in receptor interaction with the melon fly but is not specifically involved in the oriental fruit fly. Attraction to the melon fly was lost with ortho, meta, or dimethoxy substitution of benzyl acetate, but the 3,4- and 2,5-dimethoxybenzyl acetates were attractive to the oriental fruit fly. These results are interpreted in terms of specific olfactory receptor interactions for the two species of fruit flies.


Increasing Gene Editing Efficiency for CRISPR-Cas9 by Small RNAs in Pluripotent Stem Cells.
Gene manipulations of human induced pluripotent stem cells (iPSCs) by CRISPR-Cas9 genome engineering are widely used for disease modeling and regenerative medicine applications. There are two competing pathways, non-homologous end joining (NHEJ) and homology directed repair (HDR) that correct the double-strand break generated by CRISPR-Cas9. Here, we improved gene editing efficiency of gene knock-in (KI) in iPSCs with minimum components by manipulating the Cas9 expression vector. Either we inserted short hairpin RNA expression cassettes to downregulate DNAPK and XRCC4, two main players of the NHEJ pathway, or we increased cell survival by inserting an anti-apoptotic expression cassette of miRNA-21 into the Cas9 vector. For an easy readout, the pluripotency gene SOX2 was targeted with a T2A-tdTomato reporter construct. In vitro downregulating DNAPK and XRCC4 increased the targeting efficiency of SOX2 KI by around twofold. Furthermore, co-expression of miRNA-21 and Cas9 improved the efficiency of SOX2 KI by around threefold. Altogether, our strategies provide a simple and valuable approach for efficient CRISPR-Cas9 gene editing in iPSCs.


Minimizing false-positives in universal newborn hearing screening: a simple solution.
The false-positive rates of previously reported universal newborn hearing screening (UNHS) programs range between 2.5% and 8%. Critics of UNHS programs have claimed that this rate is too high and might lead to a number of the negative effects produced by false-positive screening tests, namely emotional trauma, disease labeling, iatrogenesis from unnecessary testing, and increased expense in terms of time and money. We previously reported, based on some preliminary data, that as many as 80% of newborns who failed the initial hearing screen subsequently passed when they were retested the following day, before being discharged from the hospital. We now present the results of this intervention for our entire UNHS program during a 7-month period. We analyzed data from 3142 non-neonatal intensive care unit infants screened with an automated auditory brainstem response at the Women's Hospital of Greensboro from November 1, 1999 to May 31, 2000. A protocol was developed wherein all infants who failed the initial UNHS were rescreened with another automated auditory brainstem response before hospital discharge. Data collected included pass/fail rates during the inpatient stay as well as follow-up data and risk factors for congenital hearing loss. Confirmed hearing loss occurred in 8 nonneonatal intensive care unit infants, a rate of 2.5/1000. Eighty percent of newborns who failed the initial hearing screen passed on rescreening before hospital discharge. This produced a false-positive rate of 0.8% and a corresponding positive predictive value of 24%. If inhospital rescreening had not occurred, our false-positive rate and positive predictive value would have been 3.9% and 6.1%, respectively. Our simple intervention of rescreening all infants who failed their initial UNHS before hospital discharge reduced the false-positive rate of UNHS to 0.8%. We suggest that this simple, inexpensive intervention should be instituted for all similar UNHS programs.


Supporting Better Evidence Generation and Use within Social Innovation in Health in Low- and Middle-Income Countries: A Qualitative Study.
While several papers have highlighted a lack of evidence to scale social innovations in health, fewer have explored decision-maker understandings of the relative merit of different types of evidence, how such data are interpreted and applied, and what practical support is required to improve evidence generation. The objectives of this paper are to understand (1) beliefs and attitudes towards the value of and types of evidence in scaling social innovations for health, (2) approaches to evidence generation and evaluation used in systems and policy change, and (3) how better evidence-generation can be undertaken and supported within social innovation in health. Thirty-two one-on-one interviews were conducted between July and November 2015 with purposively selected practitioners, policymakers, and funders from low- and middle- income countries (LMICs). Data were analysed using a Framework Analysis Approach. While practitioners, funders, and policymakers said they held outcome evidence in high regard, their practices only bear out this assertion to varying degrees. Few have given systematic consideration to potential unintended consequences, in particular harm, of the programs they implement, fund, or adopt. Stakeholders suggest that better evidence-generation can be undertaken and supported within social innovation in health by supporting the research efforts of emerging community organizations; creating links between practitioners and academia; altering the funding landscape for evidence-generation; providing responsive technical education; and creating accountability for funders, practitioners, and policymakers. How better evidence-generation can be undertaken and supported within social innovation in health is a previously under-operationalised aspect of the policy-making process that remains essential in order to refrain from causing harm, enable the optimization of existing interventions, and ultimately, to scale and fund what works.


Xer1-mediated site-specific DNA inversions and excisions in Mycoplasma agalactiae.
Surface antigen variation in Mycoplasma agalactiae, the etiologic agent of contagious agalactia in sheep and goats, is governed by site-specific recombination within the vpma multigene locus encoding the Vpma family of variable surface lipoproteins. This high-frequency Vpma phase switching was previously shown to be mediated by a Xer1 recombinase encoded adjacent to the vpma locus. In this study, it was demonstrated in Escherichia coli that the Xer1 recombinase is responsible for catalyzing vpma gene inversions between recombination sites (RS) located in the 5'-untranslated region (UTR) in all six vpma genes, causing cleavage and strand exchange within a 21-bp conserved region that serves as a recognition sequence. It was further shown that the outcome of the site-specific recombination event depends on the orientation of the two vpma RS, as direct or inverted repeats. While recombination between inverted vpma RS led to inversions, recombination between direct repeat vpma RS led to excisions. Using a newly developed excision assay based on the lacZ reporter system, we were able to successfully demonstrate under native conditions that such Xer1-mediated excisions can indeed also occur in the M. agalactiae type strain PG2, whereas they were not observed in the control xer1-disrupted VpmaY phase-locked mutant (PLMY), which lacks Xer1 recombinase. Unless there are specific regulatory mechanisms preventing such excisions, this might be the cost that the pathogen has to render at the population level for maintaining this high-frequency phase variation machinery.


Bevacizumab and platinum-based combinations for recurrent ovarian cancer: a randomised, open-label, phase 3 trial.
State-of-the art therapy for recurrent ovarian cancer suitable for platinum-based re-treatment includes bevacizumab-containing combinations (eg, bevacizumab combined with carboplatin-paclitaxel or carboplatin-gemcitabine) or the most active non-bevacizumab regimen: carboplatin-pegylated liposomal doxorubicin. The aim of this head-to-head trial was to compare a standard bevacizumab-containing regimen versus carboplatin-pegylated liposomal doxorubicin combined with bevacizumab. This multicentre, open-label, randomised, phase 3 trial, was done in 159 academic centres in Germany, France, Australia, Austria, and the UK. Eligible patients (aged ≥18 years) had histologically confirmed epithelial ovarian, primary peritoneal, or fallopian tube carcinoma with first disease recurrence more than 6 months after first-line platinum-based chemotherapy, and an Eastern Cooperative Oncology Group performance status of 0-2. Patients were stratified by platinum-free interval, residual tumour, previous antiangiogenic therapy, and study group language, and were centrally randomly assigned 1:1 using randomly permuted blocks of size two, four, or six to receive six intravenous cycles of bevacizumab (15 mg/kg, day 1) plus carboplatin (area under the concentration curve [AUC] 4, day 1) plus gemcitabine (1000 mg/m2, days 1 and 8) every 3 weeks or six cycles of bevacizumab (10 mg/kg, days 1 and 15) plus carboplatin (AUC 5, day 1) plus pegylated liposomal doxorubicin (30 mg/m2, day 1) every 4 weeks, both followed by maintenance bevacizumab (15 mg/kg every 3 weeks in both groups) until disease progression or unacceptable toxicity. There was no masking in this open-label trial. The primary endpoint was investigator-assessed progression-free survival according to Response Evaluation Criteria in Solid Tumors version 1.1. Efficacy data were analysed in the intention-to-treat population. Safety was analysed in all patients who received at least one dose of study drug. This completed study is registered with ClinicalTrials.gov, NCT01837251. Between Aug 1, 2013, and July 31, 2015, 682 eligible patients were enrolled, of whom 345 were randomly assigned to receive carboplatin-pegylated liposomal doxorubicin-bevacizumab (experimental group) and 337 were randomly assigned to receive carboplatin-gemcitabine-bevacizumab (standard group). Median follow-up for progression-free survival at data cutoff (July 10, 2018) was 12·4 months (IQR 8·3-21·7) in the experimental group and 11·3 months (8·0-18·4) in the standard group. Median progression-free survival was 13·3 months (95% CI 11·7-14·2) in the experimental group versus 11·6 months (11·0-12·7) in the standard group (hazard ratio 0·81, 95% CI 0·68-0·96; p=0·012). The most common grade 3 or 4 adverse events were hypertension (88 [27%] of 332 patients in the experimental group vs 67 [20%] of 329 patients in the standard group) and neutropenia (40 [12%] vs 73 [22%]). Serious adverse events occurred in 33 (10%) of 332 patients in the experimental group and 28 (9%) of 329 in the standard group. Treatment-related deaths occurred in one patient in the experimental group (<1%; large intestine perforation) and two patients in the standard group (1%; one case each of osmotic demyelination syndrome and intracranial haemorrhage). Carboplatin-pegylated liposomal doxorubicin-bevacizumab is a new standard treatment option for platinum-eligible recurrent ovarian cancer. F Hoffmann-La Roche.


Measurement of microchannel flow with digital holographic microscopy by integrated nearest neighbor and cross-correlation particle pairing.
A micro digital in-line holographic particle tracking velocimetry (micro-DHPTV) system has been developed and applied to investigate the three-dimensional flow field in straight and Y-junction microchannels. The micro-DHPTV system comprises a cooled frame-transfer CCD camera and a double-pulsed laser. The processing algorithm introduced to evaluate the three-dimensional velocity is based on the combination of integrated cross-correlation and nearest neighbor matching algorithms, taking advantage of information from both the reconstructed particle field and the original holograms fringes patterns. Tests on simulated pairs of holograms show that the particles can be detected, located, and paired with high probability and accuracy. Results obtained in the straight and Y-junction microchannels show that the superimposed vector field is physically reasonable.


Quantitative investigations of idiotypic antibodies. V. Factors affecting the persistence and replacement of clones of antibody-producing cells.
The effect of challenge by antigen on persistence of clones of antibody-producing cells and on the induction of new clones was investigated through quantitative measurements of idiotypic specificities. In each of nine rabbits idiotypic specificities present in the earliest bleedings were completely replaced after a few months; subsequent changes occurred much more slowly. On a quantitative basis the population of molecules used as immunogen always reacted most effectively with the homologous anti-idiotypic antiserum. Little effect of increased antigen dose on the rate of change of idiotype was observed. Even large amounts of antigen administered every 2 wk caused only gradual changes in idiotypic specificities. This was attributed either to more effective capture of antigen by memory cells, as compared to precursor cells, or to the induction of tolerance in those clones that were not expressed. In two of three rabbits on a monthly injection schedule, the idiotypic specificities identified underwent very slow changes over a period as long as 17 months. Changes occurred more rapidly when antigen was administered every 2 wk. In each of four rabbits investigated, all idiotypic specificities identified before a 5 month rest period were still present afterwards, indicating the survival of essentially all clones of antibody-producing cells during that interval. Quantitative inhibition data indicated that some new clones of cells were initiated.


Efficiency of recombinant human TNF in human cancer therapy.
Recombinant human tumour necrosis factor (TNF) has a selective effect on angiogenic vessels in tumours. Given that it induces vasoplegia, its clinical use has been limited to administration through isolated limb perfusion (ILP) for regionally advanced melanomas and soft tissue sarcomas of the limbs. When combined with the alkylating agent melphalan, a single ILP produces a very high objective response rate. In melanoma, the complete response (CR) rate is around 80% and the overall objective response rate greater than 90%. In soft tissue sarcomas that are inextirpable, ILP is a neoadjuvant treatment resulting in limb salvage in 80% of the cases. The CR rate averages 20% and the objective response rate is around 80%. The mode of action of TNF-based ILP involves two distinct and successive effects on the tumour-associated vasculature: first, an increase in endothelium permeability leading to improved chemotherapy penetration within the tumour tissue, and second, a selective killing of angiogenic endothelial cells resulting in tumour vessel destruction. The mechanism whereby these events occur involves rapid (of the order of minutes) perturbation of cell-cell adhesive junctions and inhibition of alphavbeta3 integrin signalling in tumour-associated vessels, followed by massive death of endothelial cells and tumour vascular collapse 24 hours later. New, promising approaches for the systemic use of TNF in cancer therapy include TNF targeting by means of single chain antibodies or endothelial cell ligands, or combined administration with drugs perturbing integrin-dependent signalling and sensitizing angiogenic endothelial cells to TNF-induced death.


Optimising a product standard for banning menthol and other flavours in tobacco products.
In this paper, we highlight key issues that policymakers should consider when developing a product standard banning menthol and other flavours in tobacco products based on research evidence and experiences learnt from other countries. A flavour product standard may be optimised by (1) having a clear and comprehensive definition of flavour that includes a complete ban on additives that have flavour properties and/or evoke sensory/cooling effects (ie, menthol analogues and synthetic coolants that stimulate the cooling receptor of the brain) rather than only as a 'characterising flavour' and (2) applying the standard to all tobacco product categories as well as all components or parts of the tobacco product (ie, the tobacco, filter, wrapper or paper), including separate flavourings that can be added to the product.


Characterization of a novel adeno-associated viral vector with preferential oligodendrocyte tropism.
No adeno-associated virus (AAV) capsid has been described in the literature to exhibit a primary oligodendrocyte tropism when a constitutive promoter drives gene expression, which is a significant barrier for efficient in vivo oligodendrocyte gene transfer. The vast majority of AAV vectors, such as AAV1, 2, 5, 6, 8 or 9, exhibit a dominant neuronal tropism in the central nervous system. However, a novel AAV capsid (Olig001) generated using capsid shuffling and directed evolution was recovered after rat intravenous delivery and subsequent capsid clone rescue, which exhibited a >95% tropism for striatal oligodendrocytes after rat intracranial infusion where a constitutive promoter drove gene expression. Olig001 contains a chimeric mixture of AAV1, 2, 6, 8 and 9, but unlike these parental serotypes after intravenous administration Olig001 has very low affinity for peripheral organs, especially the liver. Furthermore, in mixed glial cell cultures, Olig001 exhibits a 9-fold greater binding when compared with AAV8. This novel oligodendrocyte-preferring AAV vector exhibits characteristics that are a marked departure from previously described AAV serotypes.


Biopolymer-induced calcium phosphate scaling in membrane-based water treatment systems: Langmuir model films studies.
Biofouling and scaling on reverse osmosis (RO) or nanofiltration (NF) membranes during desalination of secondary and tertiary effluents pose an obstacle that limits the reuse of wastewater. In this study we explored the mineral scaling induced by biopolymers originated from bacterial biofilms: bovine serum albumin (BSA), fibrinogen, lysozyme and alginic acid, as well as an extracts of extracellular polymeric substances (EPS) from bio-fouled RO membranes from wastewater treatment facility. Mineralization studies were performed on Langmuir films of the biopolymers deposited at the interface of a solution simulating RO desalination of secondary-treated wastewater effluents. All studied biopolymers and EPS induced heterogeneous mineralization of mainly calcium phosphate. Using IR spectroscopy coupled with systematic quantitative analysis of the surface pressure versus molecular-area isotherms, we determined the mineralization tendencies of the biopolymers to be in the order of: fibrinogen>lysozyme>BSA>alginic acid. The biopolymers and EPS studied here were found to be accelerators of calcium-phosphate mineralization. This study demonstrates the utilization of Langmuir surface-pressure area isotherms and a model solution in quantitatively assessing the mineralization tendencies of various molecular components of EPS in context of membrane-based water treatment systems.


Study design implications of death and hospitalization as end points in idiopathic pulmonary fibrosis.
The feasibility of an interventional clinical trial in idiopathic pulmonary fibrosis (IPF) using death and hospitalization as primary end points is an area of uncertainty. Using data from a large well-characterized clinical trial population, this article aims to illustrate the impact of cohort enrichment and study duration on sample size requirements for IPF clinical trials in which death alone or death plus hospitalization serve as the primary end point. Event rate estimates for death and hospitalization were determined from patients enrolled in National Institutes of Health-sponsored IPF Clinical Research Network clinical trials. Standard equations were applied to estimate the total sample size required for varying gender, age, and pulmonary function (GAP) stage-based cohorts. Risk estimates for death and hospitalization in the clinical trial cohort were substantially lower than those published. An IPF trial with death as its primary end point enrolling subjects designated as GAP stage 1 and 2 over 1 year with a minimum follow-up of 1 year would require an estimated 7,986 subjects to achieve 90% power for a hazard ratio of 0.70. Alternatively, an IPF trial with death plus hospitalization as its primary end point enrolling subjects with GAP stage 2 and 3 over 2 years with a minimum follow-up of 1 year would require an estimated 794 subjects for the same power and hazard ratio. Study design decisions, in particular cohort enrichment strategies, have a substantial impact on sample size requirements for IPF clinical trials using time-to-event primary end points such as death and death plus hospitalization.


A prospectively randomized double-blind study on the effect of initial graft tension on knee stability after anterior cruciate ligament reconstruction.
No consensus exists on the amount of tension that should be applied to anterior cruciate ligament grafts to best facilitate graft incorporation and re-create normal knee mechanics. Differences in initial graft tension will affect postoperative knee stability. Prospective, randomized, double-blind clinical trial. Forty-nine patients undergoing bone-patellar tendon-bone autograft anterior cruciate ligament reconstruction by a single surgeon were randomized into high-tension (n = 27) and low-tension (n = 22) groups. Grafts were set at 90 N or 45 N. Arthrometric measurements (KT-1000 arthrometer manual maximum) of anterior tibial displacement and knee range of motion were made before surgery and at 1 week and an average of 20 months after surgery. Knee outcome scores were collected before and after surgery, and a single-leg hop test was also performed at final follow-up. After anterior cruciate ligament reconstruction, anterior tibial displacement was significantly greater in the patients in the low-tension group (P < .05). The side-to-side difference in anterior tibial displacement in the high-tension and low-tension groups was 1.1 +/- 1.7 mm versus 2.4 +/- 2.4 mm 1 week after surgery and 2.2 +/- 1.6 mm versus 3.0 +/- 2.2 mm at follow-up. Five patients had abnormal anterior tibial displacement (>5 mm side-to-side difference), and all were in the low-tension group (P <.05). Knee outcome scores improved with surgery (P <.01), with similar results for low-tension and high-tension groups. Hop test deficits were not different between groups. Initial graft tension affects the restoration of knee stability. A graft tension of 45 N was not sufficient for restoring knee stability.


Hepatic Vascular Variants in Hereditary Haemorrhagic Telangiectasia: Imaging findings.
Hereditary haemorrhagic telangiectasia (HHT) is an autosomal dominant disorder characterised by vascular dysplasia. Hepatic vascular malformations (VMs) range from small telangiectases to significant vascular shunting. Here we report two cases of HHT. Case 1 had diffuse ectasia of the hepatic artery along its intrahepatic and extrahepatic course with a hepatic arterial aneurysm. Case 2 presented with ileal and hepatic telangiectases. Knowledge of these vascular variants is indispensable for clinicians and radiologists in aiding diagnosis and surgical and interventional management.


Timed repetitive ankle jerk responses in early diabetic neuropathy.
A 14-year experience with timed repetitive ankle jerk (TRAJ) testing provides fundamental insights into diabetic neuropathy (DN), both its diagnosis and treatment. Advantages of TRAJ include ease of performance, relative economy of the equipment, reproducibility of results, and a parameter that encompasses the entire reflex arc. Euthyroid patients with features suggesting DN often have prolongation of ankle jerk responses on one or both sides only after repetitive stimulation. Prolongation of TRAJ responses were elicited from 41% of overt diabetics and from 27% and 30% of patients with decreased glucose tolerance demonstrated by morning or afternoon testing, respectively. This characteristic response appears to reflect altered neuromuscular function in DN and was not encountered in patients with myasthenia gravis or thyroid dysfunction in the absence of concomitant glucose metabolic aberrations.


Illuminating Clustered Mutations in Cancer.
In mapping the landscape of clustered somatic mutations, researchers have found that such clusters are enriched on driver oncogenes and associated with differential survival. They also reported the prevalence of a particular cluster pattern, kataegis, on extrachromosomal DNA, describing it as kyklonas, potentially a new mode of cancer evolution.


Adherence to the iDSI reference case among published cost-per-DALY averted studies.
The iDSI reference case, originally published in 2014, aims to improve the quality and comparability of cost-effectiveness analyses (CEA). This study assesses whether the development of the guideline is associated with an improvement in methodological and reporting practices for CEAs using disability-adjusted life-years (DALYs). We analyzed the Tufts Medical Center Global Health CEA Registry to identify cost-per-DALY averted studies published from 2011 to 2017. Among each of 11 principles in the iDSI reference case, we translated all methodological specifications and reporting standards into a series of binary questions (satisfied or not satisfied) and awarded articles one point for each item satisfied. We then calculated methodological and reporting adherence scores separately as a percentage of total possible points, measured as normalized adherence score (0% = no adherence; 100% = full adherence). Using the year 2014 as the dissemination period, we conducted a pre-post analysis. We also conducted sensitivity analyses using: 1) optional criteria in scoring, 2) alternate dissemination period (2014-2015), and 3) alternative comparator classification. Articles averaged 60% adherence to methodological specifications and 74% adherence to reporting standards. While methodological adherence scores did not significantly improve (59% pre-2014 vs. 60% post-2014, p = 0.53), reporting adherence scores increased slightly over time (72% pre-2014 vs. 75% post-2014, p<0.01). Overall, reporting adherence scores exceeded methodological adherence scores (74% vs. 60%, p<0.001). Articles seldom addressed budget impact (9% reporting, 10% methodological) or equity (7% reporting, 7% methodological). The iDSI reference case has substantial potential to serve as a useful resource for researchers and policy-makers in global health settings, but greater effort to promote adherence and awareness is needed to achieve its potential.


Allograft inflammatory factor-1 supports macrophage survival and efferocytosis and limits necrosis in atherosclerotic plaques.
Allograft inflammatory factor-1 (AIF1) has been characterized as a pro-inflammatory molecule expressed primarily in the monocyte/macrophage (MP) lineage and positively associated with various forms of vascular disease, including atherosclerosis. Studies of AIF1 in atherosclerosis have relied on mouse models in which AIF1 was overexpressed in either myeloid or smooth muscle cells, resulting in increased atherosclerotic plaque burden. How physiologic expression of AIF1 contributes to MP biology in atherogenesis is not known. Effects of global AIF1 deficiency on atherosclerosis were assessed by crossing Aif1-/- and ApoE-/- mice, and provoking hyperlipidemia with high fat diet feeding. Atherosclerotic plaques were studied en face and in cross section. Bone marrow-derived MPs (BMDMs) were isolated from Aif1-/- mice for study in culture. Atherosclerotic plaques in Aif1-/-;ApoE-/- mice showed larger necrotic cores compared to those in ApoE-/- animals, without change in overall lesion burden. In vitro, lack of AIF1 reduced BMDM survival, phagocytosis, and efferocytosis. Mechanistically, AIF1 supported activation of the NF-κB pathway and expression of related target genes involved in stress response, inflammation, and apoptosis. Consistent with this in vitro BMDM phenotype, AIF1 deficiency reduced NF-κB pathway activity in vivo and increased apoptotic cell number in atherosclerotic lesions from Aif1-/-;ApoE-/- mice. These findings characterize AIF1 as a positive regulator of the NF-κB pathway that supports MP functions such as survival and efferocytosis. In inflammatory settings such as atherosclerosis, these AIF1-dependent activities serve to clear cellular and other debris and limit necrotic core expansion, and may oppose lesion destabilization.


Genetic structure and transforming sequence of avian sarcoma virus UR2.
We have recently shown that a newly isolated avian sarcoma virus, UR2, is defective in replication and contains no sequences homologous to the src gene of Rous sarcoma virus. In this study, we analyzed the genetic structure and transforming sequence of UR2 by oligonucleotide fingerprinting. The sizes of the genomic RNAs of UR2 and its associated helper virus, UR2AV, were determined to be 24S and 35S, respectively, by sucrose gradient sedimentation. The molecular weight of the 24S UR2 genomic RNA was estimated to be 1.1 x 10(6), corresponding to 3,300 nucleotides, by gel electrophoresis under the native and denatured conditions. RNase T1 oligonucleotide mapping indicated that UR2 RNA contains seven unique oligonucleotides in the middle of the genome and shares eight 5'- and six 3'-terminal oligonucleotides with UR2AV RNA. From these data, we estimated that UR2 RNA contains a unique sequence of about 12 kilobases in the middle of the genome, and contains 1.4 and 0.7 kilobases of sequences shared with UR2AV RNA at the 5' and 3' ends, respectively. Partial sequence analysis of the UR2-specific oligonucleotides by RNase A digestion revealed that there are no homologous counterparts to these oligonucleotides in the RNAs of other avian sarcoma and acute leukemia viruses studied to date. UR2-transformed non-virus-producing cells contain a single 24S viral RNA which is most likely the message coding for the transforming protein of UR2. On the basis of the uniqueness of the transforming sequence, we concluded that UR2 is a new member of the defective avian sarcoma viruses.


Targeting Aggressive B-cell Lymphomas through Pharmacological Activation of the Mitochondrial Protease OMA1.
DLBCL are aggressive, rapidly proliferating tumors that critically depend on the ATF4-mediated integrated stress response (ISR) to adapt to stress caused by uncontrolled growth, such as hypoxia, amino acid deprivation, and accumulation of misfolded proteins. Here, we show that ISR hyperactivation is a targetable liability in DLBCL. We describe a novel class of compounds represented by BTM-3528 and BTM-3566, which activate the ISR through the mitochondrial protease OMA1. Treatment of tumor cells with compound leads to OMA1-dependent cleavage of DELE1 and OPA1, mitochondrial fragmentation, activation of the eIF2α-kinase HRI, cell growth arrest, and apoptosis. Activation of OMA1 by BTM-3528 and BTM-3566 is mechanistically distinct from inhibitors of mitochondrial electron transport, as the compounds induce OMA1 activity in the absence of acute changes in respiration. We further identify the mitochondrial protein FAM210B as a negative regulator of BTM-3528 and BTM-3566 activity. Overexpression of FAM210B prevents both OMA1 activation and apoptosis. Notably, FAM210B expression is nearly absent in healthy germinal center B-lymphocytes and in derived B-cell malignancies, revealing a fundamental molecular vulnerability which is targeted by BTM compounds. Both compounds induce rapid apoptosis across diverse DLBCL lines derived from activated B-cell, germinal center B-cell, and MYC-rearranged lymphomas. Once-daily oral dosing of BTM-3566 resulted in complete regression of xenografted human DLBCL SU-DHL-10 cells and complete regression in 6 of 9 DLBCL patient-derived xenografts. BTM-3566 represents a first-of-its kind approach of selectively hyperactivating the mitochondrial ISR for treating DLBCL.


Non-Parental Adults in the Social and Risk Behavior Networks of Sexual Minority Male Youth.
The presence of non-parental adults (NPAs), or adults outside of caregivers (e.g., extended family, natural mentors), in the lives of adolescents and emerging adults has received a rapidly expanding amount of empirical attention in the last decade. Sexual minority male youth (SMMY) face disproportionate risks of abuse and victimization in relationships with parents and peers. Yet, despite the fact that this group, therefore, may be both potentially vulnerable to negative interpersonal influences but also poised to benefit from additional relationships, NPA involvement in the lives of SMMY is currently not well understood in the extant literature. This study sought to examine and characterize the involvement of NPAs in the social and risk networks of SMMY (n = 175; 54% African American, 21% Hispanic/Latino, 14% Caucasian; ages 17-23). Most SMMY identified at least one NPA, such as friends and grandparents, in their networks. Three categories of relationships were identified, Strictly Social, which only involved social interactions; Complex, which were both social and involved substance use and/or sexual activity; and Risky, which purely consisted of substance use or sexual activity. Relationships were rated as emotionally "closer" among ethnic minority SMMY, although, racial/ethnic similarity between SMMY and NPAs was not associated with relationship closeness. In addition, relationships involving female and heterosexual NPAs were also rated as stronger. These findings suggest the potential usefulness of considering multiple types of relationships between SMMY and NPAs when designing intervention and prevention efforts. Moreover, African American and Latino SMMY, who represent the most vulnerable sub-groups of SMMY in terms of HIV-risk, may be particularly poised to benefit from positive NPA relationships.


Exon-intron structure of a 2.7-kb transcript of the STM7 gene with phosphatidylinositol-4-phosphate 5-kinase activity.
The STM7 gene encodes a novel phosphatidylinositol-4-phosphate 5-kinase (PtdInsP 5-kinase) that is subject to alternative splicing and developmental control. We have recently presented data indicating that several splice variants of STM7 incorporate elements of the X25 sequence, previously implicated in the pathogenesis of Friedreich's ataxia by the detection of an intronic GAA repeat expansion as the predominant mutation in affected individuals. We now report the exon-intron structure of STM7.I and primer sequences designed to facilitate full characterization, including details relating to a novel exon (STM7; exon 17) derived from the 3'-UTR of the PRKACG gene. The detection of a mutation(s) within these exons would provide additional support for the hypothesis that a defect in phosphoinositide metabolism gives rise to the disease phenotype.


LAPTM4B-35, a cancer-related gene, is associated with poor prognosis in TNM stages I-III gastric cancer patients.
Lysosome-associated transmembrane protein 4β-35 (LAPTM4B-35), a member of the mammalian 4-tetratransmembrane spanning protein superfamily, has been reported to be overexpressed in several cancers. However the expression of LAPTM4B-35 and its role in the progression of gastric cancer (GC) remains unknown. The aim of this study was to investigate LAPTM4B-35 expression in GC, its potential relevance to clinicopathologic parameters and role of LAPTM4B-35 during gastric carcinogenesis. In the present study, paraffin-embedded specimens with GC (n = 240, including 180 paired specimens) and 24 paired fresh frozen tissues were analyzed. qRT-PCR and immunohistochemistry (IHC) were used to analyze the expression of LAPTM4B-35 in GC. The effects of LAPTM4B-35 on GC cell proliferation, migration and invasion were determined by overexpression and knockdown assays. IHC showed that LAPTM4B-35 was expressed in 68.3% (123/180) of GC tissues, while in 16.1% (29/180) of their paired adjacent noncancerous gastric tissues (P = 0.000). LAPTM4B-35 mRNA levels in GC tissues were also significantly elevated when compared with their paired adjacent noncancerous tissues (P = 0.017). Overexpression of LAPTM4B-35 was significantly associated with degree of differentiation, depth of invasion, lymphovascular invasion and lymph node metastasis (P<0.05). Kaplan-Meier survival curves revealed that patients with LAPTM4B-35 expression had a significant decrease in overall survival (OS) in stages I-III GC patients (P = 0.006). Multivariate analysis showed high expression of LAPTM4B-35 was an independent prognostic factor for OS in stage I-III GC patients (P = 0.025). These findings indicate that LAPTM4B-35 overexpression may be related to GC progression and poor prognosis, and thus may serve as a new prediction marker of prognosis in GC patients.


pH-Responsive polyelectrolyte coated gadolinium oxide-doped mesoporous silica nanoparticles (Gd2O3@MSNs) for synergistic drug delivery and magnetic resonance imaging enhancement.
Theranostic platforms that combine therapeutic and imaging modalities have received increasing interest. The development of theranostic nanovectors that can release therapeutic agents at pathological tissues in an on-demand manner and provide instantaneous feedback through non-invasive imaging techniques is of urgent need. Herein, a new magnetic resonance imaging (MRI) contrast agent, gadolinium oxide (Gd2O3), and an anticancer drug, doxorubicin (DOX), were co-loaded into mesoporous silica nanoparticles (MSNs) with the formation of hybrid Gd2O3@MSN-DOX nanoparticles. The hybrid nanoparticles were further coated with pH-responsive polyelectrolytes that underwent a charge reversal process at acidic pH. Upon entering into cells by folic acid (FA) receptor-mediated endocytosis, the mildly acidic pH within endolysosomes triggered the disassociation of the absorbed polyelectrolytes on the surfaces of the Gd2O3@MSN-DOX nanoparticles and thus actuated the DOX release, thereby exerting an anti-cancer effect. More importantly, the confinement of paramagnetic Gd2O3 within MSNs led to a remarkable increase of MRI relaxivity (r1 = 9.14 mM-1 s-1vs. 3.68 mM-1 s-1 of the clinically applied MRI contrast agent), likely due to the increased tumbling time and coordination number of water molecules. This work provides a feasible strategy to fabricate theranostic nanovectors with controlled release behavior triggered by mildly acidic pH and high-performance MR imaging capability.


Prevalence of pulmonary arterial hypertension in the Camerino area of central Italy and savings resulting from generic bosentan.
Pulmonary arterial hypertension is a rare and progressive respiratory disease characterised by high blood pressure and vascular resistance producing right ventricular fatigue. In Italy, pulmonary hypertension can be treated with different drugs available on the market at different costs, and in the Marche region distributed exclusively by hospital pharmacies. The present study examined in an area of the Marche region the use of drugs specifically indicated for pulmonary hypertension, and evaluated how the introduction of the generic bosentan might lower pharmaceutical costs for the healthcare budget. The study examined oral administration prescriptions and costs using data from the Apotheke Gold (Record Data) database from 1 January 2012 to 31 August 2017. Annually (from 1 January 2012 to 31 August 2017), an average of 4.83 patients were treated (prevalence of 102.35 cases per 1 million residents) with ambrisentan (Volibris), bosentan (Tracleer), macitentan (Opsumit), tadalafil (Adcirca) or sildenafil (Revatio). The total expenditure during the 5-year 8-month period was €472 405. Ambrisentan was by far the most expensive product overall, with a total expenditure of €222 380 for the period studied (a daily cost of €67.39), even though Tracleer had the highest cost for a day of treatment (a daily cost of €94.48, but a total expenditure of €163 976 for the period, due to its more recent marketing). Providing patients with the generic form bosentan in place of Tracleer would lower the costs dramatically. A very significant annual savings per patient of approximately €31 879 would be achieved, a striking 92.4% reduction in costs. The prevalence of pulmonary arterial hypertension reported for Camerino and its surrounding area in the Marches region is quite high compared with that reported by other authors for France and Scotland. The introduction of the generic bosentan would cut costs drastically. It is to be hoped that centralised procurement at the regional level would bring further savings.


Circular RNAs in Muscle Function and Disease.
Circular RNAs (circRNAs) are a class of RNA produced during pre-mRNA splicing that are emerging as new members of the gene regulatory network. In addition to being spliced in a linear fashion, exons of pre-mRNAs can be circularized by use of the 3' acceptor splice site of upstream exons, leading to the formation of circular RNA species. In this way, genetic information can be re-organized, increasing gene expression potential. Expression of circRNAs is developmentally regulated, tissue and cell-type specific, and shared across eukaryotes. The importance of circRNAs in gene regulation is now beginning to be recognized and some putative functions have been assigned to them, such as the sequestration of microRNAs or proteins, the modulation of transcription, the interference with splicing, and translation of small proteins. In accordance with an important role in normal cell biology, circRNA deregulation has been reported to be associated with diseases. Recent evidence demonstrated that circRNAs are highly expressed in striated muscle tissue, both skeletal and cardiac, that is also one of the body tissue showing the highest levels of alternative splicing. Moreover, initial studies revealed altered circRNA expression in diseases involving striated muscle, suggesting important functions of these molecules in the pathogenetic mechanisms of both heart and skeletal muscle diseases. The recent findings in this field will be described and discussed.


Maternal hepatitis B or hepatitis C virus carrier status and long-term infectious morbidity of the offspring: A population-based cohort study.
The objective of the study was to investigate the long-term effects of maternal hepatitis B virus (HBV) or hepatitis C virus (HCV) carrier status on the long-term infectious morbidity of their offspring. A population-based cohort study was conducted, including all singleton deliveries between the years 1991 and 2014 at a tertiary medical centre. The mothers were subdivided into three groups: HBV carriers, HCV carriers and non-carriers. Data on demographics, maternal, perinatal and long-term hospitalization for infectious morbidity were compared between the groups. During the study period, 242 905 (99.7%) non-carrier mothers, 591 (0.2%) HBV carriers and 186 (0.1%) HCV carriers were observed. Hospitalizations related to infectious morbidity was significantly higher in the offspring of HBV carriers compared with HCV and non-carriers (15.6% vs 11.3% vs 11.0%; P = .002, respectively; Kaplan-Meier, log-rank P < .001). Specifically, a significantly higher rate of hospitalizations gastrointestinal infectious morbidity was noted among the offspring of HBV carrier mothers (3.6% in the HBV carrier group, 1.6% in the HCV carrier group and 1.6% in the non-carrier group [P = .001]). There was a respiratory infectious morbidity of 8.1% among the offspring of HBV carriers, 8.6% among HCV carriers and 5.5% in non-carriers (P = .005). Using a Cox multivariable model, controlling for confounding variables, maternal HBV carrier status was associated with a significantly increased long-term infectious morbidity of the offspring, with an adjusted HR of 1.7 (95% CI, 1.388-2.077, P < .001). Maternal HBV carrier status is an independent risk factor for long-term infectious morbidity of the offspring, particularly for gastrointestinal and respiratory infections.


Characterization of the human p53 gene.
Cosmid and lambda clones containing the human p53 gene were isolated and characterized in detail. The gene is 20 kilobases (kb) long and has 11 exons, the first and second exons being separated by an intron of 10 kb. Restriction fragments upstream of sequences known to be within the first identified exon were tested for promoter activity by cloning them in front of the chloramphenicol acetyltransferase gene and transfecting the resulting constructs into HeLa cells. A 0.35-kb DNA fragment was identified that had promoter activity. Results of primer extension experiments indicated that the mRNA cap site falls within this fragment, as expected. Analysis of the sequence upstream of the presumptive cap site indicated that the human p53 promoter may be of an unusual type.


Flow cytometric analysis of adherence of Porphyromonas gingivalis to oral epithelial cells.
By using fluorescence microscopy, fluorescently labeled Porphyromonas gingivalis W50 was shown to adhere to oral epithelial (KB) cells as discrete cells or small cell aggregates, whereas P. gingivalis ATCC 33277 bound as large cell aggregates. Flow cytometric analysis showed that for P. gingivalis W50 there was a logarithmic relationship between the bacterial cell ratio (BCR), that is the number of bacterial cells to KB cells, and the percentage of KB cells with W50 cells attached. This percentage of KB cells with W50 attached reached a plateau of approximately 84% cells at a BCR of 500:1. In contrast, a quadratic relationship was observed between BCR and the percentage of KB cells with P. gingivalis ATCC 33277 attached, reaching a maximum of 47% at a BCR of 100:1 but decreasing to 7% at a BCR of 1,000:1. The lower binding of ATCC 33277 at high cell concentrations was attributed to autoaggregation. P. gingivalis W50 cells treated with an inhibitor (Nalpha-p-tosyl-L-lysine chloromethyl ketone [TLCK]) of its RgpA-Kgp proteinase-adhesin complex exhibited significantly reduced binding to KB cells than to untreated cells, suggesting a role for proteinase activity in binding to KB cells. Competitive inhibition with purified proteinase-active and TLCK-inactivated RgpA-Kgp complex significantly decreased the adherence of P. gingivalis W50 cells to KB cells. Furthermore, isogenic mutants of P. gingivalis W50 lacking the kgp gene product, but not the rgpA or rgpB gene products, exhibited significantly decreased adherence to KB cells compared to the wild type.


The role of molecular imaging in diagnosis of deep vein thrombosis.
Venous thromboembolism (VTE) mostly presenting as deep venous thrombosis (DVT) and pulmonary embolism (PE) affects up to 600,000 individuals in United States each year. Clinical symptoms of VTE are nonspecific and sometimes misleading. Additionally, side effects of available treatment plans for DVT are significant. Therefore, medical imaging plays a crucial role in proper diagnosis and avoidance from over/under diagnosis, which exposes the patient to risk. In addition to conventional structural imaging modalities, such as ultrasonography and computed tomography, molecular imaging with different tracers have been studied for diagnosis of DVT. In this review we will discuss currently available and newly evolving targets and tracers for detection of DVT using molecular imaging methods.


A summary of knowledge about the oral health of older people in England and Wales.
To summarise what is currently known about the oral health of older adults in England and Wales. Summary of the main findings from a recent review of oral health surveys and demographic and health data relating to older people in the UK (West Midlands, North West, Bolton and Kirklees, East London and the City of London and Wales). Their findings were compared, where possible to estimates from the 2009 Adult Dental Health Survey. A higher proportion of older adults in England and Wales have untreated caries and signs of severe caries than the general adult population. The majority of dentate residents in the care homes surveyed had untreated caries. Despite the poorer oral health of residents in care homes, managers of such services report difficulty in accessing routine and emergency dental care. Existing epidemiological data in England and Wales show that older people in residential and nursing care homes have poorer oral health than the general adult population and inequitable access to dental services. Greater comparability and utility would be gained from regional oral health surveys if standards were agreed for this age group with regard to sampling, consent, questionnaires and clinical measures.


A palmtop computer program for the treatment of generalized anxiety disorder.
This is the first report of a palmtop computer program developed to increase the efficiency and cost-effectiveness of cognitive behavioral therapy for generalized anxiety disorder (GAD). The computer program offers advantages to researchers, therapists, and clients. These advantages include continuous, unobtrusive collection of process data on treatment adherence as well as on the impact of cognitive behavioral therapy techniques in the client's natural setting. In addition, the computer extends treatment beyond the therapy hour and motivates clients to comply with homework assignments by prompting practice of cognitive behavioral strategies. The successful application of the palmtop computer program reported in this integrated series suggests a new line of research directed toward increasing the cost-effectiveness of what is currently the gold-standard treatment for GAD.


Local application of melatonin into alveolar sockets of beagle dogs reduces tooth removal-induced oxidative stress.
The antioxidant and anti-inflammatory hormone melatonin is secreted by saliva into the oral cavity, where it may protect the mucosal and gingival tissues from radical damage. To date, no studies have addressed the potential beneficial role of melatonin in the acute inflammatory response that follows oral surgical interventions, especially tooth extractions. The aim of this study was to determine whether tooth extraction induces changes in plasma oxidative stress levels, and whether melatonin treatment may counteract these changes. Maxillary and mandibular premolars and molars of 16 adult Beagle dogs were extracted under general anesthesia. Eight dogs were treated with 2 mg melatonin placed into the alveolar sockets, whereas the other eight dogs received only vehicle. Lipid peroxidation (LPO) and nitrite plus nitrate (NOx) levels were determined in plasma, whereas glutathione (GSH) and glutathione disulfide (GSSG) levels and glutathione peroxidase (GPx) and reductase (GRd) activities were measured in red blood cells before and 24 hours after tooth extraction. Removal of the premolars and molars caused a significant rise in plasma LPO and NOx levels and in the erythrocyte GSSG/GSH ratio, whereas melatonin treatment restored the normal values of these parameters. Also, melatonin slightly increased erythrocyte GRd activity without changing GPx activity. For the first time to our knowledge, the results show that during the immediate postoperative period following tooth extraction, there is a significant increase of oxidative stress, which is counteracted by the administration of melatonin into the alveolar sockets.


Randomized trial of supplemental beta-carotene to prevent second head and neck cancer.
Beta-carotene has established efficacy in animal models of oral carcinogenesis and has been shown to regress oral precancerous lesions in humans. The purpose of this study was to see whether these effects extended to the prevention of oral/pharyngeal/laryngeal (head and neck) cancer in humans. The subject population for this randomized, placebo-controlled, double-blinded clinical trial included 264 patients who had been curatively treated for a recent early-stage squamous cell carcinoma of the oral cavity, pharynx, or larynx. Patients were assigned randomly to receive 50 mg of beta-carotene per day or placebo and were followed for up to 90 months for the development of second primary tumors and local recurrences. After a median follow-up of 51 months, there was no difference between the two groups in the time to failure [second primary tumors plus local recurrences: relative risk (RR), 0.90; 95% confidence interval (CI), 0.56-1.45]. In site-specific analyses, supplemental beta-carotene had no significant effect on second head and neck cancer (RR, 0.69; 95% CI, 0.39-1.25) or lung cancer (RR, 1.44; 95% CI, 0.62-3.39). Total mortality was not significantly affected by this intervention (RR, 0.86; 95% CI, 0.52-1.42). Whereas none of the effects were statistically significant, the point estimates suggested a possible decrease in second head and neck cancer risk but a possible increase in lung cancer risk. These effects are consistent with the effects observed in trials using intermediate end point biological markers in humans, in which beta-carotene has established efficacy in oral precancerous lesions but has no effect or slightly worsens sputum cytology, and in animal carcinogenicity studies, in which beta-carotene has established efficacy in buccal pouch carcinogenesis in hamsters but not in animal models of respiratory tract/lung carcinogenesis, with some suggestions of tumor-promoting effects in respiratory tract/lung. If our results are replicated by other ongoing/completed trials, this suggests a critical need for mechanistic studies addressing differential responses in one epithelial site (head and neck) versus another (lung).


Effect of magnesium sulfate on oxygenation and lung mechanics in morbidly obese patients undergoing bariatric surgery: a prospective double-blind randomized clinical trial.
Respiratory mechanics are often significantly altered in morbidly obese patients and magnesium sulfate (MgSO4) is a promising agent for managing several respiratory disorders. This study aimed to examine the effects of MgSO4 infusions on arterial oxygenation and lung mechanics in patients with morbid obesity undergoing laparoscopic bariatric surgery. Forty patients with morbid obesity aged 21-60 years scheduled for laparoscopic bariatric surgery under general anesthesia were randomly allocated to either the control (normal saline infusion) or MgSO4 group (30 mg/kg lean body weight [LBW] of 10% MgSO4 in 100 ml normal saline intravenously over 30 min as a loading dose, followed by 10 mg/kg LBW/h for 90 min). The primary outcome was intraoperative arterial oxygenation (ΔPaO2/FiO2). Secondary outcomes included intraoperative static and dynamic compliance, dead space, and hemodynamic parameters. At 90 min intraoperatively, the Δ PaO2/FiO2 ratio and the Δ dynamic lung compliance were statistically significantly higher in the MgSO4 group (mean ± SE: 16.1 ± 1.0, 95% CI [14.1, 18.1] and 8.4 ± 0.5 ml/cmH2O, 95% CI [7.4, 9.4]), respectively), and the Δ dead space (%) was statistically significantly lower in the MgSO4 group (mean ± SE: -8.0 ± 0.3%, 95% CI [-8.6, -7.4]) (P < 0.001). No significant differences in static compliance were observed. Although MgSO4 significantly preserved arterial oxygenation and maintained dynamic lung compliance and dead space in patients with morbid obesity, the clinical relevance is minimal. This study failed to adequately reflect the clinical importance of these results.


Differences in the prevalence and severity of arthritis among racial/ethnic groups in the United States, National Health Interview Survey, 2002, 2003, and 2006.
We describe the prevalence of doctor-diagnosed arthritis and its impact on activities, work, and joint pain for 6 racial/ethnic groups: non-Hispanic whites, non-Hispanic blacks, Hispanics, American Indians/Alaska Natives, Asians and Pacific Islanders, and multiracial or "other" respondents. We combined data from the 2002, 2003, and 2006 National Health Interview Survey (n = 85,784) and, after adjusting for age, sex, and body mass index, compared racial/ethnic differences. Arthritis-attributable activity limitation, arthritis-attributable work limitation, and severe joint pain were higher for non-Hispanic blacks, Hispanics, and multiracial or other respondents with arthritis compared with non-Hispanic whites with arthritis. Our finding that arthritis disproportionately affects certain racial/ethnic minorities may be useful for planning interventions.


Association between plantar fascia vascularity and morphology and foot dysfunction in individuals with chronic plantar fasciitis.
Single-cohort laboratory-based study. To identify whether plantar fascia vascularity and thickness are associated with foot pain and dysfunction in individuals with chronic plantar fasciitis. Background Altered plantar fascia vascularity and thickening of the fascia have been identified in individuals with chronic plantar fasciitis. Thirty-eight patients with chronic unilateral plantar fasciitis and 21 controls participated in this study. Proximal plantar fascia vascularization and thickness were assessed using ultrasound imaging, and pain and foot dysfunction were quantified with a visual analog scale and the Chinese version of the Foot Function Index, respectively. Paired t tests were used to assess the side-to-side differences in fascia thickness and vascularity index (VI) in the control and patient groups, and an unpaired t test was used to make comparisons with the patient group. Multiple regression analysis was performed to identify whether the VI and fascia thickness were associated with pain and foot dysfunction. There were significantly higher VI (mean ± SD, 2.4% ± 1.4%) and fascia thickness (5.0 ± 1.3 mm) values in the affected feet when compared with the unaffected feet in the patient group (VI, 1.4% ± 0.5%; fascia thickness, 3.3 ± 0.7 mm) and with the dominant side of the controls (VI, 1.6% ± 0.4%; fascia thickness, 2.9 ± 0.6 mm). After accounting for age, gender, body mass index, and duration of symptoms, the VI explained 13% and 33% of the variance in pain scores measured with a visual analog scale and the pain subscale of the Foot Function Index, respectively; the VI and fascia thickness explained 42% of the variance in the Foot Function Index. Individuals with unilateral chronic plantar fasciitis demonstrated significantly greater vascularity and thickened fascia on the affected side compared to the unaffected side and also to healthy controls. Fascia vascularity was associated independently with self-perceived pain, and both fascia vascularity and thickness were associated with foot dysfunction in patients with chronic plantar fasciitis. Public trials registry: Current Controlled Trials, ISRCTN49594569.


Clinical features of neuroendocrine prostate cancer.
Neuroendocrine prostate cancer (NEPC) is an aggressive variant of prostate cancer that may arise de novo or in patients previously treated with hormonal therapies for prostate adenocarcinoma as a mechanism of resistance. Despite being important to recognise, the clinical features of NEPC are poorly defined and could help guide when to perform a biopsy to look for NEPC histologic transformation. We reviewed baseline, treatment and outcome data of 87 patients with metastatic prostate cancer and tumour biopsy confirming NEPC histology. Forty-seven (54.0%) NEPC cases presented de novo, and 40 (46.0%) were therapy-related (t-NEPC). Thirty-six (41.4%) were classified as pure small-cell carcinoma, and 51 (58.6%) demonstrated mixed features with both small-cell carcinoma and adenocarcinoma present. Genomic data were available for 47 patients. The median age at time of NEPC was 68.1 years, median prostate-specific antigen (PSA) was 1.20 ng/ml (0.14 ng/mL small-cell carcinoma, 1.55 ng/mL mixed carcinoma) and sites of metastases included bone (72.6%), lymph node (47.0%), and viscera (65.5%). Median time from adenocarcinoma to t-NEPC diagnosis was 39.7 months (range, 24.5-93.8) with a median of two lines of prior systemic therapy. Platinum chemotherapy was used to treat 57.5% of patients, with a median progression-free survival of 3.9 months. Small-cell carcinoma was associated with worse overall survival (OS) than mixed histology (8.9 months from NEPC diagnosis versus 26.1 months, P < 0.001). Median OS of de novo NEPC was shorter than that of t-NEPC (16.8 months from prostate cancer diagnosis versus 53.5 months, P = 0.043). An average PSA rise per month of ≤0.7 ng/ml before t-NEPC; elevated lactate dehydrogenase levels, RB1 and TP53 loss and liver metastases were poor prognostic features. We describe the clinical features of a cohort of patients with NEPC. These characteristics may inform future diagnostic strategies.


An evaluation of My Strengths Training for Life™ for improving resilience and well-being of young people experiencing homelessness.
This community-based study investigated whether (1) a novel sport psychology informed positive youth development program, My Strengths Training for Life™, improved resilience and well-being and (2) young people differed in outcomes according to demographics (gender, ethnicity, social inclusion, and learning difficulty). A total of 246 young people (M age = 19.74, SD = 2.31) living in a large housing service completed questionnaires on demographics, mental skills, and pre and postprogram resilience and well-being. Baseline differences in resilience and well-being existed for ethnicity and learning difficulty status but did not influence MST4Life™ outcomes. There was a significant improvement in resilience and well-being over time, which was associated with mental skills development. Implications apply for policy, program commissioners, and research: (1) novel sport psychology interventions can improve the well-being of disadvantaged youth, and (2) demographics at baseline should be considered in intervention planning and evaluation with this population.


A chromosomal analysis of four species of Chilean Chrysomelinae (Coleoptera, Chrysomelidae).
Four species of Chilean leaf beetles in the subfamily Chrysomelinae have been cytogenetically analyzed, Blaptea elguetai Petitpierre, 2011, Henicotherus porteri Bréthes, 1929 and Jolivetia obscura (Philippi, 1864) show 2n = 28 chromosomes and a 13 + Xyp male meioformula, and Pataya nitida (Philippi, 1864) has the highest number of 2n = 38 chromosomes. The karyotype of Henicotherus porteri is made of mostly small meta/submetacentric chromosomes, and that of Jolivetia obscura displays striking procentric blocks of heterochromatin at pachytene autosomic bivalents using conventional staining. These findings are discussed in relation to previous cytogenetic data and current taxonomy of the subfamily.


Identification of isoflavone metabolites dihydrodaidzein, dihydrogenistein, 6'-OH-O-dma, and cis-4-OH-equol in human urine by gas chromatography-mass spectroscopy using authentic reference compounds.
The metabolic products of daidzein and genistein, the principal isoflavones of soy, were examined. Six volunteers included soy into their normal diet for a 2-week period and urine samples were analyzed before and after soy consumption. Isolation and characterization of the urinary metabolites were carried out with absorption chromatography on Sephadex LH-20 and gas chromatography-electron ionization mass spectrometry (GC-EIMS). The structures of the isoflavones isolated were confirmed by using authentic reference compounds. Dihydrogenistein, 6'-OH-O-desmethylangolensin, and cis-4-OH-equol were identified, in addition to known isoflavonoids daidzein, genistein, glycitein, and the known metabolites equol, O-desmethylangolensin, and dihydrodaidzein, by comparing the retention times and the spectra of the urinary compounds with those of the synthesized reference standards. The mammalian lignans enterolactone and enterodiol were also identified. Derivatization of the isoflavones for GC-MS was examined by comparing two silylating reagents, N, O-bis-(trimethylsilyl)-trifluoroacetamide (BSTFA) and pyridine:hexamethyldisilazan:trimethylchlorosilane (QSM), both used for the derivatization of these compounds. The silylation experiments revealed significant differences in the compositions of the derivatization products. Some corrections were made concerning the earlier published data of dihydrogenistein and 6'-OH-O-dma.


Spatio-temporal dynamics of yeast mitochondrial biogenesis: transcriptional and post-transcriptional mRNA oscillatory modules.
Examples of metabolic rhythms have recently emerged from studies of budding yeast. High density microarray analyses have produced a remarkably detailed picture of cycling gene expression that could be clustered according to metabolic functions. We developed a model-based approach for the decomposition of expression to analyze these data and to identify functional modules which, expressed sequentially and periodically, contribute to the complex and intricate mitochondrial architecture. This approach revealed that mitochondrial spatio-temporal modules are expressed during periodic spikes and specific cellular localizations, which cover the entire oscillatory period. For instance, assembly factors (32 genes) and translation regulators (47 genes) are expressed earlier than the components of the amino-acid synthesis pathways (31 genes). In addition, we could correlate the expression modules identified with particular post-transcriptional properties. Thus, mRNAs of modules expressed "early" are mostly translated in the vicinity of mitochondria under the control of the Puf3p mRNA-binding protein. This last spatio-temporal module concerns mostly mRNAs coding for basic elements of mitochondrial construction: assembly and regulatory factors. Prediction that unknown genes from this module code for important elements of mitochondrial biogenesis is supported by experimental evidence. More generally, these observations underscore the importance of post-transcriptional processes in mitochondrial biogenesis, highlighting close connections between nuclear transcription and cytoplasmic site-specific translation.


"Chase CRP", "Review patient": Improving the Quality of Weekend Medical Handover at a London Teaching Hospital.
Clinical handover has been identified as a "major preventable cause of harm" by the Royal College of Physicians (RCP). Whilst working at a London teaching hospital from August 2013, we noted substandard weekend handover of medical patients. The existing pro forma was filled incompletely by day doctors so it was difficult for weekend colleagues to identify unwell patients, with inherent safety implications. Furthermore, on-call medical staff noted that poor accessibility of vital information in patients' files was affecting acute clinical management. We audited the pro formas over a six week period (n=83) and the Friday ward round (WR) entries for medical inpatients over two weekends (n=84) against the RCP's handover guidance. The results showed poor documentation of several important details on the pro formas, for example, ceiling of care (4%) and past medical history (PMH) (23%). Problem lists were specified on 62% of the WR entries. We designed new handover pro formas and 'Friday WR sheets' to provide prompts for this information and used Medical Meetings and emails to explain the project's aims. Re-audit demonstrated significant improvement in all parameters; for instance, PMH increased to 52% on the pro formas. Only 10% of Friday WR entries used our sheet. However, when used, outcomes were much better, for example, problem list documentation increased to 100%. In conclusion, our interventions improved the provision of crucial information needed to prioritise and manage patients over the weekend. Future work should further highlight the importance of safe handover to all doctors to induce a shift in culture and optimise patient care.


Iodinated contrast media and the role of renal replacement therapy.
Iodinated contrast media are among the most commonly used pharmacologic agents in medicine. Although generally highly safe, iodinated contrast media are associated with several adverse effects, most significantly the risk of acute kidney injury, particularly in patients with underlying renal dysfunction. By virtue of their pharmacokinetic characteristics, these contrast agents are efficiently cleared by hemodialysis and to a lesser extent, hemofiltration. This has led to research into the capacity for renal replacement therapies to prevent certain adverse effects of iodinated contrast. This review examines the molecular and pharmacokinetic characteristics of iodinated contrast media and critically analyzes data from past studies on the role of renal replacement therapy to prevent adverse effects of these diagnostic agents.


Sulphate metabolism among thermophiles and hyperthermophiles in natural aquatic systems.
Although controversial, the idea that hydrothermal systems may have been the site for prebiotic synthesis of organic molecules and origin of life is widely supported. For the nascent life to survive, it must have had some sort of metabolic mechanism for generating energy. However, little is known of the specific metabolic pathways utilized by the early life forms or the effect of high temperatures on their activity. Recent research on natural high temperature aquatic environments, though limited because of difficult field logistics and experimental problems, is revolutionizing our understanding of possible energy-generating redox pathways, such as sulphate reduction. An abridged review of research on thermophilic sulphate reduction is presented here. Because of a complex interplay between microbiological and geochemical entities involved, and the uncertainties that modern hydrothermal systems are proxy for biogeochemical conditions on early Earth, great caution is required for interpretation and extrapolation of data from these studies to primordial times. Furthermore, a general lack of integrated geological and microbiological studies towards a common understanding of origin and sustenance of life on Earth is starkly evident from this review.


Chastising a child--the legal position.
Chastisement of children can take many forms, some of which could be offences under law. If the child was afraid of a physical attack because the parent was shouting at them, this could constitute an assault. A parent or other adult could face a charge of battery for physically striking a child. Under English law, reasonable chastisement is a defence available to a parent who may face legal prosecution for the action they take against their child. This article reviews the nature of chastisement and the current legal situation using relevant cases and related legislation. At present, a parent is entitled under English law to chastise their child; this includes smacking them where doing so leaves no mark on the child.


Assessment of gefitinib- and CI-1040-mediated changes in epidermal growth factor receptor signaling in HuCCT-1 human cholangiocarcinoma by serial fine needle aspiration.
One specific limitation to the clinical development of targeted cancer therapeutics is the lack of well-validated pharmacodynamic markers. Such tools might conceivably provide a framework within which to better evaluate the selection of specific molecules as therapeutic targets. Nevertheless, the practical application of this hypothesis in clinical development remains elusive. In this study, we present a minimally invasive pharmacodynamic assay for monitoring therapy-mediated changes in the activity of target signaling pathways by using fine needle aspiration (FNA) samples and quantitative ELISA methods. To this end, we used the HuCCT-1 cholangiocarcinoma cell line treated with gefitinib (ZD1839, Iressa), a selective blocker of the epidermal growth factor receptor (EGFR), and CI-1040, a selective inhibitor of the mitogen extracellular regulated kinase [mitogen-activated protein/extracellular signal-regulated kinase (ERK) kinase 1/2]. HuCCT-1 cells were resistant to gefitinib and CI-1040 alone but susceptible to the combination of these drugs in vitro and in vivo. This effect was associated with a greater inhibition of ERK1/2 activation, a downstream mediator in the EGFR-mitogen-activated protein/ERK kinase pathway. Using this model, we sought to assess whether FNA-obtained tumor biopsies could be used to measure signaling pathway activation. Cellular extracts prepared from FNA samples yielded adequately cellular, high-quality samples to assess therapy-mediated changes in EGFR and ERK1/2 phosphorylation by Western blotting and quantitative ELISA assays. Treatment with gefitinib alone effectively inhibited EGFR activation but failed to block ERK1/2 phosphorylation and tumor growth. Blocking was achieved by the addition of CI-1040 to the treatment regimen. These results show that the combination of serial FNA sampling with highly sensitive quantitative ELISA assays permits assessment of therapy-mediated changes in signaling pathways, which correlate well with antitumor effects. This assay is simple to implement and broadly applicable to diverse tumor types in clinical studies with cancer patients and may be useful in the development of targeted anticancer agents.


Kruppel-like factor 4 regulates endothelial inflammation.
The vascular endothelium plays a critical role in vascular homeostasis. Inflammatory cytokines and non-laminar blood flow induce endothelial dysfunction and confer a pro-adhesive and pro-thrombotic phenotype. Therefore, identification of factors that mediate the effects of these stimuli on endothelial function is of considerable interest. Kruppel-like factor 4 expression has been documented in endothelial cells, but a function has not been described. In this communication we describe the expression in vitro and in vivo of Kruppel-like factor 4 in human and mouse endothelial cells. Furthermore, we demonstrate that endothelial Kruppel-like factor 4 is induced by pro-inflammatory stimuli and shear stress. Overexpression of Kruppel-like factor 4 induces expression of multiple anti-inflammatory and anti-thrombotic factors including endothelial nitric-oxide synthase and thrombomodulin, whereas knockdown of Kruppellike factor 4 leads to enhancement of tumor necrosis factor alpha-induced vascular cell adhesion molecule-1 and tissue factor expression. The functional importance of Kruppel-like factor 4 is verified by demonstrating that Kruppel-like factor 4 expression markedly decreases inflammatory cell adhesion to the endothelial surface and prolongs clotting time under inflammatory states. Kruppel-like factor 4 differentially regulates the promoter activity of pro- and anti-inflammatory genes in a manner consistent with its anti-inflammatory function. These data implicate Kruppel-like factor 4 as a novel regulator of endothelial activation in response to pro-inflammatory stimuli.


A 160-kilodalton epithelial cell surface glycoprotein recognized by plant lectins that inhibit the adherence of Actinomyces naeslundii.
The adherence of Actinomyces naeslundii to human epithelial (KB) cells is mediated by the interaction of a fimbrial lectin on this oral bacterium with epithelial cell receptors exposed by sialidase. The D-galactose- and N-acetyl-D-galactosamine-reactive plant lectins from peanut and from Bauhinia purpurea inhibit this interaction. This report describes the partial purification and characterization of a 160-kilodalton (kDa) cell surface glycoprotein which is the principal receptor for these lectins. Radioiodinated lectins detected a band of 160 kDa on sialidase-treated Western blots of epithelial cell extracts but did not detect bands on nontreated filters. However, wheat germ agglutinin was reactive with the 160-kDa band on filters that were not treated with sialidase, suggesting that this lectin recognizes the sialic acid residues of this molecule. The 160-kDa component was partially purified from n-octylglucoside extracts of the epithelial cells by wheat germ agglutinin affinity chromatography. This molecule was metabolically labeled with D-[14C]glucosamine and labeled at the cell surface by lactoperoxidase-catalyzed iodination or periodate oxidation followed by sodium borotritide reduction. Incubation of epithelial cells with sialidase before extraction resulted in the loss of the 160-kDa band and the appearance of a band at 200 kDa which was directly reactive with 125I-labeled peanut agglutinin. These results indicate that the 160-kDa glycoprotein on the surface of the epithelial cell serves as a receptor for the agglutinins from the peanut and B. purpurea and presumably the fimbrial lectin of actinomyces.


Sleep quality burden in chronic rhinosinusitis with nasal polyps and its modulation by dupilumab.
Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with a substantial burden on patients' quality of life and impaired sleep quality. The most common CRSwNP endotype is characterized by type 2 inflammation, with enhanced production of interleukin (IL)-4, IL-5, and IL-13. Dupilumab is a monoclonal antibody against IL-4 receptor-α, which inhibits both IL-4 and IL-13 signaling, and was recently approved for treatment of CRSwNP. We investigated the effect of dupilumab on the sleep quality of patients with CRSwNP in a real-life setting. Patients were evaluated at baseline and after 1 and 3 months of dupilumab treatment by means of the Epworth sleepiness scale (ESS), insomnia severity index (ISI), Pittsburgh sleep quality index (PSQI), and sinonasal outcome test 22 (SNOT-22) sleep domain. A total of 29 consecutive patients were enrolled, and their baseline sleep quality assessment were as follows: ESS of 7.9 (± 4.5); ISI of 13.1 (± 6.2); PSQI of 9.2 (± 3.7); and SNOT-22 sleep domain of 12.1 (± 4.2). Excessive daily sleepiness, insomnia, and globally impaired sleep quality were present in 24.1%, 79.3%, and 93.1% respectively. Treatment with dupilumab was associated with significant improvement in ESS, ISI, PSQI, and SNOT-22 sleep domain with concomitant reduction of the proportion of patients with insomnia and globally impaired sleep quality. CRSwNP was associated with a significant impact on global sleep quality, in particular, insomnia, and treatment with dupilumab induced a rapid improvement (after 1 single month of treatment) in all the sleep quality parameters, suggesting that sleep disturbances should be more carefully evaluated as an additional outcome in these patients.


Cryptosporidiosis in young artiodactyls.
Cryptosporidium was found in the intestinal tract of 10 blackbuck, 2 scimitar-horned oryx, 2 fringe-eared oryx, 2 addax, and 1 sable antelope that had diarrhea. Cryptosporidia were most numerous in the small intestine, but also were found in the cecum, spiral colon, and colon. The small intestine had minimal inflammation in association with the cryptosporidia. Salmonella typhimurium was isolated from 10 of the 51 animals evaluated, with extensive inflammation of the cecum, spiral colon, and colon observed in these animals.


A truncated aminoacyl-tRNA synthetase modifies RNA.
Aminoacyl-tRNA synthetases are modular enzymes composed of a central active site domain to which additional functional domains were appended in the course of evolution. Analysis of bacterial genome sequences revealed the presence of many shorter aminoacyl-tRNA synthetase paralogs. Here we report the characterization of a well conserved glutamyl-tRNA synthetase (GluRS) paralog (YadB in Escherichia coli) that is present in the genomes of >40 species of proteobacteria, cyanobacteria, and actinobacteria. The E. coli yadB gene encodes a truncated GluRS that lacks the C-terminal third of the protein and, consequently, the anticodon binding domain. Generation of a yadB disruption showed the gene to be dispensable for E. coli growth in rich and minimal media. Unlike GluRS, the YadB protein was able to activate glutamate in presence of ATP in a tRNA-independent fashion and to transfer glutamate onto tRNA(Asp). Neither tRNA(Glu) nor tRNA(Gln) were substrates. In contrast to canonical aminoacyl-tRNA, glutamate was not esterified to the 3'-terminal adenosine of tRNA(Asp). Instead, it was attached to the 2-amino-5-(4,5-dihydroxy-2-cyclopenten-1-yl) moiety of queuosine, the modified nucleoside occupying the first anticodon position of tRNA(Asp). Glutamyl-queuosine, like canonical Glu-tRNA, was hydrolyzed by mild alkaline treatment. Analysis of tRNA isolated under acidic conditions showed that this novel modification is present in normal E. coli tRNA; presumably it previously escaped detection as the standard conditions of tRNA isolation include an alkaline deacylation step that also causes hydrolysis of glutamyl-queuosine. Thus, this aminoacyl-tRNA synthetase fragment contributes to standard nucleotide modification of tRNA.


Randomized trial of the effect of contact lens wear on self-perception in children.
To determine whether contact lens wear affects children's self-perceptions. The Adolescent and Child Health Initiative to Encourage Vision Empowerment Study was a randomized, single-masked trial conducted at five clinical centers in the United States. Subjects were 8- to 11-year-old myopic children randomly assigned to wear spectacles (n = 237) or soft contact lenses (n = 247) for 3 years. The primary endpoint was the Self-Perception Profile for Children Global Self-Worth scale. Secondary outcomes included the Physical Appearance, Athletic Competence, Scholastic Competence, Behavioral Conduct, and Social Acceptance Self-Perception Profile for Children scales. Global self-worth was not affected by contact lens wear [analysis of variance (ANOVA), difference = 0.06; 95% CI, -0.004 to 0.117]. Physical appearance (ANOVA, difference = 0.15; 95% CI, 0.07 to 0.22), athletic competence (ANOVA, difference = 0.08; 95% CI, 0.01 to 0.15), and social acceptance (ANOVA, difference = 0.10; 95% CI, 0.03 to 0.17) were all greater for contact lens wearers. Although contact lens wear does not affect global self-perceptions of 8- to 11-year-old myopic children their physical appearance, athletic competence, and social acceptance self-perceptions are likely to improve with contact lens wear. Eye care practitioners should consider the social and visual benefits of contact lens wear when choosing the most appropriate vision correction modality for children as young as 8 years of age.