Evaluation of the effectiveness of manual chest physiotherapy techniques on quality of life at six months post exacerbation of COPD (MATREX): a randomised controlled equivalence trial.
Manual chest physiotherapy (MCP) techniques involving chest percussion, vibration, and shaking have long been used in the treatment of respiratory conditions. However, methodological limitations in existing research have led to a state of clinical equipoise with respect to this treatment. Thus, for patients hospitalised with an exacerbation of Chronic Obstructive Pulmonary Disease (COPD), clinical preference tends to dictate whether MCP is given to assist with sputum clearance. We standardised the delivery of MCP and assessed its effectiveness on disease-specific quality of life. In this randomised, controlled trial powered for equivalence, 526 patients hospitalised with acute COPD exacerbation were enrolled from four centres in the UK. Patients were allocated to receive MCP plus advice on airway clearance or advice on chest clearance alone. The primary outcome was a COPD specific quality of life measure, the Saint Georges Respiratory Questionnaire (SGRQ) at six months post randomisation. Analyses were by intention to treat (ITT). This study was registered, ISRCTN13825248. All patients were included in the analyses, of which 372 (71%) provided evaluable data for the primary outcome. An effect size of 0·3 standard deviations in SGRQ score was specified as the threshold for superiority. The ITT analyses showed no significant difference in SGRQ for patients who did, or did not receive MCP (95% CI -0·14 to 0·19). These data do not lend support to the routine use of MCP in the management of acute exacerbation of COPD. However, this does not mean that MCP is of no therapeutic value to COPD patients in specific circumstances.


A mathematical model gives insights into nutritional and genetic aspects of folate-mediated one-carbon metabolism.
Impaired folate-mediated 1-carbon metabolism has been linked to multiple disease outcomes. A better understanding of the nutritional and genetic influences on this complex biochemical pathway is needed to comprehend their impact on human health. To this end, we created a mathematical model of folate-mediated 1-carbon metabolism. The model uses published data on folate enzyme kinetics and regulatory mechanisms to simulate the impact of genetic and nutritional variation on critical aspects of the pathway. We found that the model predictions match experimental data, while providing novel insights into pathway kinetics. Our primary observations were as follows: 1) the inverse association between folate and homocysteine is strongest at very low folate concentrations, but there is no association at high folate concentrations; 2) the DNA methylation reaction rate is relatively insensitive to changes in folate pool size; and 3) as folate concentrations become very high, enzyme velocities decrease. With regard to polymorphisms in 5,10-methylenetetrahydrofolate reductase (MTHFR), the modeling predicts that decrease MTHFR activity reduces concentrations of S-adenosylmethionine and 5-methyltetrahydrofolate, as well as DNA methylation, while modestly increasing S-adenosylhomocysteine and homocysteine concentrations and thymidine or purine synthesis. Decreased folate together with a simulated vitamin B-12 deficiency results in decreases in DNA methylation and purine and thymidine synthesis. Decreased MTHFR activity superimposed on the B-12 deficiency appears to reverse the declines in purine and thymidine synthesis. These mathematical simulations of folate-mediated 1-carbon metabolism provide a cost-efficient approach to in silico experimentation that can complement and help guide laboratory studies.


Detecting hybridization between wild species and their domesticated relatives.
The widespread occurrence of free-ranging domestic or feral carnivores (dogs, cats) or ungulates (pigs, goats), and massive releases of captive-reproduced game stocks (galliforms, waterfowl) is raising fear that introgressive hybridization with wild populations might disrupt local adaptations, leading to population decline and loss of biodiversity. Detecting introgression through hybridization is problematic if the parental populations cannot be sampled (unlike in classical stable hybrid zones), or if hybridization is sporadic. However, the use of hypervariable DNA markers (microsatellites) and new statistical methods (Bayesian models), have dramatically improved the assessment of cryptic population structure, admixture analyses and individual assignment testing. In this paper, I summarize results of projects aimed to identify occurrence and extent of introgressive hybridization in European populations of wolves (Canis lupus), wildcats (Felis silvestris), rock partridges and red-legged partridges (Alectoris graeca and Alectoris rufa), using genetic methods. Results indicate that introgressive hybridization can be locally pervasive, and that conservation plans should be implemented to preserve the integrity of the gene pools of wild populations. Population genetic methods can be fruitfully used to identify introgressed individuals and hybridizing populations, providing data which allow evaluating risks of outbreeding depression. The diffusion in the wild of invasive feral animals, and massive restocking with captive-reproduced game species, should be carefully controlled to avoid loss of genetic diversity and disruption of local adaptations.


The role of hyaluronic acid, chitosan, and calcium sulfate and their combined effect on early bony consolidation in distraction osteogenesis of a canine model.
The purpose of this project was to study the effect of hyaluronic acid, calcium sulfate, and chitosan on early bony consolidation in distraction osteogenesis of a canine model. Sixteen dogs were used for this study. The lateral surface of the mandibular body was exposed in the subperiosteal plane, and the vertical osteotomy on the mandibular body was extended downward. An external distraction device was applied to the mandibular body, and the mandibular distraction was started 5 days after the operation at a rate of 1 mm/d up to a 10-mm distraction. The experimental group was then divided into a control group, chitosan group, hyaluronic acid group, calcium sulfate combined with hyaluronic acid group, and calcium sulfate combined with chitosan group, depending on the type of implantation material in the distracted area. After completing the distraction, implantation material was injected into the distracted area, although no material was implanted into the distracted area of the control group. After implanting the materials, the distraction device was left in place for 6 weeks to allow for bony consolidation. Four dogs were allocated to each group. Two dogs in each group (total of 8 dogs) were killed 3 weeks after implantation of the material, and the other 8 dogs were killed after 6 weeks. New bone was generated in the distracted zone of all groups. In the calcium sulfate combined with chitosan group and calcium sulfate combined with hyaluronic acid group, the formation of active woven bone was observed throughout the distracted zone. Moreover, the new bone seemed to be nearly normal cortical bone at 6 weeks after implantation. In the chitosan group and hyaluronic acid group, the development of new bone was observed in the distracted zone at 6 weeks. The amount was less than that in the calcium sulfate combined with hyaluronic acid group and calcium sulfate combined with chitosan group. These findings suggest that calcium sulfate and its combined materials seem to be quite effective in early bony consolidation in distraction osteogenesis.


Electro-optical switching by liquid-crystal controlled metasurfaces.
We study the optical response of a metamaterial surface created by a lattice of split-ring resonators covered with a nematic liquid crystal and demonstrate millisecond timescale switching between electric and magnetic resonances of the metasurface. This is achieved due to a high sensitivity of liquid-crystal molecular reorientation to the symmetry of the metasurface as well as to the presence of a bias electric field. Our experiments are complemented by numerical simulations of the liquid-crystal reorientation.


Fasting might not be necessary before lipid screening: a nationally representative cross-sectional study.
There are barriers to fasting lipid screening for at-risk children. Results of studies in adults have suggested that lipid testing might be reliably performed without fasting. To examine population-level differences in pediatric lipid values based on length of fast before testing. We used the National Health and Nutrition Examination Survey (1999-2008) to examine total cholesterol (TC), HDL (high-density lipoprotein), LDL (low-density lipoprotein), and triglyceride cholesterol components on the basis of the period of fasting. Young children fasted for varying times before being tested, and children older than 12 years were asked to fast; however, adherence was variable. We used ordinary least-squares regression to test for differences in lipid values that were based on fasting times, controlling for weight status, age, race, ethnicity, and gender. TC, HDL, LDL, or triglyceride values were available for 12 744 children. Forty-eight percent of the TC and HDL samples and 80% of the LDL and triglyceride samples were collected from children who had fasted ≥ 8 hours. Fasting had a small positive effect for TC, HDL, and LDL, resulting in a mean value for the sample that was 2 to 5 mg/dL higher with a 12-hour fast compared with a no-fast sample. Fasting time had a negative effect on triglycerides (β = -0.859; P = .02), which resulted in values in the fasting group that were 7 mg/dL lower. Comparison of cholesterol screening results for a nonfasting group of children compared with results for a similar fasting group resulted in small differences that are likely not clinically important. Physicians might be able to decrease the burden of childhood cholesterol screening by not requiring prescreening fasting for these components.


Assessing uncertainty in model parameters based on sparse and noisy experimental data.
To perform parametric identification of mathematical models of biological events, experimental data are rare to be sufficient to estimate target behaviors produced by complex non-linear systems. We performed parameter fitting to a cell cycle model with experimental data as an in silico experiment. We calibrated model parameters with the generalized least squares method with randomized initial values and checked local and global sensitivity of the model. Sensitivity analyses showed that parameter optimization induced less sensitivity except for those related to the metabolism of the transcription factors c-Myc and E2F, which are required to overcome a restriction point (R-point). We performed bifurcation analyses with the optimized parameters and found the bimodality was lost. This result suggests that accumulation of c-Myc and E2F induced dysfunction of R-point. We performed a second parameter optimization based on the results of sensitivity analyses and incorporating additional derived from recent in vivo data. This optimization returned the bimodal characteristics of the model with a narrower range of hysteresis than the original. This result suggests that the optimized model can more easily go through R-point and come back to the gap phase after once having overcome it. Two parameter space analyses showed metabolism of c-Myc is transformed as it can allow cell bimodal behavior with weak stimuli of growth factors. This result is compatible with the character of the cell line used in our experiments. At the same time, Rb, an inhibitor of E2F, can allow cell bimodal behavior with only a limited range of stimuli when it is activated, but with a wider range of stimuli when it is inactive. These results provide two insights; biologically, the two transcription factors play an essential role in malignant cells to overcome R-point with weaker growth factor stimuli, and theoretically, sparse time-course data can be used to change a model to a biologically expected state.


ABO blood group antigens on human plasma von Willebrand factor after ABO-mismatched bone marrow transplantation.
von Willebrand factor (vWF) is synthesized exclusively by endothelial cells and megakaryocytes, and stored in the intracellular granules or constitutively secreted into plasma. ABO blood group antigens are covalently associated with asparagine-linked sugar chains of plasma vWF. The effect of ABO-mismatched bone marrow transplantation (BMT) or blood stem cell transplantation (BSCT) on the expression of ABO blood group antigens on the vWF was examined to obtain information on the origin of these antigens. In ABO-mismatched (HLA-matched) groups, 8 cases of BMT and 4 cases of BSCT were examined. In all cases, the ABO blood groups on red blood cells were gradually converted to the donor's type within 80 to 90 days after the transplantation. The blood group antigens on the vWF were consistent with the recipient's blood group for the period monitored by enzyme-linked immunosorbent assay (ELISA). When vWF was isolated from normal platelets and examined for the blood group antigens using ELISA or immunoblotting, it showed few antigens. However, vWF extracted from veins expressed blood group antigens. These findings indicate that platelet (megakaryocyte)-derived vWF does not contain blood group antigens and that these antigens may be specifically associated with vWF synthesized in endothelial cells and secreted into plasma. Furthermore, it is possible that the persistence of the recipient's blood group antigens on plasma glycoproteins such as vWF, independent of the donor-derived erythrocytes, after ABO-mismatched stem cell transplantation, may influence the immunological system in the production of anti-blood group antibodies resulting in the establishment of immunological tolerance in the recipient plasma.


Effect of Axillary Dissection vs No Axillary Dissection on 10-Year Overall Survival Among Women With Invasive Breast Cancer and Sentinel Node Metastasis: The ACOSOG Z0011 (Alliance) Randomized Clinical Trial.
The results of the American College of Surgeons Oncology Group Z0011 (ACOSOG Z0011) trial were first reported in 2005 with a median follow-up of 6.3 years. Longer follow-up was necessary because the majority of the patients had estrogen receptor-positive tumors that may recur later in the disease course (the ACOSOG is now part of the Alliance for Clinical Trials in Oncology). To determine whether the 10-year overall survival of patients with sentinel lymph node metastases treated with breast-conserving therapy and sentinel lymph node dissection (SLND) alone without axillary lymph node dissection (ALND) is noninferior to that of women treated with axillary dissection. The ACOSOG Z0011 phase 3 randomized clinical trial enrolled patients from May 1999 to December 2004 at 115 sites (both academic and community medical centers). The last date of follow-up was September 29, 2015, in the ACOSOG Z0011 (Alliance) trial. Eligible patients were women with clinical T1 or T2 invasive breast cancer, no palpable axillary adenopathy, and 1 or 2 sentinel lymph nodes containing metastases. All patients had planned lumpectomy, planned tangential whole-breast irradiation, and adjuvant systemic therapy. Third-field radiation was prohibited. The primary outcome was overall survival with a noninferiority hazard ratio (HR) margin of 1.3. The secondary outcome was disease-free survival. Among 891 women who were randomized (median age, 55 years), 856 (96%) completed the trial (446 in the SLND alone group and 445 in the ALND group). At a median follow-up of 9.3 years (interquartile range, 6.93-10.34 years), the 10-year overall survival was 86.3% in the SLND alone group and 83.6% in the ALND group (HR, 0.85 [1-sided 95% CI, 0-1.16]; noninferiority P = .02). The 10-year disease-free survival was 80.2% in the SLND alone group and 78.2% in the ALND group (HR, 0.85 [95% CI, 0.62-1.17]; P = .32). Between year 5 and year 10, 1 regional recurrence was seen in the SLND alone group vs none in the ALND group. Ten-year regional recurrence did not differ significantly between the 2 groups. Among women with T1 or T2 invasive primary breast cancer, no palpable axillary adenopathy, and 1 or 2 sentinel lymph nodes containing metastases, 10-year overall survival for patients treated with sentinel lymph node dissection alone was noninferior to overall survival for those treated with axillary lymph node dissection. These findings do not support routine use of axillary lymph node dissection in this patient population based on 10-year outcomes. clinicaltrials.gov Identifier: NCT00003855.


Partnering with governments and other institutions: driving change in diabetes care.
The human and material cost of type 2 diabetes is a cause of increasing concern for health professionals, representative organisations and governments worldwide. The scale of morbidity and mortality has led the United Nations to issue a resolution on diabetes, calling for national policies for prevention, treatment and care. There is clearly an urgent need for a concerted response from all interested parties at the community, national and international level to work towards the goals of the resolution and create effective, sustainable treatment models, care systems and prevention strategies. Action requires both a 'bottom-up' approach of public awareness campaigns and pressure from healthcare professionals, coupled with a 'top-down' drive for change, via partnerships with governments, third sector (non-governmental) organisations and other institutions. In this review, we examine how existing collaborative initiatives serve as examples for those seeking to implement change in health policy and practice in the quest to alleviate the health and economic burden of diabetes. Efforts are underway to provide continuous and comprehensive care models for those who already have type 2 diabetes; in some cases, national plans extend to prevention strategies in attempts to improve overall public health. In the spirit of partnership, collaborations with governments that incorporate sustainability, long-term goals and a holistic approach continue to be a driving force for change. It is now critical to maintain this momentum and use the growing body of compelling evidence to educate, inform and deliver a long-term, lasting impact on patient and public health worldwide.


Hazard identification of pyrogenic synthetic amorphous silica (NM-203) after sub-chronic oral exposure in rat: A multitarget approach.
Food additive E551 consists of synthetic amorphous silica (SAS), comprising agglomerates and aggregates of primary particles in the nanorange (<100 nm), which potential nanospecific risks for humans associated to dietary exposure are not yet completely assessed. In NANoREG project, aim of the study was to identify potential hazards of pyrogenic SAS nanomaterial NM-203 by a 90-day oral toxicity study (OECD test guideline 408). Adult Sprague-Dawley rats of both sexes were orally treated with 0, 2, 5, 10, 20 and 50 mg SAS/kg bw per day; dose levels were selected to be as close as possible to E551 dietary exposure. Several endpoints were investigated, the whole integrative study is presented here along with the results of dispersion characterization, tissue distribution, general toxicity, blood/serum biomarkers, histopathological and immunotoxicity endpoints. No mortality, general toxicity and limited deposition in target tissues were observed. NM-203 affected liver and spleen in both sexes. Proposed NOAEL 5 mg/kg bw per day in male rats for enlarged sinusoids in liver. In female rats, TSH and creatinine levels were affected, proposed LOAEL 2 mg/kg bw per day. Overall, these data provide new insight for a comprehensive risk assessment of SAS exposure by the oral route.


Randomised, double-blind, placebo-controlled study of olanzapine in patients with bipolar I depression.
Atypical antipsychotics are widely used in bipolar mania. However, the efficacy of atypical antipsychotics in bipolar depression has not been comprehensively explored. To evaluate olanzapine monotherapy in patients with bipolar depression. Patients with bipolar depression received olanzapine (5-20 mg/day, n = 343) or placebo (n = 171) for 6 weeks. The primary outcome was change from baseline to end-point in Montgomery-Åsberg Depression Rating Scale (MADRS) total score. Secondary outcomes included: Clinical Global Impression - Bipolar Version (CGI-BP) scale, 17-item Hamilton Rating Scale for Depression (HRSD-17) and Young Mania Rating Scale (YMRS) scores, and the rate of response (≥50% reduction in MADRS at end-point), recovery (MADRS ≤12 for ≥4 weeks plus treatment completion) and remission (MADRS ≤8). The trial was registered with ClinicalTrials.gov (NCT00510146). Olanzapine demonstrated: significantly greater (P<0.04) improvements on MADRS (least-squares mean change -13.82 v. -11.67), HRSD-17 and YMRS total scores and all CGI-BP subscale scores v. placebo; significantly (P≤0.05) more response and remission, but not recovery; significantly (P<0.01) greater mean increases in weight, fasting cholesterol and triglycerides; and significantly more (P<0.001) patients gained ≥7% body weight. Olanzapine monotherapy appears to be efficacious in bipolar depression. Additional long-term studies are warranted to confirm these results. Safety findings were consistent with the known safety profile of olanzapine.


Rabbits immunized with a peptide encoded for by the 230-kD bullous pemphigoid antigen cDNA develop an enhanced inflammatory response to UVB irradiation: a potential animal model for bullous pemphigoid.
Previous attempts to develop an animal model of bullous pemphigoid (BP) have failed to result in inflammatory disease in the skin. P1-2 is an 18-amino acid peptide encoded for by the 230-kD BP antigen cDNA that has been shown to contain an epitope recognized by circulating antibodies from patients with BP. The purpose of this study was to determine if ultraviolet B irradiation of rabbits after immunization with the P1-2 peptide would result in an enhanced inflammatory response in the skin to that injury. Three rabbits were immunized with either P1-2 or a control peptide. All rabbits immunized with P1-2, and none of the control rabbits, developed antibodies against P1-2 that bound in vitro to both human and rabbit skin in a linear pattern at the basement membrane zone. Immunized rabbits were irradiated on the flank with ultraviolet light. Rabbits immunized with P1-2 developed an enhanced inflammatory reaction to ultraviolet B irradiation leading to epidermal necrosis and sloughing of some sites in 6-9 d. Control rabbits showed only mild erythema without sloughing, which healed in 4-6 d. Histology in the P1-2 immunized rabbits at 24 h revealed an inflammatory infiltrate of neutrophils at the dermal-epidermal junction, whereas control rabbits showed only mild edema and a sparse inflammatory infiltrate. All the rabbits immunized with P1-2 had linear deposits of immunoglobulin G and C3 at the basement membrane zone of healed skin compared to none of the controls. These findings demonstrate that antibodies against a synthetic peptide encoded by the BP antigen 1 sequence can lead to an enhanced inflammatory response after epithelial injury in rabbit skin.


Executive dysfunction and treatment response in late-life depression.
Executive dysfunction in geriatric depression has been shown to predict poor response to antidepressant medication. The purpose of this review is to clarify which aspects of executive functioning predict poor antidepressant treatment response. Literature review. From our review, the aspects of executive functioning that appear to be associated with antidepressant response rates are verbal fluency and response inhibition. There is some indication that the semantic strategy component may account for the effects of verbal fluency, although evidence comes from one study and needs replication. Processing speed has been proposed as a substrate that may underlie the effects of executive dysfunction on treatment response. Although processing speed does not appear to account for the relationship between response inhibition and treatment outcome, this issue has yet to be assessed with respect to verbal fluency. Verbal fluency and response inhibition are specific aspects of executive dysfunction that appear to impact antidepressant response rates. Disruption of the frontostriatal limbic circuit (particularly the anterior cingulate and dorsolateral prefrontal cortex) may explain the relation between these two mechanisms.


Unravelling the complex story of intergenomic recombination in ABB allotriploid bananas.
Bananas (Musa spp.) are a major staple food for hundreds of millions of people in developing countries. The cultivated varieties are seedless and parthenocarpic clones of which the ancestral origin remains to be clarified. The most important cultivars are triploids with an AAA, AAB or ABB genome constitution, with A and B genomes provided by M. acuminata and M. balbisiana, respectively. Previous studies suggested that inter-genome recombinations were relatively common in banana cultivars and that triploids were more likely to have passed through an intermediate hybrid. In this study, we investigated the chromosome structure within the ABB group, composed of starchy cooking bananas that play an important role in food security. Using SNP markers called from RADSeq data, we studied the chromosome structure of 36 ABB genotypes spanning defined taxonomic subgroups. To complement our understanding, we searched for similar events within nine AB hybrid genotypes. Recurrent homologous exchanges (HEs), i.e. chromatin exchanges between A and B subgenomes, were unravelled with at least nine founding events (HE patterns) at the origin of ABB bananas prior to clonal diversification. Two independent founding events were found for Pisang Awak genotypes. Two HE patterns, corresponding to genotypes Pelipita and Klue Teparod, show an over-representation of B genome contribution. Three HE patterns mainly found in Indian accessions shared some recombined regions and two additional patterns did not correspond to any known subgroups. The discovery of the nine founding events allowed an investigation of the possible routes that led to the creation of the different subgroups, which resulted in new hypotheses. Based on our observations, we suggest different routes that gave rise to the current diversity in the ABB cultivars, routes involving primary AB hybrids, routes leading to shared HEs and routes leading to a B excess ratio. Genetic fluxes took place between M. acuminata and M. balbisiana, particularly in India, where these unbalanced AB hybrids and ABB allotriploids originated, and where cultivated M. balbisiana are abundant. The result of this study clarifies the classification of ABB cultivars, possibly leading to the revision of the classification of this subgroup.


Deletion and mutation analyses of the P16/MTS-1 tumor suppressor gene in human ductal pancreatic cancer reveals a higher frequency of abnormalities in tumor-derived cell lines than in primary ductal adenocarcinomas.
The putative tumor suppressor gene p16/CDKN2 encodes a specific inhibitor of cyclin D-cyclin-dependent kinase 4 complexes important in cell-cycle regulation and has been found to be deleted or mutated in a variety of human cancers. Thirty microdissected primary human ductal pancreatic carcinomas from patients not subject to radiotherapy or chemotherapy prior to surgical resection of their carcinomas and 18 human pancreatic carcinoma cell lines were analyzed by single-strand conformation polymorphism (SSCP) and DNA sequence analyses and PCR-based deletion analyses for mutations and homozygous deletions of the p16/CDKN2 gene, respectively. Homozygous deletions of the gene were found in five cell lines, and nonpolymorphic SSCP and DNA sequence alterations were found within exon 1 in four cell lines and exon 2 in three lines, for an overall frequency of deletions and mutations of 66%. In contrast, homozygous deletions of p16/CDKN2 were observed in three primary pancreatic carcinomas, and five primary tumors revealed SSCP and/or sequence abnormalities in exon 1 (one case) and exon 2 (four cases), a mutation and deletion frequency of 27%. Immunoblotting analyses confirmed the absence of p16/MTS-1 expression in actively proliferating cell lines with a homozygous deletion of the gene and low-to-moderate levels of p16/MTS-1 expression in cell lines possessing a normal RB-1 gene or protein. These findings suggest that, although p16/CDKN2 may play a role in the pathobiology of pancreatic cancer, inactivation of this putative tumor suppressor gene occurs more frequently in cell lines than in primary ductal pancreatic carcinomas.


Carotenoid dietary intakes and plasma concentrations are associated with heel bone ultrasound attenuation and osteoporotic fracture risk in the European Prospective Investigation into Cancer and Nutrition (EPIC)-Norfolk cohort.
Carotenoids are found in abundance in fruit and vegetables, and may be involved in the positive association of these foods with bone health. This study aimed to explore the associations of dietary carotenoid intakes and plasma concentrations with bone density status and osteoporotic fracture risk in a European population. Cross-sectional analyses (n 14 803) of bone density status, using calcaneal broadband ultrasound attenuation (BUA) and longitudinal analyses (n 25 439) of fracture cases were conducted on data from the prospective European Prospective Investigation into Cancer and Nutrition-Norfolk cohort of middle-aged and older men and women. Health and lifestyle questionnaires were completed, and dietary nutrient intakes were derived from 7-d food diaries. Multiple regression demonstrated significant positive trends in BUA for women across quintiles of dietary α-carotene intake (P=0·029), β-carotene intake (P=0·003), β-cryptoxanthin intake (P=0·031), combined lutein and zeaxanthin intake (P=0·010) and lycopene intake (P=0·005). No significant trends across plasma carotenoid concentration quintiles were apparent (n 4570). The Prentice-weighted Cox regression showed no trends in fracture risk across dietary carotenoid intake quintiles (mean follow-up time 12·5 years), except for a lower risk for wrist fracture in women with higher lutein and zeaxanthin intake (P=0·022); nevertheless, inter-quintile differences in fracture risk were found for both sexes. Analysis of plasma carotenoid data (mean follow-up time 11·9 years) showed lower hip fracture risk in men across higher plasma α-carotene (P=0·026) and β-carotene (P=0·027) quintiles. This study provides novel evidence that dietary carotenoid intake is relevant to bone health in men and women, demonstrating that associations with bone density status and fracture risk exist for dietary intake of specific carotenoids and their plasma concentrations.


Diagnosis of Legionella lung abscess by percutaneous needle aspiration.
Pneumonia is a life-threatening infection of immunocompromised patients, and accurate identification of the causative agent is essential for early initiation of appropriate antibiotic therapy. Percutaneous needle aspiration was used to identify Legionella pneumonia after sputum and bronchoscopy specimens had failed to provide a diagnosis in a 13-year-old leukemic patient with a lung abscess. We believe this is the first report of a child with this disease diagnosed in this way.


Temporal Expectation Improves Recognition Memory for Spatially Attended Objects.
Recent evidence suggests that temporal expectation is beneficial to memory formation. Rhythmic presentation of stimuli during encoding enhances subsequent recognition and is associated with distinct neural activity compared with when stimuli are presented in an arrhythmic manner. However, no prior study has examined how temporal expectation interacts with another important form of facilitation-spatial attention-to affect memory. This study systematically manipulated temporal expectation and spatial attention during encoding to examine their combined effect on behavioral recognition and associated ERPs. Participants performed eight experimental blocks consisting of an encoding phase and recognition test, with EEG recorded throughout. During encoding, pairs of objects and checkerboards were presented and participants were cued to attend to the left or right stream and detect targets as quickly as possible. In four blocks, stimulus presentation followed a rhythmic (constant, predictable) temporal structure, and in the other four blocks, stimulus onset was arrhythmic (random, unpredictable). An interaction between temporal expectation and spatial attention emerged, with greater recognition in the rhythmic than the arrhythmic condition for spatially attended items. Analysis of memory-specific ERP components uncovered effects of spatial attention. There were late positive component and FN400 old/new effects in the attended condition for both rhythmic and arrhythmic items, whereas in the unattended condition, there was an FN400 old/new effect and no late positive component effect. The study provides new evidence that memory improvement as a function of temporal expectation is dependent upon spatial attention.


Coupling-loss reduction of a vertically coupled microring resonator filter by spot-size-matched busline waveguides.
To improve the input-output coupling loss of a vertically coupled microring resonator filter, we fabricated microring resonators on an antiresonant reflecting optical waveguide (ARROW) with a large spot size and on the rectangular busline waveguide with a spot-size transformer. The spot size and the tapered structure were optimally designed from the viewpoint of spot-size matching to single-mode fibers and the reduction of radiation loss. Clear dropping responses were demonstrated for the ARROW-based microring resonator filters, and the coupling loss was successfully reduced by 22 dB.


The accuracy and cost-effectiveness of strategies used to identify peripheral artery disease among patients with diabetic foot ulcers.
Patients with diabetic foot ulcers (DFUs) should be evaluated for peripheral artery disease (PAD). We sought to estimate the overall diagnostic accuracy for various strategies that are used to identify PAD in this population. A Markov model with probabilistic and deterministic sensitivity analyses was used to simulate the clinical events in a population of 10,000 patients with diabetes. One of 14 different diagnostic strategies was applied to those who developed DFUs. Baseline data on diagnostic accuracy of individual noninvasive tests were based on a meta-analysis of previously reported studies. The overall sensitivity and cost-effectiveness of the 14 strategies were then compared. The overall sensitivity of various combinations of diagnostic testing strategies ranged from 32.6% to 92.6%. Cost-effective strategies included ankle-brachial indices for all patients; skin perfusion pressures (SPPs) or toe-brachial indices (TBIs) for all patients; and SPPs or TBIs to corroborate normal pulse examination findings, a strategy that lowered leg amputation rates by 36%. Strategies that used noninvasive vascular testing to investigate only abnormal pulse examination results had low overall diagnostic sensitivity and were weakly dominated in cost-effectiveness evaluations. Population prevalence of PAD did not alter strategy ordering by diagnostic accuracy or cost-effectiveness. TBIs or SPPs used uniformly or to corroborate a normal pulse examination finding are among the most sensitive and cost-effective strategies to improve the identification of PAD among patients presenting with DFUs. These strategies may significantly reduce leg amputation rates with only modest increases in cost.


Impact of Stress Reduction Interventions on Hostility and Ambulatory Systolic Blood Pressure in African American Adolescents.
This study examined the impact of breathing awareness meditation (BAM), life skills (LS) training, and health education (HE) interventions on self-reported hostility and 24-hour ambulatory blood pressure (ABP) in 121 African American (AA) ninth graders at increased risk for development of essential hypertension. They were randomly assigned to BAM, LS, or HE and engaged in intervention sessions during health class for 3 months. Before, after, and 3 months following intervention cessation, self-reported hostility and 24-hour ABP were measured. Results indicated that between pre- and postintervention, BAM participants displayed significant reductions in self-reported hostility and 24-hour systolic ABP. Reductions in hostility were significantly related to reductions in 24-hour systolic ABP. Between postintervention and follow-up, participants receiving LS showed a significant reduction in hostility but not in 24-hour ABP. Significant changes were not found for the HE group in 24-hour ABP or self-reported hostility, but these change scores were significantly correlated. The implications of the findings are discussed with regard to behavioral stress reduction programs for the physical and emotional health of AAs.


NKG2D-RAE-1 receptor-ligand variation does not account for the NK cell defect in nonobese diabetic mice.
NK cells from NOD mice induced with poly(I:C) in vivo exhibit low cytotoxicity against a range of target cells, but the genetic mechanisms controlling this defect are yet to be elucidated. Defects in the expression of NKG2D and its ligands, the RAE-1 molecules, have been hypothesized to contribute to the reduced NK function present in NOD mice. In this study, we show that segregation of the NK-mediated killing phenotype did not correlate with the NOD Raet1 haplotype and that the large alterations in NKG2D expression previously reported on NK cells expanded in vitro were not observed in primary, poly(I:C)-elicited NK cells in vivo. Additional studies indicate a complex genetic control of defective NOD NK cells including genes linked to the MHC and possibly those that are associated with an altered cytokine response to the TLR3-agonist poly(I:C).


Emissions of inorganic and organic arsenic compounds via the leachate pathway from pretreated municipal waste materials: a landfill reactor study.
The emission of arsenic (As) with leachate from mechanically biologically pretreated municipal solid waste (MBP-MSW) was quantified over one year using landfill simulation reactors. Arsenic mobilization and transformation processes were studied by simulating different environmental conditions (anoxic conditions with underlying soil or oxic/anoxic conditions). Amounts of mono-, di-, and trimethylated As in MBP-MSW prior to simulation were < 48 microg As kg(-1) and were magnified to 300-390 microg As kg(-1) under anoxic conditions, whereas methylated As was undetectable in the oxic setup. The highest leachate concentrations (up to 84 microg L(-1)) occurred during the first four weeks of manipulation. The annual Astotal release with leachates averaged 19.6, 7.6, and 4.5 microg kg(-1) under an anoxic environment with underlying soil, oxic conditions, and anoxic conditions, respectively, with 15-50% occurring as organic As. The annually released As represented 0.2-0.8% of the Astotal pool, suggesting that As mobilization from waste is a slow process. The anoxia diminished As release rates, whereas anoxic conditions with underlying soil material elevated the As mobilization, probably due to reductive dissolution of soil-derived Fe and Mn (hydr)oxides. The mass balance of methylated As in MBP-MSW and leachates before and after the treatments highlights As methylation under anoxic conditions and demethylation under oxic landfill conditions.


Psychophysical responses in patients receiving a mock laser within context of an acupuncture clinical trial: an interoceptive perspective.
The psychophysical responses induced by verum acupuncture are characterized by a constellation of unique subjective sensory responses commonly termed De Qi. Furthermore, a variety of sham interventions have been used as a control for acupuncture clinical trials. Indeed, one such control has been mock laser which has been used as control intervention in several acupuncture clinical controlled trials. The current study aim was to examine the De Qi sensory responses and its related characteristics elicited from acupuncture and compare them to those reported following sham laser in participants enrolled in a clinical trial. The study was embedded in a multi-center, two-arm randomised clinical trial, which evaluated the effect of acupuncture on lateral elbow pain. De Qi was assessed using the Massachusetts General Hospital Acupuncture Sensation Scale (MASS). Ninety-six participants were randomly allocated to receive either acupuncture (n = 47) or mock laser (n = 49) at the acupoints LI 10 and LI 11. Participants in both intervention groups reported similar De Qi psychophysical characteristics; however, both intensity and frequency of the individually perceived De Qi characteristics were significantly higher in the acupuncture group. 'Soreness', 'deep pressure', and 'fullness-distension' in the acupuncture group and 'tingling', and 'sharp pain' in mock laser group, were identified as the leading characteristics. Similar level of MASS De Qi Index (MDI) scores were reported for 'Hong Kong-China' and 'Australia-Italy' with a significantly higher level of De Qi reported by 'Hong Kong-China'. Furthermore, two distinct De Qi categories were identified, namely De Qi (in line with classical sensory responses of Suan, Ma, Zhang, and Zhong) and pain. Subjective 'somatic or interoceptive awareness' should be taken into account when De Qi psychophysical responses are examined. The study accentuates the necessity and the significance of further research into interoception phenomenon which may contribute to a better understanding of the placebo effect and De Qi psychophysical responses. Australian and New Zealand Clinical Trial Registry reference: ACTRN12613001138774 on 11th of October 2013.


A self-organizing neural network architecture for navigation using optic flow.
This article describes a self-organizing neural network architecture that transforms optic flow and eye position information into representations of heading, scene depth, and moving object locations. These representations are used to navigate reactively in simulations involving obstacle avoidance and pursuit of a moving target. The network's weights are trained during an action-perception cycle in which self-generated eye and body movements produce optic flow information, thus allowing the network to tune itself without requiring explicit knowledge of sensor geometry. The confounding effect of eye movement during translation is suppressed by learning the relationship between eye movement outflow commands and the optic flow signals that they induce. The remaining optic flow field is due to only observer translation and independent motion of objects in the scene. A self-organizing feature map categorizes normalized translational flow patterns, thereby creating a map of cells that code heading directions. Heading information is then recombined with translational flow patterns in two different ways to form maps of scene depth and moving object locations. Most of the learning processes take place concurrently and evolve through unsupervised learning. Mapping the learned heading representations onto heading labels or motor commands requires additional structure. Simulations of the network verify its performance using both noise-free and noisy optic flow information.


Familial Hypercholesterolemia in the Arabian Gulf Region: Clinical results of the Gulf FH Registry.
Familial hypercholesterolemia (FH) is a common autosomal dominant disorder that can result in premature atherosclerotic cardiovascular disease (ASCVD). Limited data are available worldwide about the prevalence and management of FH. Here, we aimed to estimate the prevalence and management of patients with FH in five Arabian Gulf countries (Saudi Arabia, Oman, United Arab Emirates, Kuwait, and Bahrain). The multicentre, multinational Gulf FH registry included adults (≥18 years old) recruited from outpatient clinics in 14 tertiary-care centres across five Arabian Gulf countries over the last five years. The Gulf FH registry had four phases: 1- screening, 2- classification based on the Dutch Lipid Clinic Network, 3- genetic testing, and 4- follow-up. Among 34,366 screened patient records, 3713 patients had suspected FH (mean age: 49±15 years; 52% women) and 306 patients had definite or probable FH. Thus, the estimated FH prevalence was 0.9% (1:112). Treatments included high-intensity statin therapy (34%), ezetimibe (10%), and proprotein convertase subtilisin/kexin type 9 inhibitors (0.4%). Targets for low-density lipoprotein cholesterol (LDL-C) and non-high-density lipoprotein cholesterol were achieved by 12% and 30%, respectively, of patients at high ASCVD risk, and by 3% and 6%, respectively, of patients at very high ASCVD risk (p <0.001; for both comparisons). This snap-shot study was the first to show the high estimated prevalence of FH in the Arabian Gulf region (about 3-fold the estimated prevalence worldwide), and is a "call-to-action" for further confirmation in future population studies. The small proportions of patients that achieved target LDL-C values implied that health care policies need to implement nation-wide screening, raise FH awareness, and improve management strategies for FH.


Advanced Glycation Endproducts and Bone Material Properties in Type 1 Diabetic Mice.
Fractures, particularly at the lower extremities and hip, are a complication of diabetes. In both type 1 (T1D) and type 2 diabetes (T2D), fracture risk is disproportionately worse than that predicted from the measurement of bone mineral density. Although an explanation for this discrepancy is the presence of organic matrix abnormalities, it has not been fully elucidated how advanced glycation endproducts (AGEs) relate to bone deterioration at both the macroscopic and microscopic levels. We hypothesized that there would be a relationship between skeletal AGE levels (determined by Raman microspectroscopy at specific anatomical locations) and bone macroscopic and microscopic properties, as demonstrated by the biomechanical measures of crack growth and microindentation respectively. We found that in OVE26 mice, a transgenic model of severe early onset T1D, AGEs were increased by Raman (carboxymethyl-lysine [CML] wildtype (WT): 0.0143 ±0.0005 vs T1D: 0.0175 ±0.0002, p = 0.003) at the periosteal surface. These differences were associated with less tough bone in T1D by fracture mechanics (propagation toughness WT: 4.73 ± 0.32 vs T1D: 3.39 ± 0.24 NM/m1/2, p = 0.010) and by reference point indentation (indentation distance increase WT: 6.85 ± 0.44 vs T1D: 9.04 ± 0.77 μm; p = 0.043). Within T1D, higher AGEs by Raman correlated inversely with macroscopic bone toughness. These data add to the existing body of knowledge regarding AGEs and the relationship between skeletal AGEs with biomechanical indices.


ONIOM approach for non-adiabatic on-the-fly molecular dynamics demonstrated for the backbone controlled Dewar valence isomerization.
Non-adiabatic on-the-fly molecular dynamics (NA-O-MD) simulations require the electronic wavefunction, energy gradients, and derivative coupling vectors in every timestep. Thus, they are commonly restricted to the excited state dynamics of molecules with up to ≈20 atoms. We discuss an approximation that combines the ONIOM(QM:QM) method with NA-O-MD simulations to allow calculations for larger molecules. As a proof of principle we present the excited state dynamics of a (6-4)-lesion containing dinucleotide (63 atoms), and especially the importance to include the confinement effects of the DNA backbone. The method is able to include electron correlation on a high level of theory and offers an attractive alternative to QM:MM approaches for moderate sized systems with unknown force fields.


CYP17A1: a biochemistry, chemistry, and clinical review.
Cytochrome P450 17A1 (CYP17A1; also P450c17and P450sccII) is a critically important enzyme in humans that catalyzes the formation of all endogenous androgens. It is an atypical cytochrome P450 enzyme in that it catalyzes two distinct types of substrate oxidation. Through its hydroxylase activity, it catalyzes the 17α-hydroxylation of pregnenolone to 17α-OH pregnenolone. Subsequently, through its C17,20lyase activity, it can further convert 17α-OH pregnenolone to the androgen dehydroepiandrosterone, which is a precursor to androstenedione, testosterone, and dihydrotestosterone. The importance of androgens in diseases such as prostate cancer has been appreciated for decades and the discovery of extra-testicular formation of androgens has helped clarify the pathology of the disease, especially the castrate- resistant disease. Therefore, specific inhibition of CYP17A1 by therapeutic intervention has been an area of considerable effort in several research laboratories. This basic research has led to the discovery of several promising drug candidates followed by the conduct of several clinical trials. Recently, all these efforts have culminated in the first approval by FDA of an inhibitor of CYP17A1 for the treatment of castrate-resistant prostate cancer. Ongoing clinical trials are now evaluating the agent in earlier stages of prostate cancer and even rare forms of androgen-dependent breast cancer. Accordingly, this review focuses on the biochemistry, chemistry, and clinical inhibitors of CYP17A1.


Suicidality and Associated Factors Among Men Who Have Sex With Men in Estonia.
Many studies over the last decade provide strong evidence of elevated rates of suicidality among lesbian, gay, bisexual, and transgender populations compared to heterosexual populations. The aim of the current study was to investigate suicidal ideation and attempts and related factors among men who have sex with men (MSM) in Estonia. We conducted a nationwide Internet-based study among MSM with 265 eligible participants. Our results reveal high rates of self-reported suicidal ideation (lifetime prevalence 45%) and attempts (lifetime prevalence 11%) as well as a high rate of problem drinking (33%) and depressive symptoms (32%) among MSM. Recent suicidal ideation (14% had thought about suicide in the last 12 months) was related to depressive symptoms and lower socioeconomic situation. There is a need to develop specific interventions to address these issues among MSM and to provide gay-friendly mental health services.


Multi-drug resistance of Escherichia coli from the urinary tract.
Resistance to antimicrobials of different structural classes has arisen in a multitude of bacterial species. This may complicate the therapeutic management of infections, including those of the urinary tract. The aim of this study is to show that antibiotic resistance surveillance is essential. A total of 11,126 urine samples were received at Sultan Qaboos University Hospital (SQUH), Muscat, Sultanate of Oman, for one year period (2001), they were cultured for bacterial pathogen and sensitivity was performed for all significant isolates. Eight hundred and eighty-four samples were culture positive with Escherichia coli (E.coli) (N=445) as the most common enterobacteriaceae, followed by Klebsiella pneumoniae (N=161), unidentified Coliforms (N=56) and Klebsiella species (N=33). High rate of resistance to b-Lactams was seen for all Enterobacteriaceae, maximum resistance was against ampicillin (78.6%) followed by co-amoxiclav (44.6%), but cefuroxime demonstrated the least resistance. Resistance to quinolone ranged from 4.7%-21.2% for all Enterobacteriaceae, cotrimoxazole resistance for all Enterobacteriaceae was 32.1%. Multi-drug resistance of E.coli was analyzed, 11.2% (50 of 445) were resistant to 3 or more antimicrobials and considered multidrug resistant. Among the multidrug-resistant isolates, 88% were resistant co-amoxiclav or cotrimoxazole, gentamicin 62%, ciprofloxacin 66%, and nitrofurantoin 32%. The predominant phenotype among multidrug-resistant isolates (26%; 13 of 50) included resistance to co-amoxiclav, gentamicin, and cotrimoxazole. This was the most common phenotype followed by 20%, of co-amoxiclav, cotrimoxazole and ciprofloxacin. In view of the current prevalence of multi-drug resistance among urinary tract isolates of E.coli at the SQUH (11.2%), continued surveillance will be beneficial.


Tumorigenicity of cancer stem-like cells derived from hepatocarcinoma is regulated by microRNA-145.
microRNAs are implicated in cancer initiation and progression by their ability to affect the expression of genes and proteins that regulate cell proliferation and death. Recent studies found that the stem cell-related genes Sox2, Oct4 and Klf4 are among the target genes regulated by microRNA-145 (miR-145), suggesting that miR-145 possibly plays a role in the maintenance of cancer stem cells. Therefore, it is important to address the involvement of miR-145 in the key roles of cancer stem cells in cancer initiation, progression and reoccurrence. We compared miR-145 expression in the cancer stem-like cells (T3A-A3) derived from hepatocarcinoma, in the hepatocarcinoma cell line BEL-7402 and in the normal liver sinusoidal endothelial cell line (LSEC). As demonstrated by a TaqMan microRNA real-time assay, T3A-A3 cells express lower miR-145 levels compared to the other cell lines. To address the role of miR-145 in cancer stem cells, miR-145 was restored in T3A-A3 cells. This resulted in senescence-like G1 arrest in cell cycling, and significantly inhibited clonogenic cell expansion in vitro and xenograft tumor growth in vivo. Moreover, miR-145 restoration diminished tumorsphere growth of T3A-A3 cells in vitro and T3A-A3 cells tumor formation in nude mice in vivo. Additionally, the increase in miR-145 levels paralleled the decrease in Oct4 levels. The effect of miR-145 on tumor suppression in T3A-A3 cells was partly reversed by overexpression of Oct4 both in vitro and in vivo. Collectively, our data indicate that miR-145 plays an important role in cancer stem cell tumorigenicity, potentially via modulation of the downstream target, Oct4.


Normally unused positional cues guide ectopic afferents in the leech CNS.
Central projections from peripheral sensory neurons segregate into distinct, ventrally positioned longitudinal tracts within the segmental ganglia of the leech Hirudo medicinalis. As documented here, there is an additional tract in the neuropils of the fifth and sixth body ganglia, located at the lateral margin and formed by afferent axons (the "sex afferents") originating from sensory neurons located in the male and female genitalia. Ablation of the genitalia results in the complete absence of this additional tract. We asked (1) whether segmental differences exist in the distribution of pathway cues available to the sex afferents, and (2) whether central pathway selection by these axons is specific. We transplanted the primordia of the male genitalia to several ectopic positions posterior to the sixth body segment and labeled the ectopic sex afferents in order to examine their paths in the CNS. In about 50% of the experimental animals, afferent axons originating in the transplanted tissue segregated into a distinct lateral fiber bundle within the neuropil of a nearby ganglion, in a position corresponding to their normal one in the sex ganglia. The sex afferents therefore find their normal pathways even in the ganglia of inappropriate segments, although these pathways are not used by any other afferents in these ganglia. We propose, therefore, that the positional cues employed by afferent axons to select appropriate pathways in the ganglionic neuropil are expressed in all segments of the leech CNS, regardless of whether such cues are normally used by afferent axons in each segment.


Prevalence of hypothyroidism in patients with polymyalgia rheumatica and giant cell arteritis.
The prevalence of thyroid disease and thyroid autoantibodies was evaluated in 367 patients with polymyalgia rheumatica (PMR) and/or giant cell arteritis (GCA). Thirty-seven patients had antibodies to thyroid microsomes or thyroglobulin; 18 had hypothyroidism requiring thyroxine replacement therapy. The prevalence of hypothyroidism (4.9%) was significantly greater than that found in 84 control subjects.


Lentinan has a beneficial effect on cognitive deficits induced by chronic Toxoplasma gondii infection in mice.
Toxoplasma gondii (T. gondii) is increasingly considered a risk factor for neurodegenerative diseases. However, there is only limited information on the development of drugs for T. gondii infection. Lentinan from Lentinula edodes is a bioactive ingredient with the potential to enhance anti-infective immunity. The present study aimed to investigate the neuroprotective effect of lentinan on T. gondii-associated cognitive deficits in mice. A chronic T. gondii infection mouse model was established by administering 10 cysts of T. gondii by gavage. Lentinan was intraperitoneally administered 2 weeks before infection. Behavioral tests, RNA sequencing, immunofluorescence, transmission electron microscopy and Golgi-Cox staining were performed to assess the effect of lentinan on cognitive deficits and neuropathology in vivo. In vitro, the direct and indirect effects of lentinan on the proliferation of T. gondii tachyzoites were evaluated in the absence and presence of BV-2 cells, respectively. Lentinan prevented T. gondii-induced cognitive deficits and altered the transcriptome profile of genes related to neuroinflammation, microglial activation, synaptic function, neural development and cognitive behavior in the hippocampus of infected mice. Moreover, lentinan reduced the infection-induced accumulation of microglia and downregulated the mRNA expression of proinflammatory cytokines. In addition, the neurite and synaptic ultrastructural damage in the hippocampal CA1 region due to infection was ameliorated by lentinan administration. Lentinan decreased the cyst burden in the brains of infected mice, which was correlated with behavioral performance. In line with this finding, lentinan could significantly inhibit the proliferation of T. gondii tachyzoites in the microglial cell line BV2, although lentinan had no direct inhibitory effect on parasite growth. Lentinan prevents cognitive deficits via the improvement of neurite impairment and synaptic loss induced by T. gondii infection, which may be associated with decreased cyst burden in the brain. Overall, our findings indicate that lentinan can ameliorate T. gondii-related neurodegenerative diseases.


An in vivo assay for the reverse transcriptase of human retrotransposon L1 in Saccharomyces cerevisiae.
L1 elements constitute a highly repetitive human DNA family (50,000 to 100,000 copies) lacking long terminal repeats and ending in a poly(A) tail. Some L1 elements are capable of retrotransposition in the human genome (Kazazian, H. H., Jr., C. Wong, H. Youssoufian, A. F. Scott, D. G. Phillips, and S.E. Antonarakis, Nature (London) 332:164-166, 1988). Although most are 5' truncated, a consensus sequence of complete L1 elements is 6 kb long and contains two open reading frames (ORFs) (Scott, A. F., B. J. Schmeckpeper, M. Abdelrazik, C. T. Comey, B. O'Hara, J. P. Rossiter, T. Cooley, P. Health, K. D. Smith, and L. Margolet, Genomics 1:113-125, 1987). The protein encoded by ORF2 has reverse transcriptase (RT) activity in vitro (Mathias, S. L., A. F. Scott, H. H. Kazazian, Jr., J. D. Boeke, and A. Gabriel, Science 254:1808-1810, 1991). Because L1 elements are so numerous, efficient methods for identifying active copies are required. We have developed a simple in vivo assay for the activity of L1 RT based on the system developed by Derr et al. (Derr, L. K., J. N. Strathern, and D. J. Garfinkel, Cell 67:355-364, 1991) for yeast HIS3 pseudogene formation. L1 ORF2 displays an in vivo RT activity similar to that of yeast Ty1 RT in this system and generates pseudogenes with unusual structures. Like the HIS3 pseudogenes whose formation depends on Ty1 RT, the HIS3 pseudogenes generated by L1 RT are joined to Ty1 sequences and often are part of complex arrays of Ty1 elements, multiple HIS3 pseudogenes, and hybrid Ty1/L1 elements. These pseudogenes differ from those previously described in that there are base pairs of unknown origin inserted at several of the junctions. In two of three HIS3 pseudogenes studied, the L1 RT appears to have jumped from the 5' end of a Ty1/L1 transcript to the poly(A) tract of the HIS3 RNA.


Psychological therapies for the management of chronic pain (excluding headache) in adults.
Chronic non-cancer pain, a disabling and distressing condition, is common in adults. It is a global public health problem and economic burden on health and social care systems and on people with chronic pain. Psychological treatments aim to reduce pain, disability and distress. This review updates and extends its previous version, published in 2012. To determine the clinical efficacy and safety of psychological interventions for chronic pain in adults (age > 18 years) compared with active controls, or waiting list/treatment as usual (TAU). We identified randomised controlled trials (RCTs) of psychological therapies by searching CENTRAL, MEDLINE, Embase and PsycINFO to 16 April 2020. We also examined reference lists and trial registries, and searched for studies citing retrieved trials. RCTs of psychological treatments compared with active control or TAU of face-to-face therapies for adults with chronic pain. We excluded studies of headache or malignant disease, and those with fewer than 20 participants in any arm at treatment end. Two or more authors rated risk of bias, extracted data, and judged quality of evidence (GRADE). We compared cognitive behavioural therapy (CBT), behavioural therapy (BT), and acceptance and commitment therapy (ACT) with active control or TAU at treatment end, and at six month to 12 month follow-up. We did not analyse the few trials of other psychological treatments. We assessed treatment effectiveness for pain intensity, disability, and distress. We extracted data on adverse events (AEs) associated with treatment. We added 41 studies (6255 participants) to 34 of the previous review's 42 studies, and now have 75 studies in total (9401 participants at treatment end). Most participants had fibromyalgia, chronic low back pain, rheumatoid arthritis, or mixed chronic pain. Most risk of bias domains were at high or unclear risk of bias, with selective reporting and treatment expectations mostly at unclear risk of bias. AEs were inadequately recorded and/or reported across studies. CBT The largest evidence base was for CBT (59 studies). CBT versus active control showed very small benefit at treatment end for pain (standardised mean difference (SMD) -0.09, 95% confidence interval (CI) -0.17 to -0.01; 3235 participants; 23 studies; moderate-quality evidence), disability (SMD -0.12, 95% CI -0.20 to -0.04; 2543 participants; 19 studies; moderate-quality evidence), and distress (SMD -0.09, 95% CI -0.18 to -0.00; 3297 participants; 24 studies; moderate-quality evidence). We found small benefits for CBT over TAU at treatment end for pain (SMD -0.22, 95% CI -0.33 to -0.10; 2572 participants; 29 studies; moderate-quality evidence), disability (SMD -0.32, 95% CI -0.45 to -0.19; 2524 participants; 28 studies; low-quality evidence), and distress (SMD -0.34, 95% CI -0.44 to -0.24; 2559 participants; 27 studies; moderate-quality evidence). Effects were largely maintained at follow-up for CBT versus TAU, but not for CBT versus active control. Evidence quality for CBT outcomes ranged from moderate to low. We rated evidence for AEs as very low quality for both comparisons. BT We analysed eight studies (647 participants). We found no evidence of difference between BT and active control at treatment end (pain SMD -0.67, 95% CI -2.54 to 1.20, very low-quality evidence; disability SMD -0.65, 95% CI -1.85 to 0.54, very low-quality evidence; or distress SMD -0.73, 95% CI -1.47 to 0.01, very low-quality evidence). At follow-up, effects were similar. We found no evidence of difference between BT and TAU (pain SMD -0.08, 95% CI -0.33 to 0.17, low-quality evidence; disability SMD -0.02, 95% CI -0.24 to 0.19, moderate-quality evidence; distress SMD 0.22, 95% CI -0.10 to 0.54, low-quality evidence) at treatment end. At follow-up, we found one to three studies with no evidence of difference between BT and TAU. We rated evidence for all BT versus active control outcomes as very low quality; for BT versus TAU. Evidence quality ranged from moderate to very low. We rated evidence for AEs as very low quality for BT versus active control. No studies of BT versus TAU reported AEs. ACT We analysed five studies (443 participants). There was no evidence of difference between ACT and active control for pain (SMD -0.54, 95% CI -1.20 to 0.11, very low-quality evidence), disability (SMD -1.51, 95% CI -3.05 to 0.03, very low-quality evidence) or distress (SMD -0.61, 95% CI -1.30 to 0.07, very low-quality evidence) at treatment end. At follow-up, there was no evidence of effect for pain or distress (both very low-quality evidence), but two studies showed a large benefit for reducing disability (SMD -2.56, 95% CI -4.22 to -0.89, very low-quality evidence). Two studies compared ACT to TAU at treatment end. Results should be interpreted with caution. We found large benefits of ACT for pain (SMD -0.83, 95% CI -1.57 to -0.09, very low-quality evidence), but none for disability (SMD -1.39, 95% CI -3.20 to 0.41, very low-quality evidence), or distress (SMD -1.16, 95% CI -2.51 to 0.20, very low-quality evidence). Lack of data precluded analysis at follow-up. We rated evidence quality for AEs to be very low. We encourage caution when interpreting very low-quality evidence because the estimates are uncertain and could be easily overturned. We found sufficient evidence across a large evidence base (59 studies, over 5000 participants) that CBT has small or very small beneficial effects for reducing pain, disability, and distress in chronic pain, but we found insufficient evidence to assess AEs. Quality of evidence for CBT was mostly moderate, except for disability, which we rated as low quality. Further trials may provide more precise estimates of treatment effects, but to inform improvements, research should explore sources of variation in treatment effects. Evidence from trials of BT and ACT was of moderate to very low quality, so we are very uncertain about benefits or lack of benefits of these treatments for adults with chronic pain; other treatments were not analysed. These conclusions are similar to our 2012 review, apart from the separate analysis of ACT.


Mycobacteria-Specific Cytokine Responses Detect Tuberculosis Infection and Distinguish Latent from Active Tuberculosis.
Current immunodiagnostic tests for tuberculosis (TB), including the tuberculin skin test and IFN-γ release assay (IGRA), have significant limitations, which include their inability to distinguish between latent TB infection (LTBI) and active TB, a distinction critical for clinical management. To identify mycobacteria-specific cytokine biomarkers that characterize TB infection, determine their diagnostic performance characteristics, and establish whether these biomarkers can distinguish between LTBI and active TB. A total of 149 children investigated for TB infection were recruited; all participants underwent a tuberculin skin test and QuantiFERON-TB Gold assay. In parallel, whole-blood assays using early secretory antigenic target-6, culture filtrate protein-10, and PPD as stimulatory antigens were undertaken, and cytokine responses were determined by xMAP multiplex assays. IFN-γ, interferon-inducible protein-10 (IP-10), tumor necrosis factor (TNF)-α, IL-1ra, IL-2, IL-13, and MIP-1β (macrophage inflammatory protein-1β) responses were significantly higher in LTBI and active TB cases than in TB-uninfected individuals, irrespective of the stimulant. Receiver operating characteristic analyses showed that IP-10, TNF-α, and IL-2 responses achieved high sensitivity and specificity for the distinction between TB-uninfected and TB-infected individuals. TNF-α, IL-1ra, and IL-10 responses had the greatest ability to distinguish between LTBI and active TB cases; the combinations of TNF-α/IL-1ra and TNF-α/IL-10 achieved correct classification of 95.5% and 100% of cases, respectively. We identified several mycobacteria-specific cytokine biomarkers with the potential to be exploited for immunodiagnosis. Incorporation of these biomarkers into future immunodiagnostic assays for TB could result in substantial gains in sensitivity and allow the distinction between LTBI and active TB based on a blood test alone.


'It was like being hit with a brick': a qualitative study on the effect of clinicians' delivery of a diagnosis of eye disease for patients in primary and secondary care.
To explore patients' experiences of getting a diagnosis of eye disease, the psychological impact of this and how this could be improved. An exploratory qualitative interview study using a narrative approach and inductive methods. This study was conducted with patients who had attended ophthalmic appointments in primary and secondary care and in opticians located in the South of England. 18 people diagnosed with eye disease in England. Four themes were identified: the convoluted process of being diagnosed, the impact of clinicians' words, the search for information and reflections on what could be improved. The prolonged wait for a definitive diagnosis was a source of frustration and anxiety for many patients. Professionals' words and tone when delivering a diagnosis sometimes affected a patient's view of their diagnosis and their later ability to come to terms with it. Patients were desperate for information, but many felt they were not provided with sufficient information at the time of diagnosis and did not know whether to trust information found online. Participants felt the provision of a hospital liaison service and/or counselling could mitigate the impact on patients and families. Interactions with clinicians can have a lasting impact on how a diagnosis is experienced and how well the patient is able to come to terms with their visual impairment. Receiving little or no information left patients feeling lost and unsupported. This led them to search for information from less reliable sources. Clinicians should consider how they communicate a diagnosis to patients, how and when they offer information about diagnosis and prognosis and where possible signpost patients to additional support systems and counselling services as early as possible.


Chromosome aberrations in lymphocytes of persons exposed to an earthquake in Armenia.
This study attempted to determine the level of chromosome aberrations in lymphocytes of victims of the 1988 earthquake in Armenia. Chromosome aberrations were measured in blood samples taken from 41 victims of the earthquake that hit Armenia in 1988 and in samples of 47 reference blood donors. The victims suffered from severe psychoemotional stress but were otherwise healthy. All the samples were taken 2 to 3 weeks after the earthquake. All the subjects were lifetime nonsmokers. The cells were scored blind as to the exposure status. The subjects exposed to the earthquake had a higher proportion of cells with chromosome aberrations [3.1 (SD 2.1)%] than the referents [1.7 (SD 1.3)%, P-value for the difference 0.0009]. The difference persisted when the values were adjusted for age and gender [relative risk (RR) 1.9, 95% confidence interval (95% CI) 1.4-2.5]. The difference was present for double breaks (RR 4.1, 95% CI 2.6-6.4), but not for single breaks (RR 1.1, 95% CI 0.8-1.7). The exposed subjects also had a lower percentage of cells with 46 chromosomes (P=0.03) than the referents. This study suggests an increase in chromosome aberrations in the lymphocytes of victims of a severe earthquake as compared with the levels of referents. If not due to bias or confounding, the difference may reflect the effect of either environmental exposures related to the earthquake or severe psychogenic stress. The levels of chromosome aberrations found among the earthquake victims in this study are comparable with those found in prospective studies of long-term cancer risk.


DMD and IL1RAPL1: two large adjacent genes localized within a common fragile site (FRAXC) have reduced expression in cultured brain tumors.
Common fragile sites (CFSs) are large regions of profound genomic instability found in all individuals. Spanning the center of the two most frequently expressed CFS regions, FRA3B (3p14.3) and FRA16D (16q23.2), are the 1.5 Mb FHIT gene and the 1.0 Mb WWOX gene. These genes are frequently deleted and/or altered in many different cancers. Both FHIT and WWOX have been demonstrated to function as tumor suppressors, both in vitro and in vivo. A number of other large CFS genes have been identified and are also frequently inactivated in multiple cancers. Based on these data, several additional very large genes were tested to determine if they were derived from within CFS regions, but DCC and RAD51L1 were not. However, the 2.0 Mb DMD gene and its immediately distal neighbor, the 1.8 Mb IL1RAPL1 gene are CFS genes contained within the FRAXC CFS region (Xp21.2-->p21.1). They are abundantly expressed in normal brain but were dramatically underexpressed in every brain tumor cell line and xenograft (derived from an intracranial model of glioblastoma multiforme) examined. We studied the expression of eleven other large CFS genes in the same panel of brain tumor cell lines and xenografts and found reduced expression of multiple large CFS genes in these samples. In this report we show that there is selective loss of specific large CFS genes in different cancers that does not appear to be mediated by the relative instability within different CFS regions. Further, the inactivation of multiple large CFS genes in xenografts and brain tumor cell lines may help to explain why this type of cancer is highly aggressive and associated with a poor clinical outcome.


Site-specific expression of transferrin receptor by human colon cancer cells directly correlates with eradication by antitransferrin recombinant immunotoxin.
We determined the efficacy of HB21(Fv)PE40, a single-chain immunotoxin made by fusing the variable regions of a monoclonal antibody directed at the human transferrin receptor (TfR) with a truncated mutant of Pseudomonas exotoxin (PE), against metastatic human colon carcinoma KM12L4 cells growing in the liver or subcutis of nude mice. Organ-specific modulation of TfR expression was examined by immunohistochemistry and flow cytometry using anti-human CD71 antibody. KM12L4 cells expressed human TfR and were lysed in vitro by HB21(Fv)PE40 but not LMB-7 (a control immunotoxin specific for a Lewis Y-related carbohydrate antigen). KM12L4 cells growing in the liver expressed higher levels of TfR than cells growing s.c. Systemic administration of HB21(Fv)PE40 eliminated KM12L4 liver metastasis, whereas administration of LMB-7 did not. Treatment of mice with HB21(Fv)PE40 only delayed the growth of s.c. tumors. KM12L4 cells recovered from liver metastases, expressed higher levels of TfR, and were more sensitive to lysis by HB21(Fv)PE40 than KM12L4 cells recovered from s.c. tumors. Indeed, collectively, the data show that the expression level of the TfR by human colon cancer cells is modulated by the organ microenvironment which can be advantageous for the use of therapeutic immunotoxins.


Tumour micro-environment elicits innate resistance to RAF inhibitors through HGF secretion.
Drug resistance presents a challenge to the treatment of cancer patients. Many studies have focused on cell-autonomous mechanisms of drug resistance. By contrast, we proposed that the tumour micro-environment confers innate resistance to therapy. Here we developed a co-culture system to systematically assay the ability of 23 stromal cell types to influence the innate resistance of 45 cancer cell lines to 35 anticancer drugs. We found that stroma-mediated resistance is common, particularly to targeted agents. We characterized further the stroma-mediated resistance of BRAF-mutant melanoma to RAF inhibitors because most patients with this type of cancer show some degree of innate resistance. Proteomic analysis showed that stromal cell secretion of hepatocyte growth factor (HGF) resulted in activation of the HGF receptor MET, reactivation of the mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3-OH kinase (PI(3)K)-AKT signalling pathways, and immediate resistance to RAF inhibition. Immunohistochemistry experiments confirmed stromal cell expression of HGF in patients with BRAF-mutant melanoma and showed a significant correlation between HGF expression by stromal cells and innate resistance to RAF inhibitor treatment. Dual inhibition of RAF and either HGF or MET resulted in reversal of drug resistance, suggesting RAF plus HGF or MET inhibitory combination therapy as a potential therapeutic strategy for BRAF-mutant melanoma. A similar resistance mechanism was uncovered in a subset of BRAF-mutant colorectal and glioblastoma cell lines. More generally, this study indicates that the systematic dissection of interactions between tumours and their micro-environment can uncover important mechanisms underlying drug resistance.


Bone marrow transplantation reveals an essential synergy between neuronal and hemopoietic cell neurokinin production in pulmonary inflammation.
Neurogenic inflammation is believed to originate with the antidromic release of substance P, and of other neurokinins encoded by the preprotachykinin A (PPT-A) gene, from unmyelinated nerve fibers (C-fibers) following noxious stimuli. Consistent with this concept, we show here that selective sensory-fiber denervation with capsaicin and targeted deletion of the PPT-A gene protect murine lungs against both immune complex-mediated and stretch-mediated injuries. Reconstitution of PPT-A gene-deleted mice with WT bone marrow does not abrogate this protection, demonstrating a critical role for PPT-A gene expression by sensory neurons in pulmonary inflammation. Surprisingly, reconstitution of WT mice with PPT-A gene-deficient bone marrow also confers protection against pulmonary injury, revealing that PPT-A gene expression in hemopoietic cells has a previously unanticipated essential role in tissue injury. Taken together, these findings demonstrate a critical synergy between capsaicin-sensitive sensory fibers and hemopoietic cells in neurokinin-mediated inflammation and suggest that such synergy may be the basis for a stereotypical mechanism of response to injury in the respiratory tract.


Requirement of Wnt/beta-catenin signaling in pronephric kidney development.
The pronephric kidney controls water and electrolyte balance during early fish and amphibian embryogenesis. Many Wnt signaling components have been implicated in kidney development. Specifically, in Xenopus pronephric development as well as the murine metanephroi, the secreted glycoprotein Wnt-4 has been shown to be essential for renal tubule formation. Despite the importance of Wnt signals in kidney organogenesis, little is known of the definitive downstream signaling pathway(s) that mediate their effects. Here we report that inhibition of Wnt/beta-catenin signaling within the pronephric field of Xenopus results in significant losses to kidney epithelial tubulogenesis with little or no effect on adjoining axis or somite development. We find that the requirement for Wnt/beta-catenin signaling extends throughout the pronephric primordium and is essential for the development of proximal and distal tubules of the pronephros as well as for the development of the duct and glomus. Although less pronounced than effects upon later pronephric tubule differentiation, inhibition of the Wnt/beta-catenin pathway decreased expression of early pronephric mesenchymal markers indicating it is also needed in early pronephric patterning. We find that upstream inhibition of Wnt/beta-catenin signals in zebrafish likewise reduces pronephric epithelial tubulogenesis. We also find that exogenous activation of Wnt/beta-catenin signaling within the Xenopus pronephric field results in significant tubulogenic losses. Together, we propose Wnt/beta-catenin signaling is required for pronephric tubule, duct and glomus formation in Xenopus laevis, and this requirement is conserved in zebrafish pronephric tubule formation.


Class Id ribonucleotide reductase utilizes a Mn2(IV,III) cofactor and undergoes large conformational changes on metal loading.
Outside of the photosynthetic machinery, high-valent manganese cofactors are rare in biology. It was proposed that a recently discovered subclass of ribonucleotide reductase (RNR), class Id, is dependent on a Mn2(IV,III) cofactor for catalysis. Class I RNRs consist of a substrate-binding component (NrdA) and a metal-containing radical-generating component (NrdB). Herein we utilize a combination of EPR spectroscopy and enzyme assays to underscore the enzymatic relevance of the Mn2(IV,III) cofactor in class Id NrdB from Facklamia ignava. Once formed, the Mn2(IV,III) cofactor confers enzyme activity that correlates well with cofactor quantity. Moreover, we present the X-ray structure of the apo- and aerobically Mn-loaded forms of the homologous class Id NrdB from Leeuwenhoekiella blandensis, revealing a dimanganese centre typical of the subclass, with a tyrosine residue maintained at distance from the metal centre and a lysine residue projected towards the metals. Structural comparison of the apo- and metal-loaded forms of the protein reveals a refolding of the loop containing the conserved lysine and an unusual shift in the orientation of helices within a monomer, leading to the opening of a channel towards the metal site. Such major conformational changes have not been observed in NrdB proteins before. Finally, in vitro reconstitution experiments reveal that the high-valent manganese cofactor is not formed spontaneously from oxygen, but can be generated from at least two different reduced oxygen species, i.e. H2O2 and superoxide (O 2·- ). Considering the observed differences in the efficiency of these two activating reagents, we propose that the physiologically relevant mechanism involves superoxide.


Short-term repeatability of measurements of antimicrobial susceptibility of Escherichia coli isolated from feces of feedlot cattle.
Short-term stability of measurements of antimicrobial susceptibility of Escherichia coli isolated from feces of feedlot cattle is important in developing monitoring and surveillance programs. Frequent evaluations (i.e., daily) are resource intensive and in some situations may be impractical for long-term sampling protocols. Consequently, a point-in-time measurement will need to be used to represent conditions in the perisampling period. In this study, 30 fecal samples were collected from each of 6 cattle pens on a commercial cattle feedlot on 2 occasions separated by 48 hours. Escherichia coli was isolated from single and pooled samples. The isolates were tested for antimicrobial susceptibility against a panel of 17 antimicrobials. Resistance to 5 antimicrobials (ampicillin, nalidixic acid, streptomycin, sulfamethoxazole, and tetracycline) was detected in single and pooled samples from both sampling periods (days 1 and 3). The prevalence of isolates resistant to these 5 antimicrobials was 2% or higher in all treatment combinations except for pools obtained from day 3 samples. Lower levels of resistance to 6 more antimicrobials were detected inconsistently across the single and pooled samples. Logistic models constructed for the antimicrobials to which the E. coli isolates were most commonly resistant demonstrated that there were no significant differences between periods (P > 0.10) and between single and pooled samples (P > 0.20). The distribution of the number of antimicrobials to which isolates were resistant was consistent for the single samples across periods, but there appeared to be a lower prevalence of any resistance in day 1 pooled samples. A larger number of resistant phenotypes were detected in the single samples than in the pooled samples, and resistant phenotypes with prevalence of less than 2% were detected inconsistently across periods and single and pooled samples. Resistance to individual antimicrobials was consistent by all measures when the prevalence was at least 2%. Inconsistent results were obtained for antimicrobials to which resistance rarely occurred. The apparent inconsistencies do not appear to be related to external factors but rather to sampling intensity. Short-term stability is a plausible assumption under sampling strategies that are designed to detect specific levels of prevalence. However, when resistance levels fall below these levels, there will likely be fluctuations in the presence or absence of rare resistant phenotypes and in their prevalence and central tendency measures.


Screening and mapping of pigments in paintings using scanning electrochemical microscopy (SECM).
The use of the scanning electrochemical microscopy (SECM) technique for identifying and mapping of both organic and inorganic pigments in sub-microsamples from pictorial specimens is described. This methodology, inspired by the voltammetry of immobilized particles technique, permits the study of textural properties of paint layers and mapping the distribution of pigment grains upon application of different potentials to the substrate. A combination of the redox competition SECM strategy with voltammetry yields a local identification methodology for different organic and inorganic pigments in paint samples.


Live imaging of collagen remodeling during angiogenesis.
To better understand interstitial matrix remodeling during angiogenesis, we probed endogenous optical signatures of collagen fibrils and cells with multiphoton microscopy to noninvasively visualize, in real-time, changes to fibril organization around angiogenic sprouts and growing neovessels. From analyses of the second-harmonic generation signal from fibrillar collagen and two-photon excited fluorescence, as well as coherent transmitted light from vascular cells, we found that microvessel fragments interacting with the collagen matrix exhibited two key features: a strong association of fibrillar collagen around the parent vessel fragment during vessel construct reconstitution and a substantial collagen fibril reorganization by sprout and neovessel tips. Results indicate that angiogenic sprouts and growing neovessels actively and differentially remodel existing collagen fibrils. This imaging approach to assess local changes in matrix organization may have a broader impact on tissue biology and mechanics during angiogenesis and allow for new insights in cardiovascular, diabetes, and cancer research.


Health economic evaluation of gene replacement therapies: methodological issues and recommendations.
Objective: To provide recommendations for addressing previously identified key challenges in health economic evaluations of Gene Replacement Therapies (GRTs), including: 1) the assessment of clinical effectiveness; 2) the valuation of health outcomes; 3) the time horizon and extrapolation of effects beyond trial duration; 4) the estimation of costs; 5) the selection of appropriate discount rates; 6) the incorporation of broader elements of value; and 7) affordability. Methods: A literature review on economic evaluations of GRT was performed. Interviews were conducted with 8 European and US health economic experts with experience in evaluations of GRT. Targeted literature reviews were conducted to investigate further potential solutions to specific challenges. Recommendations: Experts agreed on factors to be considered to ensure the acceptability of historical cohorts by HTA bodies. Existing prospective registries or, if not available, retrospective registries, may be used to analyse different disease trajectories and inform extrapolations. The importance of expert opinion due to limited data was acknowledged. Expert opinion should be obtained using structured elicitation techniques. Broader elements of value, beyond health gains directly related to treatment, can be considered through the application of a factor to inflate the quality-adjusted life years (QALYs) or a higher cost-effectiveness threshold. Additionally, the use of cost-benefit analysis and saved young life equivalents (SAVE) were proposed as alternatives to QALYs for the valuations of outcomes of GRT as they can incorporate broader elements of value and avoid problems of eliciting utilities for paediatric diseases. Conclusions: While some of the limitations of economic evaluations of GRT are inherent to limited clinical data and lack of experience with these treatments, others may be addressed by methodological research to be conducted by health economists.


Cerebrospinal Fluid Proteome Changes in Older Non-Cardiac Surgical Patients with Postoperative Cognitive Dysfunction.
Postoperative cognitive dysfunction (POCD), a syndrome of cognitive deficits occurring 1-12 months after surgery primarily in older patients, is associated with poor postoperative outcomes. POCD is hypothesized to result from neuroinflammation; however, the pathways involved remain unclear. Unbiased proteomic analyses have been used to identify neuroinflammatory pathways in multiple neurologic diseases and syndromes but have not yet been applied to POCD. To utilize unbiased mass spectrometry-based proteomics to identify potential neuroinflammatory pathways underlying POCD. Unbiased LC-MS/MS proteomics was performed on immunodepleted cerebrospinal fluid (CSF) samples obtained before, 24 hours after, and 6 weeks after major non-cardiac surgery in older adults who did (n = 8) or did not develop POCD (n = 6). Linear mixed models were used to select peptides and proteins with intensity differences for pathway analysis. Mass spectrometry quantified 8,258 peptides from 1,222 proteins in > 50%of patient samples at all three time points. Twelve peptides from 11 proteins showed differences in expression over time between patients with versus withoutPOCD (q < 0.05), including proteins previously implicated in neurodegenerative disease pathophysiology. Additionally, 283 peptides from 182 proteins were identified with trend-level differences (q < 0.25) in expression over time between these groups. Among these, pathway analysis revealed that 50 were from 17 proteins mapping to complement and coagulation pathways (q = 2.44*10-13). These data demonstrate the feasibility of performing unbiased mass spectrometry on perioperative CSF samples to identify pathways associated with POCD. Additionally, they provide hypothesis-generating evidence for CSF complement and coagulation pathway changes in patients with POCD.


Role of ultrafast-acting insulin analogues in the management of diabetes.
To control both fasting and prandial plasma glucose levels in people with diabetes, insulin therapy must mimic "normal" physiological insulin secretion as much as possible. This is achieved with a long-acting insulin injected once or twice daily and a bolus of insulin injected before every meal. Prandial (bolus) insulin can either be regular human insulin (RHI) or a rapid-acting insulin analogue (RAIA). Although the efficacy of RHI has been established over approximately 35 years of clinical use, RAIAs offer several clinical advantages over RHI, namely that they have been engineered with a reduced tendency to aggregate as hexamers, which allows for rapid dissociation and absorption after a subcutaneous injection. Conventional RAIAs include insulin lispro, insulin aspart, and insulin glulisine. The more recently developed fast-acting insulin aspart (faster aspart) is an ultrafast-acting mealtime insulin that contains the conventional insulin aspart in a new formulation with the excipients niacinamide and L-arginine to achieve faster insulin absorption than RHI and the conventional insulin aspart formulation. This article reviews the clinical evidence supporting the use of RAIAs as part of a basal-bolus regimen in patients with diabetes, with a focus on new formulations whose pharmacological profiles more closely mimic the endogenous prandial insulin secretion pattern that is seen in individuals without diabetes. This review also provides a clinical perspective to help guide health care professionals in the use of RAIAs.


Patched1 inhibits epidermal progenitor cell expansion and basal cell carcinoma formation by limiting Igfbp2 activity.
Basal cell carcinoma (BCC) of the skin is the most common form of cancer, with the majority being caused by mutations in the Patched1 (Ptch1) gene, leading to activation of the Hedgehog (Hh) signaling pathway. Hh signaling is implicated in many tumor types; thus, defining the mechanisms by which Ptch1 regulates tissue proliferation is of paramount importance. Here, we show that the key role of Ptch1 in the skin is to limit the size of the epidermal stem/progenitor compartment and allow hair follicle differentiation. Specifically, loss of Ptch1 leads to the promotion of progenitor cell fate by increasing basal cell proliferation and limiting the progression of basal cells into differentiated hair follicle cell types. Our data indicate that BCCs likely result from hair follicle progenitor cells that, due to Hh signal activation, cannot progress through normal hair follicle differentiation. These data confirm the role of Ptch1 as a negative regulator of epidermal progenitor turnover and also show for the first time that Ptch1 plays a role in the differentiation of the hair follicle lineage. In addition, we show that insulin-like growth factor binding protein 2 (Igfbp2) is upregulated in both murine and human BCCs and that blocking Igfbp2 activity reduces the Hh-mediated expansion of epidermal progenitor cells. We propose that Igfbp2 mediates epidermal progenitor cell expansion and therefore represents an epidermal progenitor cell-specific target of Hh signaling that promotes BCC development.


Termination of second-trimester pregnancy by mifepristone combined with misoprostol versus intra-amniotic injection of ethacridine lactate (Rivanol®): a systematic review of Chinese trials.
Mifepristone combined with misoprostol for second-trimester abortion (MM) and intra-amniotic injection of ethacridine lactate (EL; Rivanol®) are the common methods for termination of second-trimester pregnancy in China. The systematic review of relevant literature was conducted to evaluate the effectiveness and safety on termination of second-trimester pregnancy using MM, introduced in 1988 in China, versus the Chinese routinely used method since 1970 -- EL. The review was conducted to evaluate mifepristone combined with the misoprostol versus intra-amniotic injection of ethacridine lactate in China for termination of second-trimester pregnancy with respect to efficacy, side effects, complications and so on. The Cochrane Central Register of Controlled Trials, EMBASE, MEDLINE, POPLINE, TOXLINE, World Health Organization-Department of Reproductive Health and Research, Chinese Biomedical Literature Database, Chinese Journal Full-text Database and Chinese Science & Technology Journal Database were systematically searched. Reference lists for English and Chinese-language reports (published from 1966 to 2008) were searched. All randomized controlled trials (RCTs) on second-trimester abortion by combination of mifepristone and misoprostol versus ethacridine lactate, which conducted in China, were analyzed. Trial quality was assessed, and data extraction was made independently by two reviewers. Fifteen original RCTs using MM versus EL were included. Three trials used oral misoprostol, six trials used vaginal misoprostol, and six trials used oral plus vaginal misoprostol. Failure rate of abortion was 2.0% to 5.9% in the MM group and 7.4% to 20.7% in the EL group. The incidence of postabortion curettage was similar in the two groups. The time of labor and hospitalization for abortion in the MM group were shorter compared with EL group. The blood loss within 2 h of abortion in the MM group was significantly less than that in the EL group, but the blood loss within 24 h of abortion was the same in both groups. More gastrointestinal side effects occurred with the MM regimen, whereas cervical injury occurred more often in the EL group. Compared with the intra-amniotic injection of ethacridine lactate, mifepristone/misoprostol has a higher success rate and shorter time of labor with more gastrointestinal side effects for termination of second-trimester pregnancy.


Neural correlates of nouns and verbs in early bilinguals.
Previous neuroimaging research indicates that English verbs and nouns are represented in frontal and posterior brain regions, respectively. For Chinese monolinguals, however, nouns and verbs are found to be associated with a wide range of overlapping areas without significant differences in neural signatures. This different pattern of findings led us to ask the question of where nouns and verbs of two different languages are represented in various areas in the brain in Chinese-English bilinguals. In this study, we utilized functional magnetic resonance imaging (fMRI) and a lexical decision paradigm involving Chinese and English verbs and nouns to address this question. We found that while Chinese nouns and verbs involved activation of common brain areas, the processing of English verbs engaged many more regions than did the processing of English nouns. Specifically, compared to English nouns, English verb presentation was associated with stronger activation of the left putamen and cerebellum, which are responsible for motor function, suggesting the involvement of the motor system in the processing of English verbs. Our findings are consistent with the theory that neural circuits for linguistic dimensions are weighted and modulated by the characteristics of a language.