Bidirectional Mendelian randomisation analysis of the relationship between circulating vitamin D concentration and colorectal cancer risk.
Epidemiological evidence is consistent with a protective effect of vitamin D against colorectal cancer (CRC), but the observed strong associations are open to confounders and potential reverse causation. Previous Mendelian randomisation (MR) studies were limited by poor genetic instruments and inadequate statistical power. Moreover, whether genetically higher CRC risk can influence vitamin D level, namely the reverse causation, still remains unknown. Herein, we report the first bidirectional MR study. We employed 110 newly identified genetic variants as proxies for vitamin D to obtain unconfounded effect estimates on CRC risk in 26 397 CRC cases and 41 481 controls of European ancestry. To test for reserve causation, we estimated effects of 115 CRC-risk variants on vitamin D level among 417 580 participants from the UK Biobank. The causal association was estimated using the random-effect inverse-variance weighted (IVW) method. We found no significant causal effect of vitamin D on CRC risk [IVW estimate odds ratio: 0.97, 95% confidence interval (CI) = 0.88-1.07, P = .565]. Similarly, no significant reverse causal association was identified between genetically increased CRC risk and vitamin D levels (IVW estimate β: -0.002, 95% CI = -0.008 to 0.004, P = .543). Stratified analysis by tumour sites did not identify significant causal associations in either direction between vitamin D and colon or rectal cancer. Despite the improved statistical power of this study, we found no evidence of causal association of either direction between circulating vitamin D and CRC risk. Significant associations reported by observational studies may be primarily driven by unidentified confounders.


Transcatheter Aortic Valve Implantation: First Applications and Short Term Outcomes in Our Clinic.
The objective of this study is to evaluate the first applications and short term outcomes of transcatheter aortic valve implantation (TAVI) in our clinic, which is a new technology for the patients with high risk for surgical aortic valve replacement (SAVR). Between January 2010 and December 2012, twenty five patients (16 males, 9 females; mean age 74.04±8.86 years) diagnosed with severe aortic stenosis, who were at high risk for surgery (EuroSCORE II: 5.58±4.20) and underwent TAVI in our clinic, were evaluated. The demographic and clinical characteristics of patients, anaesthetic management, complications during pre- and post-operative periods and the mortality rate in the first 30 days and six months were recorded. Edwards SAPIEN Valve prostheses were implanted by transfemoral approach (percutaneously in 10 patients and surgically in 15 patients) in all patients. The TAVI procedure was performed under general anaesthesia. The success rate of the TAVI procedure was 100%. Three patients had limited dissection of the femoral artery; however, intervention was not needed due to good distal perfusion rate. Permanent pacemaker was implanted to four patients because of long-term atrioventricular blockage. After the procedure, all patients were transferred to the Intensive Care Unit (ICU) and all patients were extubated in the ICU. The mean mechanical ventilation duration (minutes) was 166.20±39.32, the mean critical care unit stay (day) was 5.64±2.99 and the mean hospital stay (day) was 11.92±5.54. Acute renal failure was observed in one patient and stroke was observed in two patients on the first postoperative day. The mortality rate in the first 30 days and 6 months was found to be 4% and 16%, respectively. Transcatheter aortic valve implantation is a great option for patients with severe aortic stenosis who are at high risk for SAVR. In our institute, procedural success and short term outcomes for patients underwent TAVI were found to be similar to the other studies in the national and international literature.


Implementation of strategies to prevent and control the emergence and spread of antimicrobial-resistant microorganisms in U.S. hospitals.
To examine the extent to which the strategies recommended by the National Foundation for Infectious Diseases (NFID)-Centers for Disease Control and Prevention (CDC) co-sponsored workshop, Antimicrobial Resistance in Hospitals: Strategies to Improve Antimicrobial Use and Prevent Nosocomial Transmission of Antimicrobial-Resistant Microorganisms, have been implemented and the relationship between the degree of implementation and hospital culture, leadership, and organizational factors. Survey. A representative sample of U.S. hospitals stratified by teaching status, bed size, and geographic region. Infection control professionals. Surveyed hospitals had implemented strategies to optimize the use of antimicrobials and to detect, report, and prevent transmission of antimicrobial-resistant microorganisms. Multivariate analyses found that hospitals with a greater degree of implementation of the NFID-CDC strategic goals were more likely to have management support, education of staff, and interdisciplinary groups specifically to address these issues; they were also more likely to engage in benchmarking on broader quality of care indicators. Most surveyed hospitals had implemented some measures to address the NFID-CDC recommendations; however, hospitals need to do much more to improve antimicrobial use and to increase their efforts to detect, report, and control the spread of antimicrobial resistance. A supportive hospital administration must foster a culture of ongoing support, education, and interdisciplinary work groups focused on this important issue to successfully accomplish these goals.


Telehealth adaptation of the Lidcombe Program of Early Stuttering Intervention: five case studies.
This article describes and reports data-based outcomes of a low-tech telehealth adaptation of the Lidcombe Program of Early Stuttering Intervention. Participants were 5 children with early stuttering, ranging in age from 3;5 (years;months) to 5;7, and their families. All children met the speech criteria for completion of Stage 1 of the Lidcombe Program. Data suggested that the treatment method may be viable and that favorable outcomes may be achievable. Mean posttreatment stuttering rates in everyday speaking situations were available 12 months posttreatment for 4 children. Two children scored a mean percent syllables stuttered of less than 1.0 at that time, and 2 children scored a mean of below 2.0. As occurs often in standard delivery of the Lidcombe Progam, 1 child relapsed after Stage 1, apparently because of parental noncompliance, but this relapse was managed successfully. Follow up data were unavailable for 1 child. For 4 of the 5 cases, the number of consultations required exceeded established benchmarks for standard Lidcombe Program delivery, suggesting that telephone-based telehealth may be a less efficient version of the treatment. The implications of these preliminary data are discussed.


Biology and Genomics of an Historic Therapeutic Escherichia coli Bacteriophage Collection.
We have performed microbiological and genomic characterization of an historic collection of nine bacteriophages, specifically infecting a K1 E. coli O18:K1:H7 ColV+ strain. These phages were isolated from sewage and tested for their efficacy in vivo for the treatment of systemic E. coli infection in a mouse infection model by Smith and Huggins (1982). The aim of the study was to identify common microbiological and genomic characteristics, which co-relate to the performance of these phages in in vivo study. These features will allow an informed selection of phages for use as therapeutic agents. Transmission electron microscopy showed that six of the nine phages were Podoviridae and the remaining three were Siphoviridae. The four best performing phages in vivo belonged to the Podoviridae family. In vitro, these phages exhibited very short latent and rise periods in our study. In agreement with their microbiological profiles, characterization by genome sequencing showed that all six podoviruses belong to the Autographivirinae subfamily. Of these, four were isolates of the same species (99% identity), whereas two had divergent genomes compared to other podoviruses. The Siphoviridae phages, which were moderate to poor performers in vivo, exhibited longer latent and rise periods in vitro. Two of the three siphoviruses were closely related to each other (99% identity), but all can be associated with the Guernseyvirinae subfamily. Genome sequence comparison of both types of phages showed that a gene encoding for DNA-dependent RNA polymerase was only present in phages with faster replication cycle, which may account for their better performance in vivo. These data define a combination of microbiological, genomic and in vivo characteristics which allow a more rational evaluation of the original in vivo data and pave the way for the selection of phages for future phage therapy trails.


Burden of comorbidity in individuals with asthma.
Asthma comorbidity, such as depression and obesity, has been associated with greater healthcare use, decreased quality of life and poor asthma control. Treating this comorbidity has been shown to improve asthma outcomes as well as overall health. Despite this, asthma comorbidity remains relatively under-recognised and understudied-perhaps because most asthma occurs in young people who are believed to be healthy and relatively free of comorbidity. The aim of this study was to quantify empirically the amount of comorbidity associated with asthma. A population-based cohort study was conducted using the health administrative data of the 12 million residents of Ontario, Canada in 2005. A validated health administrative algorithm was used to identify individuals with asthma. The amount of comorbidity among individuals with asthma, as reflected in rates of hospitalisations, emergency department visits and ambulatory care claims, was found to be substantial and much greater than that observed among individuals without asthma. Together, asthma and asthma comorbidity (the extra comorbidity found in individuals with asthma compared with those without asthma) were associated with 6% of all hospitalisations, 9% of all emergency room visits and 6% of all ambulatory care visits that occurred in Ontario. Asthma comorbidity places a significant burden on individuals and the healthcare system and should be considered in the management of asthma. Further research should focus on which types of asthma comorbidity are responsible for the greatest burden and how such comorbidity should be prevented and managed.


Changes in nuclear protein acetylation in u.v.-damaged human cells.
We have investigated the levels of nuclear protein acetylation in u.v.-irradiated human fibroblasts. Initially, we measured the levels of acetylation in total acid-soluble nuclear proteins and observed two distinct differences between the irradiated and unirradiated (control) cells. Immediately after irradiation, there is a 'wave' of protein hyperacetylation (i.e. a total acetylation level greater than that of unirradiated cells) that lasts for 2-6 h depending on the experimental conditions. This hyperacetylation phase is then followed by a hypoacetylation phase, lasting for many hours, and the total level of acetylation does not return to that of control cells until 24-72 h after u.v. damage. Both the magnitude and duration of each phase is dependent on the dose of u.v. light used. The wave of hyperacetylation is more pronounced at low u.v. doses (i.e. less than 5 J/m2), while the wave of hypoacetylation is more pronounced at higher u.v. doses (greater than or equal to 8 J/m2). Furthermore, the duration of each phase is prolonged when cells are exposed to 2 mM hydroxyurea, an agent which retards the rate of excision repair at u.v.-damaged sites. Examination of the acetylation levels of the individual nuclear proteins indicated that acetylation of the core histones follows the same pattern observed for the total acid-soluble protein fractions. Furthermore, these were the only major proteins in the total acid-soluble fraction observed to undergo the early, rapid hyperacetylation immediately following u.v. damage. Acetylation of histone H1 was negligible in both damaged and control cells, while three prominent non-histone proteins were acetylated only after long labeling times (greater than 4 h) in each case, gradually becoming hyperacetylated in the u.v.-damaged cells. These results raise the possibility that a causal relationship exists between nuclear protein acetylation and nucleotide excision repair of DNA in human cells.


Binaural Loudness Constancy.
In binaural loudness summation, diotic presentation of a sound usually produces greater loudness than monaural presentation. However, experiments using loudspeaker presentation with and without earplugs find that magnitude estimates of loudness are little altered by the earplug, suggesting a form of loudness constancy. We explored the significance of controlling stimulation of the second ear using meatal occlusion as opposed to the deactivation of one earphone. We measured the point of subjective loudness equality (PSLE) for monaural vs. binaural presentation using an adaptive technique for both speech and noise. These stimuli were presented in a reverberant room over a loudspeaker to the right of the listener, or over lightweight headphones. Using the headphones, stimuli were either presented dry, or matched to those of the loudspeaker by convolution with impulse responses measured from the loudspeaker to the listener position, using an acoustic manikin. The headphone response was also compensated. Using the loudspeaker, monaural presentation was achieved by instructing the listener to block the left ear with a finger. Near perfect binaural loudness constancy was observed using loudspeaker presentation, while there was a summation effect of 3-6 dB for both headphone conditions. However, only partial constancy was observed when meatal occlusion was simulated. These results suggest that there may be contributions to binaural loudness constancy from residual low frequencies at the occluded ear as well as a cognitive element, which is activated by the knowledge that one ear is occluded.


Preventing sintering of nanoclusters on graphene by radical adsorption.
Metal nanoclusters, supported on inert substrates, exhibiting well-defined shapes and sizes in a broad range of temperatures are a major object of desire in nanotechnology. Here, a technique is presented that improves the thermal stability of monodisperse and crystalline transition metal nanoclusters grown in a regular array on metal-supported graphene. To stabilize the clusters after growth under ultrahigh vacuum the system composed of the aggregates and the graphene/metal interface is exposed to radicals resulting from the dissociation of diatomic gases. As a model system we have used Pt as the metal element for cluster growth and the template consisting of the moiré pattern resulting from the lattice mismatch between graphene and the Ir(111) surface. The study has been performed for deuterium and oxygen radicals, which interact very differently with graphene. Our results reveal that after radical exposure the thermally activated motion of Pt nanoclusters to adjacent moiré cells and the subsequent sintering of neighbor aggregates are avoided, most pronounced for the case of atomic O. For the case of D the limits of the improvement are given by radical desorption, whereas for the case of O they are defined by an interplay between coalescence and graphene etching followed by Pt intercalation, which can be controlled by the amount of exposure. Finally, we determined the mechanism of how radical adsorption improves the thermal stability of the aggregates.


Circulating Levels of Betatrophin and Irisin Are Not Associated with Pancreatic β-Cell Function in Previously Diagnosed Type 2 Diabetes Mellitus Patients.
Betatrophin and irisin are two recently identified hormones which may participate in regulating pancreatic β-cell function. However, the associations of these two hormones with β-cell function remain unclear. The present study aims to demonstrate the associations of circulating betatrophin and irisin levels with β-cell function, assessed by the area under the curve (AUC) of C-peptide, and the possible correlation between these two hormones in previously diagnosed type 2 diabetes mellitus (T2DM) patients. In total, 20 age-, sex-, and body mass index- (BMI-) matched normal glucose tolerance (NGT) subjects and 120 previously diagnosed T2DM patients were included in this study. Partial correlation analysis was used to evaluate the relationships between these two hormones and indexes of β-cell function and insulin resistance. Our results showed that betatrophin levels were significantly elevated, while irisin levels were significantly decreased, in patients with T2DM compared with NGT subjects. However, partial correlation analysis showed that betatrophin levels did not correlate with β-cell function-related variables or insulin resistance-related variables before or after controlling multiple covariates, while irisin correlated positively with insulin sensitivity but is not associated with β-cell function-related variables. Besides, no correlation was observed between betatrophin and irisin levels. Hence we concluded that betatrophin and irisin were not associated with β-cell function in previously diagnosed T2DM patients.


Nucleotide-resolution profiling of RNA recombination in the encapsidated genome of a eukaryotic RNA virus by next-generation sequencing.
Next-generation sequencing has been used in numerous investigations to characterize and quantify the genetic diversity of a virus sample through the mapping of polymorphisms and measurement of mutation frequencies. Next-generation sequencing has also been employed to identify recombination events occurring within the genomes of higher organisms, for example, detecting alternative RNA splicing events and oncogenic chromosomal rearrangements. Here, we combine these two approaches to profile RNA recombination within the encapsidated genome of a eukaryotic RNA virus, flock house virus. We detect hundreds of thousands of recombination events, with single-nucleotide resolution, which result in diversity in the encapsidated genome rivaling that due to mismatch mutation. We detect previously identified defective RNAs as well as many other abundant and novel defective RNAs. Our approach is exceptionally sensitive and unbiased and requires no prior knowledge beyond the virus genome sequence. RNA recombination is a powerful driving force behind the evolution and adaptation of RNA viruses. The strategy implemented here is widely applicable and provides a highly detailed description of the complex mutational landscape of the transmissible viral genome.


Acquisition of direct antiviral effector functions by CMV-specific CD4+ T lymphocytes with cellular maturation.
The role of CD4+ T cells in the control of persistent viral infections beyond the provision of cognate help remains unclear. We used polychromatic flow cytometry to evaluate the production of the cytokines interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-2, the chemokine macrophage inflammatory protein (MIP)-1beta, and surface mobilization of the degranulation marker CD107a by CD4+ T cells in response to stimulation with cytomegalovirus (CMV)-specific major histocompatibility complex class II peptide epitopes. Surface expression of CD45RO, CD27, and CD57 on responding cells was used to classify CD4+ T cell maturation. The functional profile of virus-specific CD4+ T cells in chronic CMV infection was unique compared with that observed in other viral infections. Salient features of this profile were: (a) the simultaneous production of MIP-1beta, TNF-alpha, and IFN-gamma in the absence of IL-2; and (b) direct cytolytic activity associated with surface mobilization of CD107a and intracellular expression of perforin and granzymes. This polyfunctional profile was associated with a terminally differentiated phenotype that was not characterized by a distinct clonotypic composition. Thus, mature CMV-specific CD4+ T cells exhibit distinct functional properties reminiscent of antiviral CD8+ T lymphocytes.


Clinical Potential of lncRNA PPP1R26-AS1 in Breast Cancer and Its Contribution to Cancer Progression.
Breast cancer has become the most leading diagnosed tumor worldwide in 2020. In this study, the biomarker potential and influence on the cellular function of lncRNA PPP1R26-AS1 was investigated in breast cancer. Expression levels of lncRNA PPP1R26-AS1 in breast tissues and cells were detected by RT-qPCR. Association between lncRNA PPP1R26-AS1 level and clinical parameters was investigated by Chi-square analysis. The prognostic potential was assessed by Kaplan-Meier and multivariate Cox regression analysis. Knockdown of lncRNA PPP1R26-AS1 was subjected to study the effect on cell proliferation, invasion, and migration by CCK-8 and transwell assay. The bind between PPP1R26-AS1 and miR-1226-3p was investigated. LncRNA PPP1R26-AS1 was highly expressed in breast tissues and cell lines. This upregulation was correlated with pTNM, positive ER status, luminal B subtype, positive nodal status, and shorter overall survival in breast cancer patients. Significant decreases in proliferation, migration, and invasion activity were observed upon knockdown of lncRNA PPP1R26-AS1. lncRNA PPP1R26-AS1 could act as ceRNA to bind with miR-1226-3p in breast cancer. LncRNA PPP1R26-AS1, as oncogenic lncRNA, could provide a new perspective on the development of prognostic biomarkers and a new approach in tailoring the treatment personalized in breast cancer.


Stage IV breast cancer in the era of targeted therapy: does surgery of the primary tumor matter?
Multiple studies have suggested that resection of the primary tumor improves survival in patients with stage IV breast cancer, yet in the era of targeted therapy, the relation between surgery and tumor molecular subtype is unknown. The objective of the current study was to identify subsets of patients who may benefit from primary tumor treatment and assess the frequency of local disease progression. Patients presenting with stage IV breast cancer and intact primary tumors (n = 186) were identified from a prospectively maintained clinical database (2000-2004) and clinical data were abstracted (grading determined according to the American Joint Committee on Cancer staging system). Surgery was performed in 69 (37%) patients: 34 (49%) patients with unknown metastatic disease at the time of surgery, 15 (22%) patients for local control, 14 (20%) patients for palliation, and in 6 (9%) patients to obtain tissue. Surgical patients were more likely to be HER-2/neu negative (P = .001), and to have smaller tumors (P = .05) and solitary metastasis (P <.001). Local therapy included axillary lymph node clearance in 33 (48%) patients and postoperative radiotherapy in 9 (13%) patients. The median survival was 35 months. Cox regression analysis identified estrogen receptor (ER) positivity (hazard ratio [HR], 0.47; 95% confidence interval [95% CI], 0.29-0.76), progesterone receptor (PR) positivity (HR, 0.57; 95% CI, 0.36-0.90), and HER-2/neu amplification (HR, 0.51; 95% CI, 0.34-0.77) as being predictive of improved survival. There was a trend toward improved survival with surgery (HR, 0.71; 95% CI, 0.47-1.06). On exploratory analyses, surgery was found to be associated with improved survival in patients with ER/PR positive or HER-2/neu-amplified disease (P = .004). No survival benefit was observed in patients with triple-negative disease. Although a trend toward improved survival with surgery was observed, it was noted most strongly in patients with ER/PR positive and/or HER-2/neu-amplified disease. This suggests that the impact of local control is greatest in the presence of effective targeted therapy, and supports the need for further study to define patient subsets that will benefit most.


Accuracy of forensic pathologists in incorporating post-mortem CT (PMCT) in forensic death investigation.
Post-mortem computed tomography (PMCT) is now performed routinely in some medical examiner's offices, and the images are typically interpreted by forensic pathologists. In this study, the question of whether pathologists appropriately identify significant PMCT findings and incorporate them into the death investigation report and the cause and manner of death (COD and MOD) statements was addressed. We retrospectively reviewed 200 cases where PMCT was performed. The cases were divided into four categories: (1) full autopsy without radiology consultation (n = 77), (2) external exam without radiology consultation (n = 79), (3) full autopsy with radiology consultation (n = 26), (4) external exam with radiology consultation (n = 18). A radiologist (not the consult radiologist) read the PMCT images, and a pathologist (not the case pathologist) reviewed the case pathologist's post-mortem examination report in tandem to determine any PMCT findings omitted from the report. Omitted findings were classified into error types according to a modified Goldman classification including Major 1: Unrecognized fatal injury or pathology that would change COD and/or MOD, and Major 2: Unrecognized fatal injury or pathology that would not change COD and/or MOD. A total of 13 Major errors were identified (6.5%), and none definitively changed the MOD. All four Major-1 errors which could change the COD were found in Category 2. Of 9 Major-2 errors, 2 occurred in Category 1, 6 occurred in Category 2, and 1 occurred in Category 4. In conclusion, forensic pathologists who routinely utilize computed tomography (CT) interpret CT images well enough to reliably certify the COD and MOD.


Impact of Genetic Determinants of HbA1c on Type 2 Diabetes Risk and Diagnosis.
Glycated hemoglobin (A1c) is used to diagnose type 2 diabetes and monitor glycemic control. Specific genetic variants interfere with A1c and effects/frequencies of some variants vary by ancestry. In this review, we summarize findings from large trans-ethnic meta-analyses of genome-wide association studies (GWAS) of A1c and describe some variants influencing erythrocyte biology and interfering with A1c. Recent GWAS meta-analyses have revealed 60 loci associated with A1c in multi-ethnic populations. The main A1c genetic driver in African Americans is rs1050828 (G6PD). Some identified loci are located in/near genes known as monogenic causes of erythrocytic disorders (ANK1, SPTA1) or iron disorders (TMPRSS6, HFE). Uncommon genetic variants (not revealed by GWAS) that are known to cause hemoglobinopathies may also influence A1C levels, partly by interfering with laboratory assays. Specific genetic variants that have a large impact on A1c levels may influence clinical practice, especially in individuals of African descent. Efforts to reveal novel A1c loci should focus on increasing representation of GWAS in non-European ancestries, and on using better genome-wide coverage of uncommon variants that are specific to each population.


Expression and processing of the AIDS virus reverse transcriptase in Escherichia coli.
The ability to express the genes of pathogenic human viruses, such as the acquired immune deficiency syndrome (AIDS) virus (also called human immunodeficiency virus) in bacterial cells affords the opportunity to study proteins that are ordinarily difficult or inconvenient to obtain in amounts sufficient for detailed analysis. A segment of the AIDS virus pol gene was expressed in Escherichia coli. Expression resulted in the appearance of reverse transcriptase activity in the bacterial cell extracts. The extracts contained two virus-related polypeptides that have the same apparent molecular weights as the two processed forms of virion-derived reverse transcriptase (p66 and p51). The formation of these two polypeptides depended on the coexpression of sequences located near the 5' end of the pol gene, a region that is thought to encode a viral protease. This bacterial system appears to generate mature forms of the AIDS virus reverse transcriptase by a proteolytic pathway equivalent to that which occurs during virus infection of human cells.


Dietary fat and fatty acid profile are associated with indices of skeletal muscle mass in women aged 18-79 years.
Age-related loss of skeletal muscle mass results in a reduction in metabolically active tissue and has been related to the onset of obesity and sarcopenia. Although the causes of muscle loss are poorly understood, dietary fat has been postulated to have a role in determining protein turnover through an influence on both inflammation and insulin resistance. This study was designed to investigate the cross-sectional relation between dietary fat intake, as dietary percentage of fat energy (PFE) and fatty acid profile, with indices of skeletal muscle mass in the population setting. Body composition [fat-free mass (FFM; in kg)] and the fat-free mass index (FFMI; kg FFM/m(2)) was measured by using dual-energy X-ray absorptiometry in 2689 women aged 18-79 y from the TwinsUK Study and calculated according to quintile of dietary fat (by food-frequency questionnaire) after multivariate adjustment. Positive associations were found between the polyunsaturated-to-saturated fatty acid (SFA) ratio and indices of FFM, and inverse associations were found with PFE, SFAs, monounsaturated fatty acids (MUFAs), and trans fatty acids (TFAs) (all as % of energy). Extreme quintile dietary differences for PFE were -0.6 kg for FFM and -0.28 kg/m(2) for FFMI; for SFAs, MUFAs, and TFAs, these were -0.5 to -0.8 kg for FFM and -0.26 to -0.38 kg/m(2) for FFMI. These associations were of a similar magnitude to the expected decline in muscle mass that occurs over 10 y. To our knowledge, this is the first population-based study to demonstrate an association between a comprehensive range of dietary fat intake and FFM. These findings indicate that a dietary fat profile already associated with cardiovascular disease protection may also be beneficial for conservation of skeletal muscle mass.


Intrathoracic giant solitary fibrous tumor.
Solitary fibrous tumor of the pleura is a rare, usually benign, and slow-growing neoplasm. Complete surgical resection for giant tumor of the pleura is challenging because of poor exposure and a large blood supply. We report the case of a giant hypervascular fibrous tumor that filled nearly the entire left hemithorax and anterior mediastinum, and its preoperative management. A 59-year-old woman presented to us with exertional dyspnea and chest pain. A chest radiograph showed the right hemithorax completely opaque and a mediastinal shift to the left hemithorax. A tomography scan of the thorax showed a giant mass that almost completely filled the right hemithorax and compressed the mediastinum to the left. Because of excessive bleeding during dissection, the operation was terminated after a biopsy specimen was obtained. The biopsy was diagnosed as a benign fibrous tumour. A thoracic computed tomography angiogram showed that the mass was supplied by multiple intercostal arteries as well as an aberrant artery that branches off the celiac trunk in the subdiaphragmatic region. Due to the many arteries that needed to be embolized, the final decision was to control the bleeding following resection by inducing total circulatory arrest with the help of cardiopulmonary bypass. The bleeding could not be controlled under cardiopulmonary bypass and the patient's death was confirmed. We report this case to emphasize the necessity of preoperative embolization; the use of cardiopulmonary bypass and total circulatory arrest is not a valid alternative method to control the bleeding.


Rivaroxaban in Rheumatic Heart Disease-Associated Atrial Fibrillation.
Testing of factor Xa inhibitors for the prevention of cardiovascular events in patients with rheumatic heart disease-associated atrial fibrillation has been limited. We enrolled patients with atrial fibrillation and echocardiographically documented rheumatic heart disease who had any of the following: a CHA2DS2VASc score of at least 2 (on a scale from 0 to 9, with higher scores indicating a higher risk of stroke), a mitral-valve area of no more than 2 cm2, left atrial spontaneous echo contrast, or left atrial thrombus. Patients were randomly assigned to receive standard doses of rivaroxaban or dose-adjusted vitamin K antagonist. The primary efficacy outcome was a composite of stroke, systemic embolism, myocardial infarction, or death from vascular (cardiac or noncardiac) or unknown causes. We hypothesized that rivaroxaban therapy would be noninferior to vitamin K antagonist therapy. The primary safety outcome was major bleeding according to the International Society of Thrombosis and Hemostasis. Of 4565 enrolled patients, 4531 were included in the final analysis. The mean age of the patients was 50.5 years, and 72.3% were women. Permanent discontinuation of trial medication was more common with rivaroxaban than with vitamin K antagonist therapy at all visits. In the intention-to-treat analysis, 560 patients in the rivaroxaban group and 446 in the vitamin K antagonist group had a primary-outcome event. Survival curves were nonproportional. The restricted mean survival time was 1599 days in the rivaroxaban group and 1675 days in the vitamin K antagonist group (difference, -76 days; 95% confidence interval [CI], -121 to -31; P<0.001). A higher incidence of death occurred in the rivaroxaban group than in the vitamin K antagonist group (restricted mean survival time, 1608 days vs. 1680 days; difference, -72 days; 95% CI, -117 to -28). No significant between-group difference in the rate of major bleeding was noted. Among patients with rheumatic heart disease-associated atrial fibrillation, vitamin K antagonist therapy led to a lower rate of a composite of cardiovascular events or death than rivaroxaban therapy, without a higher rate of bleeding. (Funded by Bayer; INVICTUS ClinicalTrials.gov number, NCT02832544.).


Running economy and effort after cycling: Effect of methodological choices.
Prior exercise can negatively affect movement economy of a subsequent task. However, the impact of cycling exercise on the energy cost of subsequent running is difficult to ascertain, possibly because of the use of different methods of calculating economy. We examined the influence of a simulated cycling bout on running physiological cost (running economy, heart rate and ventilation rates) and perceptual responses (ratings of perceived exertion and effort) by comparing two running bouts, performed before and after cycling using different running economy calculation methods. Seventeen competitive male triathletes ran at race pace before and after a simulated Olympic-distance cycling bout. Running economy was calculated as V̇O2 (mL∙kg-1∙min-1), oxygen cost (EO2, mL∙kg-1∙m-1) and aerobic energy cost (Eaer, J∙kg-1∙m-1). All measures of running economy and perceptual responses indicated significant alterations imposed by prior cycling. Despite a good level of agreement with minimal bias between calculation methods, differences (p < 0.05) were observed between Eaer and both V̇O2 and EO2. The results confirmed that prior cycling increased physiological cost and perceptual responses in a subsequent running bout. It is recommended that Eaer be calculated as a more valid measure of running economy alongside perceptual responses to assist in the identification of individual responses in running economy following cycling.


Comparative diagnostic accuracy of serum levels of neutrophil activating peptide-2 and pentraxin-3 versus troponin-I in acute coronary syndrome.
We measured the levels of neutrophil activating peptide-2 (NAP-2) and pentraxin-3 (PTX-3) in acute coronary syndromes (ACS) patients and compared their diagnostic accuracy with cardiac troponin I (cTnI). We conducted a prospective cohort study to determine the diagnostic accuracy of PTX-3, NAP-2 and cTnI for the prediction of ACS. Consecutively eighty-three patients with sudden chest pain admitted to Dicle University Emergency Department within the first six hours of symptom onset were included in our study. Mean serum levels of PTX-3, NAP-2 and cTnI were compared between control and patient groups and ACS subgroups. Their sensitivities and specificities in early diagnosis of ACS were identified. Receiver operating characteristic (ROC) analysis was used to assess the diagnostic validity of the markers, and areas under the ROC curve (AUC) were compared. In the patient group, mean serum concentrations of NAP-2 (53.03+22.77 ng/ml) and PTX-3 (1.73+0.82 ng/ml) were considerably higher than those of the control group (24.54+9.50 and 0.50+0.39 ng/ml, respectively) (p<0.01). When compared with the control group, PTX-3 levels of all three ACS subtypes (unstable angina pectoris (USAP) - 1.62+0.41 ng/ml, non-ST elevation myocardial infarction (NSTEMI) -1.63+0.31 ng/ml and ST-elevation myocardial infarction (STEMI) - 1.75+0.89 ng/ml) were higher, whereas NAP-2 levels were higher in USAP (56.29+22.60 ng/ml) and STEMI (52.05+20.99 ng/ml) patients (p<0.01). For diagnosing ACS within the first six hours of presentation, PTX-3 sensitivity was 98.5% and specificity was 92.3%, and NAP-2 sensitivity - 98.1% and specificity - 41.3%. The ROC curve AUC values were: 0.962 for PTX-3 (95% CI 0.802 - 1.073), 0.840 for NAP-2 (95% CI 0.684 - 0.991), and 0.683 for cTnI (95% CI 0.610 - 0.940). Pentraxin-3 is a sensitive and specific marker for ACS diagnosis when compared with cardiac markers in patients admitted to the emergency department (ED) within the first six hours of onset of chest pain.


Differential Effects of Influenza Virus NA, HA Head, and HA Stalk Antibodies on Peripheral Blood Leukocyte Gene Expression during Human Infection.
In this study, we examined the relationships between anti-influenza virus serum antibody titers, clinical disease, and peripheral blood leukocyte (PBL) global gene expression during presymptomatic, acute, and convalescent illness in 83 participants infected with 2009 pandemic H1N1 virus in a human influenza challenge model. Using traditional statistical and logistic regression modeling approaches, profiles of differentially expressed genes that correlated with active viral shedding, predicted length of viral shedding, and predicted illness severity were identified. These analyses further demonstrated that challenge participants fell into three peripheral blood leukocyte gene expression phenotypes that significantly correlated with different clinical outcomes and prechallenge serum titers of antibodies specific for the viral neuraminidase, hemagglutinin head, and hemagglutinin stalk. Higher prechallenge serum antibody titers were inversely correlated with leukocyte responsiveness in participants with active disease and could mask expression of peripheral blood markers of clinical disease in some participants, including viral shedding and symptom severity. Consequently, preexisting anti-influenza antibodies may modulate PBL gene expression, and this must be taken into consideration in the development and interpretation of peripheral blood diagnostic and prognostic assays of influenza infection.IMPORTANCE Influenza A viruses are significant human pathogens that caused 83,000 deaths in the United States during 2017 to 2018, and there is need to understand the molecular correlates of illness and to identify prognostic markers of viral infection, symptom severity, and disease course. Preexisting antibodies against viral neuraminidase (NA) and hemagglutinin (HA) proteins play a critical role in lessening disease severity. We performed global gene expression profiling of peripheral blood leukocytes collected during acute and convalescent phases from a large cohort of people infected with A/H1N1pdm virus. Using statistical and machine-learning approaches, populations of genes were identified early in infection that correlated with active viral shedding, predicted length of shedding, or disease severity. Finally, these gene expression responses were differentially affected by increased levels of preexisting influenza antibodies, which could mask detection of these markers of contagiousness and disease severity in people with active clinical disease.


An [18F]dopa-PET and clinical study of the rate of progression in Parkinson's disease.
We have studied disease progression in a group of 10 patients with recent onset idiopathic Parkinson's disease [mean age 53.7 +/- 12.6 years, mean duration 18.3 +/- 6.5 months, mean unified Parkinson's disease rating scale (UPDRS) 19.0 +/- 9.5], and a group of seven patients with established Parkinson's disease (mean age 60.1 +/- 15.1 years, mean duration 70.8 +/- 35.5 months, mean UPDRS 47.9 +/- 18.1), using both clinical assessment and [18F]dopa-PET. Results were compared with those of a group of 10 normal subjects (age 66 +/- 16 years). Mean putamen [18F]dopa influx constant (Ki) was a reliable measurement (R = 0.82) and the most sensitive marker of disease progression (r = -0.85, P < 0.0001). The mean annual rate of reduction in mean putamen Ki in the Parkinson's disease patients was 12.5% per annum, whereas the control group showed no significant change in Ki over a mean of 32 months follow up. The rate of progression was more rapid in the recent onset compared with the established disease group but this did not reach statistical significance. Assuming a linear progression for the entire group we estimate symptom onset at a mean putamen Ki 79% of normal with a mean preclinical period of 3.1 years.


Analysis of genetic variations of protein tyrosine kinase fyn and their association with alcohol dependence in two independent cohorts.
Decreased sensitivity to and increased tolerance for the effects of alcohol is a phenotype, which was shown to be associated with an increased risk for alcoholism in humans and was observed in protein tyrosine kinase (PTK) fyn knockout mice. We performed an association study of genetic variations of PTK fyn in 430 alcohol-dependent patients and 365 unrelated control subjects from two independent samples. In a combined analysis, we found an association of alcohol dependence with the single nucleotide polymorphism (SNP) T137346C in the 5' untranslated region (UTR) of the gene. A relevant association could be excluded for the remaining two informative SNPs. Selection by phenotype showed that a high number of withdrawal symptoms, high amount of alcohol intake, and high maximum number of drinks compared with unrelated control subjects was associated with the SNP in the 5'-UTR region but not with the remaining SNPs. Our results indicate a possible association of alcohol dependence with a genotype of the SNP T137346C of the PTK fyn, with C being the risk allele.


Magnetic micro/nanoparticle flocculation-based signal amplification for biosensing.
We report a time and cost efficient signal amplification method for biosensors employing magnetic particles. In this method, magnetic particles in an applied external magnetic field form magnetic dipoles, interact with each other, and accumulate along the magnetic field lines. This magnetic interaction does not need any biomolecular coating for binding and can be controlled with the strength of the applied magnetic field. The accumulation can be used to amplify the corresponding pixel area that is obtained from an image of a single magnetic particle. An application of the method to the Escherichia coli 0157:H7 bacteria samples is demonstrated in order to show the potential of the approach. A minimum of threefold to a maximum of 60-fold amplification is reached from a single bacteria cell under a magnetic field of 20 mT.


The oleocanthal-based homovanillyl sinapate as a novel c-Met inhibitor.
The hepatocyte growth factor (HGF)/mesenchymal-epithelial transition factor (c-Met) signaling axis has gained considerable attention as an attractive molecular target for therapeutic blockade of cancer. Inspired by the chemical structure of S (-)-oleocanthal, a natural secoiridoid from extra-virgin olive oil with documented anticancer activity against c-Met-dependent malignancies, the research presented herein reports on the discovery of the novel olive-derived homovanillyl sinapate (HVS) as a promising c-Met inhibitor. HVS was distinguished for its remarkable potency against wild-type c-Met and its oncogenic variant in cell-free assays and confirmed by in silico docking studies. Furthermore, HVS substantially impaired the c-Met-mediated growth across a broad spectrum of breast cancer cells, while similar treatment doses had no effect on the non-tumorigenic mammary epithelial cell growth. In addition, HVS caused a dose-dependent inhibition of HGF-induced, but not epidermal growth factor (EGF)-induced, cell scattering in addition to HGF-mediated migration, invasion, and 3-dimensional (3D) proliferation of tumor cell spheroids. HVS treatment effects were mediated via inhibition of ligand-mediated c-Met activation and its downstream mitogenic signaling and blocking molecular mediators involved in cellular motility across different cellular contexts. An interesting feature of HVS is its good selectivity for c-Met and Abelson murine leukemia viral oncogene homolog 1 (ABL1) when profiled against a panel of kinases. Docking studies revealed interactions likely to impart high dual affinity for both ABL1 and c-Met kinases. HVS markedly reduced tumor growth, showed excellent pharmacodynamics, and suppressed cell proliferation and microvessel density in an orthotopic model of triple negative breast cancer. Collectively, the present findings suggested that the oleocanthal-based HVS is a promising c-Met inhibitor lead entity with excellent therapeutic potential to control malignancies with aberrant c-Met activity.


Acute thrombosis of the splanchnic veins.
The consequence of an acute thrombosis in the splanchnic veins on the viability of the intestine has not been well defined in the literature. Spontaneous recovery or total necrosis of the bowel have both been described. We treated seven patients with thrombosis of the splanchnic veins and adopted a surgical approach in three patients with extended and complete thrombosis of the superior mesenteric vein, portal vein, and splenic vein, while four patients with partial thrombosis of the superior mesenteric vein or protal vein recovered with conservative treatment. A 22-year literature review has identified 64 cases of acute thrombosis in the splanchnic veins, with complete information regarding the location and extent of the thrombosis, the treatment, and the outcome. Different anatomical patterns of thrombosis with mortality rates varying between 0% and 76% seem to be related to the extent and completeness of venous obstruction.


The mammalian Tolloid-like 1 gene, Tll1, is necessary for normal septation and positioning of the heart.
Mammalian Tolloid-like 1 (mTLL-1) is an astacin-like metalloprotease, highly similar in domain structure to the morphogenetically important proteases bone morphogenetic protein-1 (BMP-1) and Drosophila Tolloid. To investigate possible roles for mTLL-1 in mammalian development, we have used gene targeting in ES cells to produce mice with a disrupted allele for the corresponding gene, Tll1. Homozygous mutants were embryonic lethal, with death at mid-gestation from cardiac failure and a unique constellation of developmental defects that were apparently confined solely to the heart. Constant features were incomplete formation of the muscular interventricular septum and an abnormal and novel positioning of the heart and aorta. Consistent with roles in cardiac development, Tll1 expression was specific to precardiac tissue and endocardium in 7.5 and 8.5 days p.c. embryos, respectively. Tll1 expression was also high in the developing interventricular septum, where expression of the BMP-1 gene, Bmp1, was not observed. Cardiac structures that were not affected in Tll1-/- embryos either showed no Tll1 expression (atrio-ventricular cushions) or showed overlapping expression of Tll1 and Bmp1 (aortico-pulmonary septum), suggesting that products of the Bmp1 gene may be capable of functionally substituting for mTLL-1 at sites in which they are co-expressed. Together, the various data show that mTLL-1 plays multiple roles in formation of the mammalian heart and is essential for formation of the interventricular septum.


Detection of Theileria parva in the salivary glands of Rhipicephalus appendiculatus: evaluation of staining methods.
A comparison of ten methods for staining tick salivary glands for detection of Theileria parva infection from ticks fed on rabbits for various periods was undertaken. Staining with azure without hydrochloric acid hydrolysis was found to be the most reliable method for detection of the presporozoite stages (sporoblasts) of T. parva in the salivary gland acini of unfed Rhipicephalus appendiculatus and could be applied to field ticks. All the stains proved suitable for the detection and quantitation of sporozoites in ticks fed for 4 days on rabbits. The capacity of the stains to allow detection of early stages of T. parva differed, but it became more reliable during tick feeding as sporoblasts developed and matured. Giemsa's stain and Feulgen's stain followed by superimposition of Giemsa's stain were superior to other stains for the detection and quantitation of immature salivary gland stages in feeding ticks.


Day-care attendance, position in sibship, and early childhood wheezing: a population-based birth cohort study.
There are conflicting data on the effect of day-care attendance and position in sibship on the development of wheezing. To investigate the relationship between day-care attendance and position in sibship with early childhood wheeze. Prospective population-based birth cohort. At age 5 years, we collected information on parentally reported symptoms (n = 922); lung function was ascertained using plethysmography (n = 745) and allergic sensitization by skin testing (n = 815). Participants were assigned into categories according to the age of entry to day-care (0-6, 6-12, >12 mo) and number of older siblings (0, 1, 2, >2). Current wheeze was reported by 203 participants (22%); 224 (28%) were sensitized. In the multivariate model, sensitization (odds ratio, 2.47; 95% CI, 1.66-3.67), male sex (1.49, 1.01-2.20), maternal asthma (1.72, 1.10-2.68), and maternal smoking during pregnancy (2.15, 1.26-3.66) significantly increased the risk of wheezing. Entering day-care between 6 and 12 or after 12 months of age was significantly and inversely associated with current wheeze (0.25, 0.11-0.60; and 0.65, 0.44-0.98, respectively). Entry into nursery between 6 and 12 months reduced the risk of persistent wheezing (P = .04). We found no association between day-care attendance and lung function. Entering nursery in the first 6 months of life was associated with increased risk of atopy (2.47, 1.23-4.95). Having older siblings was associated only with rhinoconjunctivitis (0.72, 0.54-0.97). Day-care attendance was associated with a reduced risk of current wheezing in 5-year-old children. The protective effect appeared strongest for children who entered day-care between the ages of 6 and 12 months.


Nanoparticle-stabilized microbubbles for multimodal imaging and drug delivery.
Microbubbles (MBs) are routinely used as contrast agents for ultrasound imaging. The use of ultrasound in combination with MBs has also attracted attention as a method to enhance drug delivery. We have developed a technology platform incorporating multiple functionalities, including imaging and therapy in a single system consisting of MBs stabilized by polyethylene glycol (PEG)-coated polymeric nanoparticles (NPs). The NPs, containing lipophilic drugs and/or contrast agents, are composed of the widely used poly(butyl cyanoacrylate) (PBCA) polymer and prepared in a single step. MBs stabilized by these NPs are subsequently prepared by self-assembly of NPs at the MB air-liquid interface. Here we show that these MBs can act as contrast agents for conventional ultrasound imaging. Successful encapsulation of iron oxide NPs inside the PBCA NPs is demonstrated, potentially enabling the NP-MBs to be used as magnetic resonance imaging (MRI) and/or molecular ultrasound imaging contrast agents. By precise tuning of the applied ultrasound pulse, the MBs burst and the NPs constituting the shell are released. This could result in increased local deposit of NPs into target tissue, providing improved therapy and imaging contrast compared with freely distributed NPs.


Characteristics, sexual behaviour and risk factors of female, male and transgender sex workers in South Africa.
In South Africa, information on sex workers' characteristics, sexual behaviour and health needs is limited. Current social, legal and institutional factors impede a safe working environment for sex workers and their clients. To describe characteristics and sexual behaviour of female, male and transgender sex workers, and assess their risk factors for unprotected sex. Repeat cross-sectional surveys among sex workers were conducted in Hillbrow, Sandton, Rustenburg and Cape Town in 2010. Sex workers were interviewed once; any re-interviews were excluded from analysis. Unprotected sex was defined as any unprotected penetrative vaginal or anal sex with last two clients. Trained sex workers interviewed 1 799 sex workers. Sex work was a full-time profession for most participants. About 8% (126/1 594) of women, 33% (22/75) of men, and 25% (12/50) of transgender people had unprotected sex. A quarter of anal sex was unprotected. Unprotected sex was 2.1 times (adjusted odds ratio (AOR), 95% CI 1.2 - 3.7; p=0.011) more likely in participants reporting daily or weekly binge drinking than non-binge drinkers. Male sex workers were 2.9 times (AOR, 95%CI 1.6 - 5.3; p<0.001) more likely, and transgender people 2.4 times (AOR, 95% CI 1.1 - 4.9; p=0.021) more likely, than females to have unprotected sex. Sex workers in Hillbrow, where the only sex work-specific clinic was operational, were less likely to have unprotected sex than those in other sites. Tailored sex work interventions should explicitly include male and transgender sex workers, sex work-specific clinics, focus on the risks of unprotected anal sex, and include interventions to reduce alcohol harm.


Co-infection by alveolate parasites and frog virus 3-like ranavirus during an amphibian larval mortality event in Florida, USA.
A multispecies amphibian larval mortality event, primarily affecting American bullfrogs Lithobates catesbeianus, was investigated during April 2011 at the Mike Roess Gold Head Branch State Park, Clay County, Florida, USA. Freshly dead and moribund tadpoles had hemorrhagic lesions around the vent and on the ventral body surface, with some exhibiting a swollen abdomen. Bullfrogs (100%), southern leopard frogs L. sphenocephalus (33.3%), and gopher frogs L. capito (100%) were infected by alveolate parasites. The intensity of infection in bullfrog livers was high. Tadpoles were evaluated for frog virus 3 (FV3) by histology and PCR. For those southern leopard frog tadpoles (n = 2) whose livers had not been obscured by alveolate spore infection, neither a pathologic response nor intracytoplasmic inclusions typically associated with clinical infections of FV3-like ranavirus were noted. Sequencing of a portion (496 bp) of the viral major capsid protein gene confirmed FV3-like virus in bullfrogs (n = 1, plus n = 6 pooled) and southern leopard frogs (n = 1, plus n = 4 pooled). In July 2011, young-of-the-year bullfrog tadpoles (n = 7) were negative for alveolate parasites, but 1 gopher frog tadpole was positive. To our knowledge, this is the first confirmed mortality event for amphibians in Florida associated with FV3-like virus, but the extent to which the virus played a primary role is uncertain. Larval mortality was most likely caused by a combination of alveolate parasite infections, FV3-like ranavirus, and undetermined etiological factors.


The bacterial pathogen Listeria monocytogenes: an emerging model in prokaryotic transcriptomics.
A major challenge in bacterial pathogenesis is understanding the molecular basis of the switch from saprophytism to virulence. Following a recent whole-genome transcriptomic analysis using tiling arrays, an article published in BMC Genomics reports the first use of RNA-seq in Listeria monocytogenes in order to identify genes controlled by sigma B, a transcriptional regulator with a critical role in virulence.


Co-Translational Protein Folding and Sorting in Chloroplasts.
Cells depend on the continuous renewal of their proteome composition during the cell cycle and in order to replace aberrant proteins or to react to changing environmental conditions. In higher eukaryotes, protein synthesis is achieved by up to five million ribosomes per cell. With the fast kinetics of translation, the large number of newly made proteins generates a substantial burden for protein homeostasis and requires a highly orchestrated cascade of factors promoting folding, sorting and final maturation. Several of the involved factors directly bind to translating ribosomes for the early processing of emerging nascent polypeptides and the translocation of ribosome nascent chain complexes to target membranes. In plant cells, protein synthesis also occurs in chloroplasts serving the expression of a relatively small set of 60-100 protein-coding genes. However, most of these proteins, together with nucleus-derived subunits, form central complexes majorly involved in the essential processes of photosynthetic light reaction, carbon fixation, metabolism and gene expression. Biogenesis of these heterogenic complexes adds an additional level of complexity for protein biogenesis. In this review, we summarize the current knowledge about co-translationally binding factors in chloroplasts and discuss their role in protein folding and ribosome translocation to thylakoid membranes.


Poorly processed reusable surface disinfection tissue dispensers may be a source of infection.
Reusable surface disinfectant tissue dispensers are used in hospitals in many countries because they allow immediate access to pre-soaked tissues for targeted surface decontamination. On the other hand disinfectant solutions with some active ingredients may get contaminated and cause outbreaks. We determined the frequency of contaminated surface disinfectant solutions in reusable dispensers and the ability of isolates to multiply in different formulations. Reusable tissue dispensers with different surface disinfectants were randomly collected from healthcare facilities. Solutions were investigated for bacterial contamination. The efficacy of two surface disinfectants was determined in suspension tests against two isolated species directly from a contaminated solution or after 5 passages without selection pressure in triplicate. Freshly prepared use solutions were contaminated to determine survival of isolates. 66 dispensers containing disinfectant solutions with surface-active ingredients were collected in 15 healthcare facilities. 28 dispensers from nine healthcare facilities were contaminated with approximately 107 cells per mL of Achromobacter species 3 (9 hospitals), Achromobacter xylosoxidans or Serratia marcescens (1 hospital each). In none of the hospitals dispenser processing had been adequately performed. Isolates regained susceptibility to the disinfectants after five passages without selection pressure but were still able to multiply in different formulations from different manufacturers at room temperature within 7 days. Neglecting adequate processing of surface disinfectant dispensers has contributed to frequent and heavy contamination of use-solutions based on surface active ingredients. Tissue dispenser processing should be taken seriously in clinical practice.


Global burden of acute lower respiratory infection associated with human metapneumovirus in children under 5 years in 2018: a systematic review and modelling study.
Human metapneumovirus is a common virus associated with acute lower respiratory infections (ALRIs) in children. No global burden estimates are available for ALRIs associated with human metapneumovirus in children, and no licensed vaccines or drugs exist for human metapneumovirus infections. We aimed to estimate the age-stratified human metapneumovirus-associated ALRI global incidence, hospital admissions, and mortality burden in children younger than 5 years. We estimated the global burden of human metapneumovirus-associated ALRIs in children younger than 5 years from a systematic review of 119 studies published between Jan 1, 2001, and Dec 31, 2019, and a further 40 high quality unpublished studies. We assessed risk of bias using a modified Newcastle-Ottawa Scale. We estimated incidence, hospital admission rates, and in-hospital case-fatality ratios (hCFRs) of human metapneumovirus-associated ALRI using a generalised linear mixed model. We applied incidence and hospital admission rates of human metapneumovirus-associated ALRI to population estimates to yield the morbidity burden estimates by age bands and World Bank income levels. We also estimated human metapneumovirus-associated ALRI in-hospital deaths and overall human metapneumovirus-associated ALRI deaths (both in-hospital and non-hospital deaths). Additionally, we estimated human metapneumovirus-attributable ALRI cases, hospital admissions, and deaths by combining human metapneumovirus-associated burden estimates and attributable fractions of human metapneumovirus in laboratory-confirmed human metapneumovirus cases and deaths. In 2018, among children younger than 5 years globally, there were an estimated 14·2 million human metapneumovirus-associated ALRI cases (uncertainty range [UR] 10·2 million to 20·1 million), 643 000 human metapneumovirus-associated hospital admissions (UR 425 000 to 977 000), 7700 human metapneumovirus-associated in-hospital deaths (2600 to 48 800), and 16 100 overall (hospital and community) human metapneumovirus-associated ALRI deaths (5700 to 88 000). An estimated 11·1 million ALRI cases (UR 8·0 million to 15·7 million), 502 000 ALRI hospital admissions (UR 332 000 to 762 000), and 11 300 ALRI deaths (4000 to 61 600) could be causally attributed to human metapneumovirus in 2018. Around 58% of the hospital admissions were in infants under 12 months, and 64% of in-hospital deaths occurred in infants younger than 6 months, of which 79% occurred in low-income and lower-middle-income countries. Infants younger than 1 year have disproportionately high risks of severe human metapneumovirus infections across all World Bank income regions and all child mortality settings, similar to respiratory syncytial virus and influenza virus. Infants younger than 6 months in low-income and lower-middle-income countries are at greater risk of death from human metapneumovirus-associated ALRI than older children and those in upper-middle-income and high-income countries. Our mortality estimates demonstrate the importance of intervention strategies for infants across all settings, and warrant continued efforts to improve the outcome of human metapneumovirus-associated ALRI among young infants in low-income and lower-middle-income countries. Bill & Melinda Gates Foundation.


Revisiting sample size: are big trials the answer?
The superiority of the evidence generated in randomized controlled trials over observational data is not only conditional to randomization. Randomized controlled trials require proper design and implementation to provide a reliable effect estimate. Adequate random sequence generation, allocation implementation, analyses based on the intention-to-treat principle, and sufficient power are crucial to the quality of a randomized controlled trial. Power, or the probability of the trial to detect a difference when a real difference between treatments exists, strongly depends on sample size. The quality of orthopaedic randomized controlled trials is frequently threatened by a limited sample size. This paper reviews basic concepts and pitfalls in sample-size estimation and focuses on the importance of large trials in the generation of valid evidence.


Extending peripersonal space representation without tool-use: evidence from a combined behavioral-computational approach.
Stimuli from different sensory modalities occurring on or close to the body are integrated in a multisensory representation of the space surrounding the body, i.e., peripersonal space (PPS). PPS dynamically modifies depending on experience, e.g., it extends after using a tool to reach far objects. However, the neural mechanism underlying PPS plasticity after tool use is largely unknown. Here we use a combined computational-behavioral approach to propose and test a possible mechanism accounting for PPS extension. We first present a neural network model simulating audio-tactile representation in the PPS around one hand. Simulation experiments showed that our model reproduced the main property of PPS neurons, i.e., selective multisensory response for stimuli occurring close to the hand. We used the neural network model to simulate the effects of a tool-use training. In terms of sensory inputs, tool use was conceptualized as a concurrent tactile stimulation from the hand, due to holding the tool, and an auditory stimulation from the far space, due to tool-mediated action. Results showed that after exposure to those inputs, PPS neurons responded also to multisensory stimuli far from the hand. The model thus suggests that synchronous pairing of tactile hand stimulation and auditory stimulation from the far space is sufficient to extend PPS, such as after tool-use. Such prediction was confirmed by a behavioral experiment, where we used an audio-tactile interaction paradigm to measure the boundaries of PPS representation. We found that PPS extended after synchronous tactile-hand stimulation and auditory-far stimulation in a group of healthy volunteers. Control experiments both in simulation and behavioral settings showed that the same amount of tactile and auditory inputs administered out of synchrony did not change PPS representation. We conclude by proposing a simple, biological-plausible model to explain plasticity in PPS representation after tool-use, which is supported by computational and behavioral data.


Rapid growth is a dominant predictor of hepcidin suppression and declining ferritin in Gambian infants.
Iron deficiency and iron deficiency anemia are highly prevalent in low-income countries, especially among young children. Hepcidin is the major regulator of systemic iron homeostasis. It controls dietary iron absorption, dictates whether absorbed iron is made available in circulation for erythropoiesis and other iron-demanding processes, and predicts response to oral iron supplementation. Understanding how hepcidin is itself regulated is therefore important, especially in young children. We investigated how changes in iron-related parameters, inflammation and infection status, seasonality, and growth influenced plasma hepcidin and ferritin concentrations during infancy using longitudinal data from two birth cohorts of infants in rural Gambia (n=114 and n=193). This setting is characterized by extreme seasonality, prevalent childhood anemia, undernutrition, and frequent infection. Plasma was collected from infants at birth and at regular intervals, up to 12 months of age. Hepcidin, ferritin and plasma iron concentrations declined markedly during infancy, with reciprocal increases in soluble transferrin receptor and transferrin concentrations, indicating declining iron stores and increasing tissue iron demand. In cross-sectional analyses at 5 and 12 months of age, we identified expected relationships of hepcidin with iron and inflammatory markers, but also observed significant negative associations between hepcidin and antecedent weight gain. Correspondingly, longitudinal fixed effects modeling demonstrated weight gain to be the most notable dynamic predictor of decreasing hepcidin and ferritin through infancy across both cohorts. Infants who grow rapidly in this setting are at particular risk of depletion of iron stores, but since hepcidin concentrations decrease with weight gain, they may also be the most responsive to oral iron interventions.


Intervention induced changes on parenting practices, youth self-pride and sexual norms to reduce HIV-related behaviors among rural African American youths.
AIDS is the leading killer of African Americans between the ages of 25 and 44, many of whom became infected when they were teenagers or young adults. The disparity in HIV infection rate among African Americans youth residing in rural Southern regions of the United States suggests that there is an urgent need to identify ways to promote early preventive intervention to reduce HIV-related risk behavior. The Strong African American Families (SAAF) program, a preventive intervention for rural African American parents and their 11-year-olds, was specially designed to deter early sexual onset and the initiation and escalation of alcohol and drug use among rural African American preadolescents. A clustered-randomized prevention trial was conducted, contrasting families who took part in SAAF with control families. The trial, which included 332 families, indicated that intervention-induced changes occurred in intervention-targeted parenting, which in turn facilitated changes in youths' internal protective processes and positive sexual norms. Long-term follow up assessments when youth were 17 years old revealed that intervention-induced changes in parenting practices mediated the effect of intervention-group influences on changes in the onset and escalation of risky sexual behaviors over 65 months through its positive influence on adolescents' self-pride and their sexual norms. The findings underscore the powerful effects of parenting practices among rural African American families that over time serve a protective role in reducing youth's risk behavior, including HIV vulnerable behaviors.


Framework for the assessment of interaction between CO2 geological storage and other sedimentary basin resources.
Sedimentary basins around the world considered suitable for carbon storage usually contain other natural resources such as petroleum, coal, geothermal energy and groundwater. Storing carbon dioxide in geological formations in the basins adds to the competition for access to the subsurface and the use of pore space where other resource-based industries also operate. Managing potential impacts that industrial-scale injection of carbon dioxide may have on other resource development must be focused to prevent potential conflicts and enhance synergies where possible. Such a sustainable coexistence of various resource developments can be accomplished by implementing a Framework for Basin Resource Management strategy (FBRM). The FBRM strategy utilizes the concept of an Area of Review (AOR) for guiding development and regulation of CO2 geological storage projects and for assessing their potential impact on other resources. The AOR is determined by the expected physical distribution of the CO2 plume in the subsurface and the modelled extent of reservoir pressure increase resulting from the injection of the CO2. This information is used to define the region to be characterised and monitored for a CO2 injection project. The geological characterisation and risk- and performance-based monitoring will be most comprehensive within the region of the reservoir containing the carbon dioxide plume and should consider geological features and wells continuously above the plume through to its surface projection; this region defines where increases in reservoir pressure will be greatest and where potential for unplanned migration of carbon dioxide is highest. Beyond the expanse of the carbon dioxide plume, geological characterisation and monitoring should focus only on identified features that could be a potential migration conduit for either formation water or carbon dioxide.


Statistical methods for composite endpoints.
Composite endpoints are commonly used in clinical trials, and time-to-first-event analysis has been the usual standard. Time-to-first-event analysis treats all components of the composite endpoint as having equal severity and is heavily influenced by short-term components. Over the last decade, novel statistical approaches have been introduced to overcome these limitations. We reviewed win ratio analysis, competing risk regression, negative binomial regression, Andersen-Gill regression, and weighted composite endpoint (WCE) analysis. Each method has both advantages and limitations. The advantage of win ratio and WCE analyses is that they take event severity into account by assigning weights to each component of the composite endpoint. These weights should be pre-specified because they strongly influence treatment effect estimates. Negative binomial regression and Andersen-Gill analyses consider all events for each patient -rather than only the first event - and tend to have more statistical power than time-to-first-event analysis. Pre-specified novel statistical methods may enhance our understanding of novel therapy when components vary substantially in severity and timing. These methods consider the specific types of patients, drugs, devices, events, and follow-up duration.


Planning impairments in schizophrenia: specificity, task independence and functional relevance.
The present study investigated the specificity of planning impairments in schizophrenia compared to unipolar major depression. Multiple measures of planning ability were employed to assess the task independence of a planning deficit. Furthermore, the predictive power of planning ability with regard to functional outcome was analyzed. A total of 80 participants completed a comprehensive neuropsychological assessment with an emphasis on executive functions and planning ability. The sample consisted of 28 patients with schizophrenia, 28 patients with depression and 24 healthy controls. Both patient groups were impaired on measures of attention, working memory and planning, but only planning ability differentiated between patient groups. The deficit was evident across different measures of planning ability and was the best overall predictor of functional outcome. These results provide evidence for the relative specificity of a planning deficit in schizophrenia and show that the deficit is not task-specific but likely affects central cognitive control processes critical for planned behavior. The observed relation to functional outcome supports the clinical relevance of planning ability.


Declining malaria transmission in rural Amazon: changing epidemiology and challenges to achieve elimination.
In recent years, considerable success in reducing its incidence has been achieved in Brazil, leading to a relative increase in the proportion of cases caused by Plasmodium vivax, considered a harder-to-eliminate parasite. This study aim is to describe the transmission dynamics and associated risk factors in a rural settlement area in the Western Brazilian Amazon. A prospective cohort was established in a rural settlement area for 3 years. Follow-up included continuous passive case detection and monthly active case detection for a period of 6 months. Demographic, clinical and transmission control practices data were collected. Malaria diagnosis was performed through thick blood smear. Univariable and multivariable analyses of factors associated with malaria incidence were performed using negative binomial regression models. Factors associated with recurrence of P. vivax and Plasmodium falciparum malaria within 90 days of a previous episode were analysed using univariable and multivariable Cox-Proportional Hazard models. Malaria prevalence decreased from 7 % at the study beginning to 0.6 % at month 24, with P. vivax predominating and P. falciparum disappearing after 1 year of follow-up. Malaria incidence was significantly higher in the dry season [IRR (95 % CI) 1.4 (1.1-1.6); p < 0.001)]. Use of ITN was associated to malaria protection in the localities [IRR (95 % CI) 0.7 (0.6-0.8); p = 0.001)]. A recurrent P. vivax episode within 90 days was observed in 29.4 % of individuals after an initial diagnosis. A previous P. vivax [IRR (95 % CI) 2.3 (1.3-4.0); p = 0.006)] or mixed P. vivax + P. falciparum [IRR (95 % CI) 2.9 (1.5-5.7); p = 0.002)] infections were significantly associated to a vivax malaria episode within 90 days of follow-up. In an area of P. falciparum and P. vivax co-endemicity, a virtual disappearance of P. falciparum was observed with P. vivax increasing its relative contribution, with a large proportion of recurring episodes. This finding reinforces the perception of P. falciparum being more responsive to early diagnosis and treatment and ITN use and the contribution of relapsing P. vivax to maintain this species' transmission. In areas of P. vivax endemicity, antihypnozoite treatment effectiveness assessment in different transmission intensity may be a fundamental activity for malaria control and elimination.


Reduced satiating effect of d-fenfluramine in serotonin 5-HT(2C) receptor mutant mice.
d-Fenfluramine stimulates the release of serotonin (5-HT) and is a potent inhibitor of the re-uptake of 5-HT into nerve terminals. Administration of d-fenfluramine suppresses food intake in both animals and humans. We have investigated the role of the 5-HT2C receptor in mediating the effect of d-fenfluramine on mouse food intake and the behavioural satiety sequence. Mutant mice lacking serotonin 5-HT2C receptors and wild-type animals were habituated to a daily presentation of wet mash. Animals were non-deprived and received d-fenfluramine (3-30 mg/kg) 30 min prior to being assessed for the presence of stereotypy and presented with wet mash. The behaviour of animals was observed for the subsequent 40 min and food intake was recorded. d-Fenfluramine dose-dependently inhibited the consumption of a palatable wet mash by the mice. d-Fenfluramine (3 mg/kg) significantly reduced the amount of wet mash consumed by wild-type mice and induced a temporal advance in the behavioural satiety sequence consistent with an enhancement of satiety. Mutant mice were less sensitive to the satiating effects of 3 mg/kg d-fenfluramine. Hence, this dose of d-fenfluramine had a reduced effect on both food consumption and the behavioural satiety sequence in the 5-HT2C mutant mice. In contrast, mutant mice showed an increased sensitivity to the stereotypy induced by high doses of d-fenfluramine (10, 30 mg/kg) compared to that of wild-type littermates. These data demonstrate a role for the 5-HT2C receptor in mediating d-fenfluramine-induced satiety.


How medical students learn ethics: an online log of their learning experiences.
Medical students experience ethics learning in a wide variety of formats, delivered not just through the taught curriculum. An audit of ethics learning was carried out at a medical school through a secure website over one academic year to determine the quantity and range of medical ethics learning in the undergraduate curriculum and compare this with topics for teaching described by the Institute of Medical Ethics (IME) (2010) and the General Medical Council's (GMC) Tomorrow's Doctors (2009). The online audit captured the participants' reflections on their learning experiences and the impact on their future practice. Results illustrate the opportunistic nature of ethics learning, especially in the clinical years, and highlight the reality of the hidden curriculum for medical students. Overall, the ethics learning was a helpful and positive experience for the participants and fulfils the GMC and IME curriculum requirements.


The chromatin remodeler Mi-2beta is required for CD4 expression and T cell development.
Changes in chromatin structure underlie the activation or silencing of genes during development. The chromatin remodeler Mi-2beta is highly expressed in thymocytes and is presumed to be a transcriptional repressor because of its presence in the nucleosome remodeling deacetylase (NuRD) complex. Using conditional inactivation, we show that Mi-2beta is required at several steps during T cell development: for differentiation of beta selected immature thymocytes, for developmental expression of CD4, and for cell divisions in mature T cells. We further show that Mi-2beta plays a direct role in promoting CD4 gene expression. Mi-2beta associates with the CD4 enhancer as well as the E box binding protein HEB and the histone acetyltransferase (HAT) p300, enabling their recruitment to the CD4 enhancer and causing histone H3-hyperacetylation to this regulatory region. These findings provide important insights into the regulation of CD4 expression during T cell development and define a role for Mi-2beta in gene activation.


How Should We Follow-Up Asymptomatic Metal-on-Metal Hip Resurfacing Patients? A Prospective Longitudinal Cohort Study.
Current surveillance for metal-on-metal hip resurfacing (MoMHR) patients is not evidence based. This study established changes that occurred in 152 asymptomatic MoMHRs using repeat ultrasound and patient-reported outcomes. Factors associated with (1) ultrasound progression and (2) developing new pseudotumors were analyzed. Patients underwent repeat assessments 4.3 years later. Ultrasound progression was observed in 19% (n = 29), with 10% (n = 15) developing new pseudotumors. Key predictors of ultrasound progression included high blood cobalt (P = .00013) and chromium (P = .00065), and high initial ultrasound grade (P = .003) and volume (P = .036). No asymptomatic MoMHRs with initially normal metal ions (<2 μg/L) and normal ultrasounds (33% of cohort) developed new pseudotumors. This patient subgroup does not require repeat follow-up within 5 years.


Human serum albumin redox state is associated with decreased renal function in a community-dwelling population.
The redox state of human serum albumin (HSA) has attracted interest as a possible biomarker for oxidative stress (OS) in humans. Although previous studies on this topic have taken only clinical settings into consideration, evidence of its efficacy in nonclinical settings remains to be established. The present study aimed to examine and validate the relationship between HSA redox state and renal function in a rural Japanese population. We analyzed two independent data sets from health checkup programs conducted in 2013 and 2016: one for discovery ( n = 267) and the other for replication ( n = 367). The fraction of human mercaptalbumin (HMA) to total HSA [f(HMA)] was determined using our revised method of high-performance liquid chromatography with post-column bromocresol green. The estimated glomerular filtration rate (eGFR) was calculated based on each individual's serum creatinine value, sex, and age. Adjustment for potential confounders revealed positive associations of fraction of human mercaptalbumin [f(HMA)] with eGFR in the discovery and replication analyses ( P < 0.001 and P = 0.03, respectively). Multivariate logistic regression analyses demonstrated significant inverse associations between renal dysfunction (defined as eGFR < 60 ml·min-1·1.73 m-2) and f(HMA) by a factor of 0.50 and 0.65 (confidence intervals of 0.26-0.91 and 0.37-1.00), respectively, with a unit of 10% f(HMA). Our results indicate that HSA redox state is consistently associated with renal dysfunction in both clinical and nonclinical settings.


A novel low-molecular-weight compound enhances ectopic bone formation and fracture repair.
Use of recombinant human bone morphogenetic protein-2 (rhBMP-2) is expensive and may cause local side effects. A small synthetic molecule, SVAK-12, has recently been shown in vitro to potentiate rhBMP-2-induced transdifferentiation of myoblasts into the osteoblastic phenotype. The aims of this study were to test the ability of SVAK-12 to enhance bone formation in a rodent ectopic model and to test whether a single percutaneous injection of SVAK-12 can accelerate callus formation in a rodent femoral fracture model. Collagen disks with rhBMP-2 alone or with rhBMP-2 and SVAK-12 were implanted in a standard athymic rat chest ectopic model, and radiographic analysis was performed at four weeks. In a second set of rats (Sprague-Dawley), SVAK-12 was percutaneously injected into the site of a closed femoral fracture. The fractures were analyzed radiographically and biomechanically (with torsional testing) five weeks after surgery. In the ectopic model, there was dose-dependent enhancement of rhBMP-2 activity with use of SVAK-12 at doses of 100 to 500 μg. In the fracture model, the SVAK-12-treated group had significantly higher radiographic healing scores than the untreated group (p = 0.028). Biomechanical testing revealed that the fractured femora in the 200 to 250-μg SVAK-12 group were 43% stronger (p = 0.008) and 93% stiffer (p = 0.014) than those in the control group. In summary, at five weeks the femoral fracture group injected with SVAK-12 showed significantly improved radiographic and biomechanical evidence of healing compared with the controls. A single local dose of a low-molecular-weight compound, SVAK-12, enhanced bone-healing in the presence of low-dose exogenous rhBMP-2 (in the ectopic model) and endogenous rhBMPs (in the femoral fracture model). This study demonstrates that rhBMP-2 responsiveness can be enhanced by a novel small molecule, SVAK-12. Local application of anabolic small molecules has the potential for potentiating and accelerating fracture-healing. Use of this small molecule to lower required doses of rhBMPs might both decrease their cost and improve their safety profile.


How Neighborhoods Influence Health: Lessons to be learned from the application of political ecology.
This paper articulates how political ecology can be a useful tool for asking fundamental questions and applying relevant methods to investigate structures that impact relationship between neighborhood and health. Through a narrative analysis, we identify how political ecology can develop our future agendas for neighborhood-health research as it relates to social, political, environmental, and economic structures. Political ecology makes clear the connection between political economy and neighborhood by highlighting the historical and structural processes that produce and maintain social inequality, which affect health and well-being. These concepts encourage researchers to examine how people construct neighborhood and health in different ways that, in turn, can influence different health outcomes and, thus, efforts to address solutions.


Sitting tai chi improves the balance control and muscle strength of community-dwelling persons with spinal cord injuries: a pilot study.
Objective. To investigate the effects of sitting Tai Chi on muscle strength, balance control, and quality of life (QOL) among survivors with spinal cord injuries (SCI). Methods. Eleven SCI survivors participated in the sitting Tai Chi training (90 minutes/session, 2 times/week for 12 weeks) and eight SCI survivors acted as controls. Dynamic sitting balance was evaluated using limits of stability test and a sequential weight shifting test in sitting. Handgrip strength was also tested using a hand-held dynamometer. QOL was measured using the World Health Organization's Quality of Life Scale. Results. Tai Chi practitioners achieved significant improvements in their reaction time (P = 0.042); maximum excursion (P = 0.016); and directional control (P = 0.025) in the limits of stability test after training. In the sequential weight shifting test, they significantly improved their total time to sequentially hit the 12 targets (P = 0.035). Significant improvement in handgrip strength was also found among the Tai Chi practitioners (P = 0.049). However, no significant within and between-group differences were found in the QOL outcomes (P > 0.05). Conclusions. Twelve weeks of sitting Tai Chi training could improve the dynamic sitting balance and handgrip strength, but not QOL, of the SCI survivors.


A new painkiller nanomedicine to bypass the blood-brain barrier and the use of morphine.
The clinical use of endogenous neuropeptides has historically been limited due to pharmacokinetic issues, including plasma stability and blood-brain barrier permeability. In this study, we show that the rapidly metabolized Leu-enkephalin (LENK) neuropeptide may become pharmacologically efficient owing to a simple conjugation with the lipid squalene (SQ). The corresponding LENK-SQ bioconjugates were synthesized using different chemical linkers in order to modulate the LENK release after their formulation into nanoparticles. This new SQ-based nanoformulation prevented rapid plasma degradation of LENK and conferred on the released neuropeptide a notable antihyperalgesic effect that lasted longer than after treatment with morphine in a rat model of inflammation (Hargreaves test). The biodistribution study as well as the use of brain-permeant and -impermeant opioid receptor antagonists indicated that LENK-SQ NPs act through peripherally located opioid receptors. This study represents a novel nanomedicine approach, allowing the specific delivery of LENK neuropeptide into inflamed tissues for pain control.


The evolutionary genetics and emergence of avian influenza viruses in wild birds.
We surveyed the genetic diversity among avian influenza virus (AIV) in wild birds, comprising 167 complete viral genomes from 14 bird species sampled in four locations across the United States. These isolates represented 29 type A influenza virus hemagglutinin (HA) and neuraminidase (NA) subtype combinations, with up to 26% of isolates showing evidence of mixed subtype infection. Through a phylogenetic analysis of the largest data set of AIV genomes compiled to date, we were able to document a remarkably high rate of genome reassortment, with no clear pattern of gene segment association and occasional inter-hemisphere gene segment migration and reassortment. From this, we propose that AIV in wild birds forms transient "genome constellations," continually reshuffled by reassortment, in contrast to the spread of a limited number of stable genome constellations that characterizes the evolution of mammalian-adapted influenza A viruses.


Active control of the strong coupling regime between porphyrin excitons and surface plasmon polaritons.
We experimentally demonstrate the active control of the coupling strength between porphyrin dyes and surface plasmon polaritons supported by a thin gold layer. This control is externally exerted by a gas flow and is reversible. The hybridized exciton-polariton branches resulting from the exciton-plasmon coupling display a splitting proportional to the coupling strength of the light-matter interaction. The coupled system changes from the weak (no splitting) to the strong coupling regime (splitting of 130 meV) by controlling the effective oscillator strength in the dye layer, via exposure to nitrogen dioxide. The modification of the coupling strength of the system allows tailoring of the dispersion of the hybridized modes as well as of their group velocity.


Caveolins and cavins in the trafficking, maturation, and degradation of caveolae: implications for cell physiology.
Caveolins (Cavs) are ~20 kDa scaffolding proteins that assemble as oligomeric complexes in lipid raft domains to form caveolae, flask-shaped plasma membrane (PM) invaginations. Caveolae ("little caves") require lipid-lipid, protein-lipid, and protein-protein interactions that can modulate the localization, conformational stability, ligand affinity, effector specificity, and other functions of proteins that are partners of Cavs. Cavs are assembled into small oligomers in the endoplasmic reticulum (ER), transported to the Golgi for assembly with cholesterol and other oligomers, and then exported to the PM as an intact coat complex. At the PM, cavins, ~50 kDa adapter proteins, oligomerize into an outer coat complex that remodels the membrane into caveolae. The structure of caveolae protects their contents (i.e., lipids and proteins) from degradation. Cellular changes, including signal transduction effects, can destabilize caveolae and produce cavin dissociation, restructuring of Cav oligomers, ubiquitination, internalization, and degradation. In this review, we provide a perspective of the life cycle (biogenesis, degradation), composition, and physiologic roles of Cavs and caveolae and identify unanswered questions regarding the roles of Cavs and cavins in caveolae and in regulating cell physiology.1.


Association of Serum Tocilizumab Trough Concentrations with Clinical Disease Activity Index Scores in Adult Patients with Rheumatoid Arthritis.
To determine whether serum trough concentrations of tocilizumab (TCZ) administered as a fixed-dose subcutaneous (SC) injection for the treatment of rheumatoid arthritis (RA) are associated with disease activity responses. We analyzed datasets from the Israeli branch of the multinational TOZURA study, which evaluated a weekly subcutaneous TCZ treatment regimen in a real-life clinical setting. Generalized estimating equations (GEE) were used to evaluate associations between the TCZ levels and the study outcomes. Linear models and GEE were used to evaluate associations between patient characteristics and TCZ levels. A significant association between the TCZ concentrations and the change in the Clinical Disease Activity Index (CDAI) score was observed. In a multivariate binary GEE model, every increase of 10 µg/ml in the concentration of TCZ was associated with being in a state of CDAI remission or low disease activity (OR 1.41) versus a moderate/high disease activity state. An OR of 1.52 was associated with being in a state of Health Assessment Questionnaire-Disability Index remission. In univariate linear models, there was an inverse association between body mass index (BMI) and improvement in the CDAI score, and the BMI score was associated with lower TCZ concentrations. Patients who weighed > 100 kg had lower TCZ concentrations. In the first 24 weeks of treatment with SC TCZ injections, TCZ concentrations were associated with clinical improvement, while body weight and BMI were inversely associated with TCZ concentrations. Personalizing the dose of SC TCZ to body weight may improve outcomes of clinical disease activity in patients with RA.


PinX1, a telomere repeat-binding factor 1 (TRF1)-interacting protein, maintains telomere integrity by modulating TRF1 homeostasis, the process in which human telomerase reverse Transcriptase (hTERT) plays dual roles.
TRF1, a telomere-binding protein, is important for telomere protection and homeostasis. PinX1 interacts with TRF1, but the physiological consequences of their interaction in telomere protection are not yet understood. Here we investigated PinX1 function on TRF1 stability in HeLa cells. PinX1 overexpression stabilized TRF1, but PinX1 depletion by siRNA led to TRF1 degradation, TRF1 ubiquitination, and less TRF1 telomere association. The depletion also induced DNA damage responses at telomeres and chromosome instability. These telomere dysfunctional phenotypes were in fact due to TRF1 deficiency. We also report that hTERT, a catalytic component of telomerase, plays dual roles in the TRF1 steady state pathway. PinX1-mediated TRF1 stability was not observed in hTERT-negative immortal cells, but was pronounced when hTERT was ectopically expressed in the cells, suggesting that hTERT may be needed in the PinX1-mediated TRF1 stability pathway. Interestingly, the knockdown of both PinX1 and hTERT in HeLa cells stabilized TRF1, suppressed DNA damage response activation, and restored chromosome stability. In summary, our findings suggested that PinX1 may maintain telomere integrity by regulating TRF1 stability and that hTERT may act as both a positive and a negative regulator of TRF1 homeostasis in a PinX1-dependent manner.


Implementation of phase-shift patterns using a holographic projection system with phase-only diffractive optical elements.
We proposed a method to implement spatial phase-shift patterns with subdiffraction limited features through a holographic projection system. The input device of the system displayed phase-only diffractive optical elements that were calculated using the iterative Fourier-transform algorithm with the dummy-area method. By carefully designing the target patterns to the algorithm, the diffractive optical elements generated the Fourier-transformed images containing the phase-shift patterns in which the widths of dark lines were smaller than the diffraction limit. With these demonstrations, we have successfully shown that the near-field phase-shift lithographic technique can be realized through an inexpensive maskless lithographic system and can still achieve subdiffraction limited images.