Pelvic-floor muscle rehabilitation in erectile dysfunction and premature ejaculation.
In men, involuntary or voluntary ischiocavernosus muscle contractions after erection lead to intracavernous blood pressures far higher than the systolic pressure, which builds and maintains penile rigidity. Thus, erectile dysfunction may be partly due to ischiocavernosus muscle atrophy and may be treated by rehabilitation interventions. The purpose of this study was to determine whether pelvic-floor muscle strengthening interventions could be associated with increases in intracavernous pressure that would increase penile rigidity. An observational study was conducted. One hundred twenty-two men with isolated erectile dysfunction and 108 men with isolated premature ejaculation participated (no neuromuscular diseases or previous perineal rehabilitation). Thirty-minute sessions of voluntary contractions coupled with electrical stimulation were designed to increase ischiocavernosus muscle strength (monitored through intracavernous pressure increase). A linear mixed-effects model per group analyzed separately, then jointly, the maximum change in pressure (ΔP) and the maximum baseline (ie, respectively, the average contraction-generated difference in intracavernous pressure and the intracavernous pressure plateau at full erection, both measured during the highest moving average of the best 2 minutes of each session). Over 20 sessions, the maximum ΔP increased in erectile dysfunction as well as in premature ejaculation (87% and 88%, respectively, in men with positive trends). The maximum baseline also increased (99% and 72%, respectively, in men with positive trends). The joint modeling indicated that the mean expected progressions of the intracavernous pressure after 5 sessions in erectile dysfunction and premature ejaculation were 62.85 and 64.15 cm H2O, respectively. Indirect measurements were obtained of intracavernous pressure and ischiocavernosus muscle force. Pelvic-floor muscle rehabilitation was found to be beneficial in erectile dysfunction. However, its effects on symptoms of premature ejaculation, despite intracavernous pressure gains, were much more difficult to assess. The definitive proof of its benefits requires rather difficult-to-design clinical trials.


Arcobacter cryaerophilus Isolated From New Zealand Mussels Harbor a Putative Virulence Plasmid.
A wide range of Arcobacter species have been described from shellfish in various countries but their presence has not been investigated in Australasia, in which shellfish are a popular delicacy. Since several arcobacters are considered to be emerging pathogens, we undertook a small study to evaluate their presence in several different shellfish, including greenshell mussels, oysters, and abalone (paua) in New Zealand. Arcobacter cryaerophilus, a species associated with human gastroenteritis, was the only species isolated, from greenshell mussels. Whole-genome sequencing revealed a range of genomic traits in these strains that were known or associated virulence factors. Furthermore, we describe the first putative virulence plasmid in Arcobacter, containing lytic, immunoavoidance, adhesion, antibiotic resistance, and gene transfer traits, among others. Complete genome sequence determination using a combination of long- and short-read genome sequencing strategies, was needed to identify the plasmid, clearly identifying its benefits. The potential for plasmids to disseminate virulence traits among Arcobacter and other species warrants further consideration by researchers interested in the risks to public health from these organisms.


Genetics of nurture: A test of the hypothesis that parents' genetics predict their observed caregiving.
Twin studies have documented that parenting behavior is partly heritable, but it is unclear how parents' genetics shape their caregiving. Using tools of molecular genetics, the present study investigated this process by testing hypotheses about associations between a genome-wide polygenic score for educational attainment and parental caregiving in 702 members of the Dunedin Study, a population-representative birth cohort. Data have been prospectively collected from when Study members were born through to midlife, and include assessments of the caregiving they provided once they became parents. Results showed that parents' polygenic scores predicted warm, sensitive, and stimulating caregiving, both in personal interactions with their young children (as captured on video) and through the home environments they created for their families (as observed by home visitors). The magnitude of this effect was small. Polygenic-score associations were independent of well-established predictors of parenting, such as parents' own childhood experiences of parenting and the age at which they became parents. Polygenic-score associations were mediated by parents' early-emerging cognitive abilities and self-control skills. Findings have implications for theory and research about genetic influences on caregiving and child development. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Retrotransposon long interspersed nucleotide element-1 (LINE-1) is activated during salamander limb regeneration.
Salamanders possess an extraordinary capacity for tissue and organ regeneration when compared to mammals. In our effort to characterize the unique transcriptional fingerprint emerging during the early phase of salamander limb regeneration, we identified transcriptional activation of some germline-specific genes within the Mexican axolotl (Ambystoma mexicanum) that is indicative of cellular reprogramming of differentiated cells into a germline-like state. In this work, we focus on one of these genes, the long interspersed nucleotide element-1 (LINE-1) retrotransposon, which is usually active in germ cells and silent in most of the somatic tissues in other organisms. LINE-1 was found to be dramatically upregulated during regeneration. In addition, higher genomic LINE-1 content was also detected in the limb regenerate when compared to that before amputation indicating that LINE-1 retrotransposition is indeed active during regeneration. Active LINE-1 retrotransposition has been suggested to have a potentially deleterious impact on genomic integrity. Silencing of activated LINE-1 by small RNAs has been reported to be part of the machinery aiming to maintain genomic integrity. Indeed, we were able to identify putative LINE-1-related piRNAs in the limb blastema. Transposable element-related piRNAs have been identified frequently in the germline in other organisms. Thus, we present here a scenario in which a unique germline-like state is established during axolotl limb regeneration, and the re-activation of LINE-1 may serve as a marker for cellular dedifferentiation in the early-stage of limb regeneration.


Muscle coenzyme Q10 level in statin-related myopathy.
Statin drugs (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) reduce the level of cholesterol by inhibiting the synthesis of mevalonate, an intermediary in the cholesterol biosynthetic pathway. Use of statin drugs has been associated with a variety of skeletal muscle-related complaints. Coenzyme Q10 (CoQ10), a component of the mitochondrial respiratory chain, is also synthesized from mevalonate, and decreased muscle CoQ10 concentration may have a role in the pathogenesis of statin drug-related myopathy. To measure the CoQ10 concentration and respiratory chain enzyme activities in muscle biopsy specimens from 18 patients with statin drug-related myopathy and to look for evidence of apoptosis using the TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling) assay. An open-labeled study of CoQ10 concentration in muscle from patients with increased serum creatine kinase concentrations while receiving standard statin drug therapy. Neuromuscular centers at 2 academic tertiary care hospitals. Muscle structure was essentially normal in 14 patients and showed evidence of mitochondrial dysfunction and nonspecific myopathic changes in 2 patients each. Muscle CoQ10 concentration was not statistically different between patients and control subjects, but it was more than 2 SDs below the normal mean in 3 patients and more than 1 SD below normal in 7 patients. There was no TUNEL positivity in any patients. These data suggest that statin drug-related myopathy is associated with a mild decrease in muscle CoQ10 concentration, which does not cause histochemical or biochemical evidence of mitochondrial myopathy or morphologic evidence of apoptosis in most patients.


Attitudes and norms affecting scientists' data reuse.
The value of sharing scientific research data is widely appreciated, but factors that hinder or prompt the reuse of data remain poorly understood. Using the Theory of Reasoned Action, we test the relationship between the beliefs and attitudes of scientists towards data reuse, and their self-reported data reuse behaviour. To do so, we used existing responses to selected questions from a worldwide survey of scientists developed and administered by the DataONE Usability and Assessment Working Group (thus practicing data reuse ourselves). Results show that the perceived efficacy and efficiency of data reuse are strong predictors of reuse behaviour, and that the perceived importance of data reuse corresponds to greater reuse. Expressed lack of trust in existing data and perceived norms against data reuse were not found to be major impediments for reuse contrary to our expectations. We found that reported use of models and remotely-sensed data was associated with greater reuse. The results suggest that data reuse would be encouraged and normalized by demonstration of its value. We offer some theoretical and practical suggestions that could help to legitimize investment and policies in favor of data sharing.


Indications for surgical management of osteochondritis dissecans of the knee in the pediatric population: a systematic review.
Several case series have been published exploring the surgical management of osteochondritis dissecans (OCD) of the knee in pediatric patients. This systemic review was performed to identify the surgical indications for this condition. A search of the Embase and Ovid Medline databases was performed to identify clinical studies reporting outcomes of surgical management of OCD in the knee in this patient population. A quality assessment of the included articles was conducted independently by two reviewers using a quality assessment tool developed by Yang et al. A total of 25 articles met the eligibility criteria and were reviewed; 40% of studies did not clearly describe their surgical indications. The remainder of the studies had a failure of nonoperative management with or without the concomitant use of imaging as their indication for surgery, or used lesion stability itself as the indication for surgery. This review outlines several surgical indications presented in the literature for the treatment of OCD lesions of the knee in the pediatric population. The most common indication for surgery was a failure of a trial of nonoperative treatment with or without the concomitant use of serial imaging. Although the quality of the case series was high, inconsistencies in reporting radiographic and arthroscopic classification of the OCD lesion were common.


Optimizing Decision-Making Processes in Times of COVID-19: Using Reflexivity to Counteract Information-Processing Failures.
The effectiveness of policymakers' decision-making in times of crisis depends largely on their ability to integrate and make sense of information. The COVID-19 crisis confronts governments with the difficult task of making decisions in the interest of public health and safety. Essentially, policymakers have to react to a threat, of which the extent is unknown, and they are making decisions under time constraints in the midst of immense uncertainty. The stakes are high, the issues involved are complex and require the careful balancing of several interests, including (mental) health, the economy, and human rights. These circumstances render policymakers' decision-making processes vulnerable to errors and biases in the processing of information, thereby increasing the chances of faulty decision-making processes with poor outcomes. Prior research has identified three main information-processing failures that can distort group decision-making processes and can lead to negative outcomes: (1) failure to search for and share information, (2) failure to elaborate on and analyze information that is not in line with earlier information and (3) failure to revise and update conclusions and policies in the light of new information. To date, it has not yet been explored how errors and biases underlying these information-processing failures impact decision-making processes in times of crisis. In this narrative review, we outline how groupthink, a narrow focus on the problem of containing the virus, and escalation of commitment may pose real risks to decision-making processes in handling the COVID-19 crisis and may result in widespread societal damages. Hence, it is vital that policymakers take steps to maximize the quality of the decision-making process and increase the chances of positive outcomes as the crisis goes forward. We propose group reflexivity-a deliberate process of discussing team goals, processes, or outcomes-as an antidote to these biases and errors in decision-making. Specifically, we recommend several evidence-based reflexivity tools that could easily be implemented to counter these information-processing errors and improve decision-making processes in uncertain times.


Tissue-specific patterns of a maize Myb transcription factor are epigenetically regulated.
The maize p1 gene encodes a Myb-homologous regulator of red pigment biosynthesis. To investigate the tissue-specific regulation of the p1 gene, maize plants were transformed with constructs combining promoter and cDNA sequences of two alleles which differ in pigmentation patterns: P1-wr (white pericarp/red cob) and P1-rr (red pericarp/red cob). Surprisingly, all promoter/cDNA combinations produced transgenic plants with red pericarp and red cob (RR pattern), indicating that the P1-wr promoter and encoded protein can function in pericarp. Some of the RR patterned transgenic plants produced progeny plants with white pericarp and red cob (WR pattern), and this switch in tissue-specificity correlated with increased transgene methylation. A similar inverse correlation between pericarp pigmentation and DNA methylation was observed for certain natural p1 alleles, which have a gene structure characteristic of standard P1-wr alleles, but which confer red pericarp pigmentation and are consistently less methylated than standard P1-wr alleles. Although we cannot rule out the possible existence of tissue-specific regulatory elements within the p1 non-coding sequences or flanking regions, the data from transgenic and natural alleles suggest that the tissue-specific pigmentation pattern characteristic of the P1-wr phenotype is epigenetically controlled.


Morphology-dependent resonances of nearly spherical particles in random orientation.
We use precise T-matrix calculations for prolate and oblate spheroids, Chebyshev particles, and spheres cut by a plane to study the evolution of Lorenz-Mie morphology-dependent resonances (MDRs) with increasing asphericity of nearly spherical particles in random orientation. We show that, in the case of spheroids and Chebyshev particles, the deformation of a sphere by as little as one hundredth of a wavelength essentially annihilates supernarrow MDRs, whereas significantly larger asphericities are needed to suppress broader resonance features. The MDR position and profile are also affected when the deviation of the particle shape is increased from that of a perfect sphere. In the case of a sphere cut by a plane, the supernarrow MDRs are much more resistant to an increase in asphericity and do not change their position and profile. These findings are consistent with the widely accepted physical interpretation of the Lorenz-Mie MDRs.


Under-contouring of rods: a potential risk factor for proximal junctional kyphosis after posterior correction of Scheuermann kyphosis.
Scheuermann kyphosis (SK) could require surgical treatment in certain situations. A posterior reduction is the most widespread treatment so far, although the development of proximal junctional kyphosis (PJK) is one of the possible complications of this procedure. The contour of the proximal part of the rod could influence the occurrence of PJK in SK patients. The objective of this study was to analyze the impact of the proximal rod contour on the occurrence of a PJK complication in SK patients. This retrospective monocentric study was performed in the Nanjing Spine Surgery Department. All eligible patients had undergone posterior correction surgery with pedicle screws only between 2002 and 2017 and had at least 24 months of follow-up. The presence of PJK was quantified on radiographs using the proximal junctional angle (PJA > 10° at the last follow-up). The authors propose a new radiological parameter to measure the angulation of the proximal part of the instrumentation: the proximal contouring rod angle (PCRA) is the angle between the upper endplate of the upper instrumented vertebra (UIV) and the lower endplate of the second vertebra caudal to the UIV. The patients were analyzed according to the presence or absence of PJK. A t-test, receiver operating characteristic (ROC) curve analysis, and logistic regression analysis were performed for statistical analysis. Sixty-two patients treated for SK were included in this study. The mean age was 18.6 ± 8.5 years, and the mean follow-up was 42.5 ± 16.4 months. The mean correction rate of global kyphosis was 46.4% ± 13.7%. At the last follow-up, 17 patients (27.4%) presented with PJK. No significant difference was found between the PJK and non-PJK groups in terms of age and other preoperative variables. A significant difference in the postoperative PCRA was found between the PJK and non-PJK groups (8.2° ± 4.9° vs 15.7° ± 6.6°, respectively; p = 0.001). A postoperative PCRA less than 10.1° predicted a significantly higher risk for PJK (p = 0.002, OR 2.431, 95% CI 1.781-4.133). Under-contouring of the proximal part of the rods (lower than 10°) is a risk factor for PJK after posterior correction of SK.


Pacemakers and magnetic resonance imaging: Current status and survey in Switzerland.
Electromagnetic fields arising from magnetic resonance imaging (MRI) can cause various clinically relevant functional disturbances in patients with cardiac pacemakers. Consequently, an implanted pacemaker is generally considered a contraindication for an MRI scan. With approximately 60 million MRI scans performed worldwide per year, MRI may be indicated for an estimated majority of pacemaker patients during the lifetime of their pacemakers. The availability of MR conditional pacemakers with CE labelling is of particular advantage since they allow the safe use of pacemakers in MRI. In this article the current state of knowledge on pacemakers and MR imaging is discussed. We present the results of a survey conducted among Swiss radiologists to assess current practice in patients with pacemakers.


In vitro protective effects of colon-available extract of Camellia sinensis (tea) against hydrogen peroxide and beta-amyloid (Aβ((1-42))) induced cytotoxicity in differentiated PC12 cells.
There is mounting evidence that the deposition and aggregation of β-amyloid peptides (Aβ) in the brain play a significant role in the development and pathogenesis of Alzheimer's disease. There is further evidence that free radical species such as hydrogen peroxide (H(2)O(2)) mediate Aβ induced toxicity. Previous studies have demonstrated that green tea polyphenols possess neuroprotective properties through their ability to ameliorate oxidative stress induced by free radical species. Green tea polyphenols have also been shown to enhance cognition in various animal models of induced cognitive impairment. Upon ingestion, green tea polyphenols are metabolised and undergo bio-transformation which affects their bioavailability and therefore efficacy. In this study, a green tea extract was subjected to a simulated gastrointestinal digestion and a 'colon-available' extract (CAGTE) prepared and assessed for its potential protective effects against H(2)O(2) and Aβ((1-42)) induced cytotoxicity using differentiated PC12 cells (dPC12) as a model for neuronal cells. CAGTE represents green tea phytochemicals potentially available after upper gastrointestinal digestion. CAGTE which was depleted in flavan-3-ols, as shown by LC-MS analysis, protected dPC12 cells at concentration ranges of 0.3-10 μg/ml and 0.03-0.125 μg/ml for H(2)O(2) and Aβ((1-42)), induced cytotoxicity, respectively. At high concentrations, CAGTE exhibited direct anti-proliferative effects, in line with the reputed anti-cancer properties of green tea polyphenols. These results demonstrate that potentially bioavailable green tea metabolites are able to ameliorate both H(2)O(2) and Aβ((1-42)) induced cytotoxicity.


A Bacterial Artificial Chromosome Reporter System for Expression of the Human FOXP3 Gene in Mouse Regulatory T-Cells.
The transcription factor FOXP3 plays key roles in the development and function of regulatory T cells (Treg) capable of preventing and correcting immunopathology. There has been much interest in exploiting Treg as adoptive cell therapy in man, but issues of lack of nominal antigen-specificity and stability of FoxP3 expression in the face of pro-inflammatory cytokines have been a concern. In order to enable fundamental studies of human FOXP3 (hFOXP3) gene regulation and to provide preclinical tools to guide the selection of drugs that might modulate hFOXP3 expression for therapeutic purposes, we generated hFOXP3/AmCyan bacterial artificial chromosome (BAC) transgenic mice and transfectants, wherein hFOXP3 expression was read out as AmCyan expression. Using the transgenic mice, one can now investigate hFOXP3 gene expression under defined experimental conditions used for mouse Foxp3 (mFoxp3) studies. Here, we demonstrate that hFOXP3 gene expression in BAC transgenic mice is solely restricted to CD4+ T-cells, as for mFoxp3 gene expression, showing that hFOXP3 expression in Treg cells depends on fundamentally similar processes to mFoxp3 expression in these cells. Similarly, hFOXP3 expression could be observed in mouse T-cells through TCR stimulation in the presence of TGF-β. These data suggest that, at least in part, cell type-specific human and mouse foxp3 gene expression is regulated by common regulatory regions which for the human, are located within the 110-kb human FOXP3 BAC DNA. To investigate hFOXP3 gene expression further and to screen potential therapeutics in modulating hFOXP3 gene expression in vitro, we also generated hFOXP3/AmCyan expression reporter cell lines. Using the reporter cells and transcription factor inhibitors, we showed that, just as for mFoxp3 expression, inhibitors of NF-κB, AP1, STAT5, Smad3, and NFAT also block hFOXP3 expression. hFOXP3 induction in the reporter cells was also TGF-β dependent, and substantially enhanced by an mTOR inhibitor, Torin1. In both the reporter transgenic mice and cell lines, histone H4 molecules in the hFOXP3 promoter and enhancers located in human CNS1 and CNS2 regions were highly acetylated in natural Treg and TCR/TGF-β-induced Treg, indicating hFOXP3 gene expression is regulated by mechanisms similar to those previously identified for the mFoxp3 gene.


Myalgic encephalomyelitis: International Consensus Criteria.
The label 'chronic fatigue syndrome' (CFS) has persisted for many years because of the lack of knowledge of the aetiological agents and the disease process. In view of more recent research and clinical experience that strongly point to widespread inflammation and multisystemic neuropathology, it is more appropriate and correct to use the term 'myalgic encephalomyelitis' (ME) because it indicates an underlying pathophysiology. It is also consistent with the neurological classification of ME in the World Health Organization's International Classification of Diseases (ICD G93.3). Consequently, an International Consensus Panel consisting of clinicians, researchers, teaching faculty and an independent patient advocate was formed with the purpose of developing criteria based on current knowledge. Thirteen countries and a wide range of specialties were represented. Collectively, members have approximately 400 years of both clinical and teaching experience, authored hundreds of peer-reviewed publications, diagnosed or treated approximately 50 000 patients with ME, and several members coauthored previous criteria. The expertise and experience of the panel members as well as PubMed and other medical sources were utilized in a progression of suggestions/drafts/reviews/revisions. The authors, free of any sponsoring organization, achieved 100% consensus through a Delphi-type process. The scope of this paper is limited to criteria of ME and their application. Accordingly, the criteria reflect the complex symptomatology. Operational notes enhance clarity and specificity by providing guidance in the expression and interpretation of symptoms. Clinical and research application guidelines promote optimal recognition of ME by primary physicians and other healthcare providers, improve the consistency of diagnoses in adult and paediatric patients internationally and facilitate clearer identification of patients for research studies.


Injection risk norms and practices among migrant Puerto Rican people who inject drugs in New York City: The limits of acculturation theory.
Among people who inject drugs (PWID) in New York City (NYC), racial minorities are disproportionately infected with HIV and hepatitis C (HCV). Prior research has shown that PWID who started injecting drugs in Puerto Rico (P.R.) tend to maintain the risky injection behaviors learned there. This study identifies the P.R.-native norms supporting the continued injection risk behavior of migrant Puerto Rican PWID in NYC to inform a culturally appropriate risk-reduction intervention. 40 migrant Puerto Rican PWID were recruited in NYC for a longitudinal qualitative study. The sample was stratified to include 20 migrants with <3 years in NYC and 20 migrants with >3-6 years in NYC. Time-location sampling was used to curb possible network bias in recruitment. Over 12 months, migrants completed semi-structured interviews at baseline, monthly follow-ups, and study exit. Analyses were guided by grounded theory. Most participants (90%) reported having had chronic HCV, and 22.5% reported being HIV-positive. Syringe- and cooker-/cotton-sharing were widespread in both P.R. and NYC. The ubiquitous practice of cleaning used syringes by "water-rinsing and air-blowing" was guided by a normative belief, learned in P.R., that "water and air kill HIV." Sterile syringe use was not a priority. HCV was not a concern. P.R.-native abstinence-only narratives discouraged opioid agonist treatment (OAT) enrollment among recent migrants (≤3 years). Experiences with drug dealers, prison-power groups, and injection doctors ("Gancheros") in P.R. influenced migrants' injection risk behavior in NYC. Those who were Gancheros in P.R. continued working as Gancheros in NYC. Injection risks make migrant Puerto Rican PWID in NYC vulnerable to HIV/HCV. Harm reduction programs should pay closer attention to the rationales behind these injection risks. A risk-reduction intervention that incorporates the Ganchero figure may be a credible way to help migrants reduce injection risk and accept OAT and syringe exchange programs (SEP).


Cellular mechanisms in inflammatory myopathies.
Cell-mediated immune mechanisms play a prominent role in inclusion body myositis (IBM) and polymyositis (PM). In both IBM and PM, CD8+ cytotoxic T cells expressing the alpha/beta receptor surround and focally invade non-necrotic muscle fibres. This lesion can be considered the hallmark of cell-mediated myocytotoxicity. Essentially the same type of lesion is observed in a variant form of PM, in which CD4-CD8- T cells bearing the gamma/delta receptor surround and invade non-necrotic muscle fibres. In both IBM and PM, all of the invaded and some of the non-invaded muscle fibres strongly express HLA class I molecules. This is consistent with the hypothesis that the CD8+ autoinvasive cytotoxic T cells recognize antigenic peptide(s) bound to HLA class I molecules on the muscle fibre surface. According to the rules of antigen processing, these peptides derive from proteins synthesized in the muscle fibre. Theoretically, the proteins could be viral components or self proteins that resemble viral components. Most attempts to demonstrate viral antigens or genome in muscle fibres have failed. On the other hand, the majority of HLA class I molecules expressed on the surface of any cell are loaded with endogenous self peptides. It seems plausible that muscle-specific autoantigen(s) could be recognized by autoaggressive T cells in the inflammatory myopathies, but the precise reasons for the recognition event remains elusive. Recently, it has become possible to study the interactions of muscle cells and cytotoxic effector cells in vitro. Myoblasts and myotubes can be induced to express a variety of immunologically relevant histocompatibility and cell adhesion molecules. Myotubes are highly susceptible to lysis by allogeneic CD8+ cytotoxic T cells sensitized against HLA class I alloantigens. Interestingly, cultured myotubes are also susceptible to lysis by antigen-nonspecific natural killer cells. Further, myoblasts stimulated by IFN gamma express HLA class II and acquire the full potential to process and present complex protein antigens to CD4+ T cells. This may indicate that myoblasts can actively participate in local immune reactions by presenting (auto) antigens to helper/inducer T cells. In different inflammatory myopathies, CD8+ T cells have been expanded directly from muscle and their interactions with autologous myotubes have been investigated in vitro. In several cases, a low but significant autoreactive cytotoxic effect was observed. This is consistent with the hypothesis that some cytotoxic effector T cells recognize an autoantigen on myotubes. One of the major goals for future studies is to define the autoantigens that are relevant in the pathogenesis of the inflammatory myopathies.


The two-process model of sleep regulation: a reappraisal.
In the last three decades the two-process model of sleep regulation has served as a major conceptual framework in sleep research. It has been applied widely in studies on fatigue and performance and to dissect individual differences in sleep regulation. The model posits that a homeostatic process (Process S) interacts with a process controlled by the circadian pacemaker (Process C), with time-courses derived from physiological and behavioural variables. The model simulates successfully the timing and intensity of sleep in diverse experimental protocols. Electrophysiological recordings from the suprachiasmatic nuclei (SCN) suggest that S and C interact continuously. Oscillators outside the SCN that are linked to energy metabolism are evident in SCN-lesioned arrhythmic animals subjected to restricted feeding or methamphetamine administration, as well as in human subjects during internal desynchronization. In intact animals these peripheral oscillators may dissociate from the central pacemaker rhythm. A sleep/fast and wake/feed phase segregate antagonistic anabolic and catabolic metabolic processes in peripheral tissues. A deficiency of Process S was proposed to account for both depressive sleep disturbances and the antidepressant effect of sleep deprivation. The model supported the development of novel non-pharmacological treatment paradigms in psychiatry, based on manipulating circadian phase, sleep and light exposure. In conclusion, the model remains conceptually useful for promoting the integration of sleep and circadian rhythm research. Sleep appears to have not only a short-term, use-dependent function; it also serves to enforce rest and fasting, thereby supporting the optimization of metabolic processes at the appropriate phase of the 24-h cycle.


ALK pERKs up MYCN in neuroblastoma.
The gene expressing the receptor tyrosine kinase anaplastic lymphoma kinase (ALK) is mutated and aberrantly expressed in several cancers. The clinical efficacy of the ALK inhibitor, crizotinib, lags behind expectations for treating MYCN-amplified, ALK-mutant neuroblastoma, a deadly childhood cancer. In this issue of Science Signaling, Umapathy et al. identify the kinase extracellular signal-regulated kinase 5 (ERK5) as a central mediator that enables ALK to boost MYCN expression, and they show that inhibiting ERK5 in concert with ALK reduced neuroblastoma cell viability in vitro and in xenograft tumor models. This report has important clinical implications for the treatment of patients with neuroblastoma or other tumors that overexpress MYC(N) and harbor ALK mutations, such as non-small-cell lung cancer.


Reproducibility of physiological and performance measures from a squash-specific fitness test.
We examined the reproducibility of performance and physiological responses on a squash-specific incremental test. Eight trained squash players habituated to procedures with two prior visits performed an incremental squash test to volitional exhaustion on two occasions 7 days apart. Breath-by-breath oxygen uptake (Vo2) and heart rate were determined continuously using a portable telemetric system. Blood lactate concentration at the end of 4-min stages was assessed to determine lactate threshold. Once threshold was determined, test speed was increased every minute until volitional exhaustion for assessment of maximal oxygen uptake (Vo2max), maximum heart rate (HRmax), and performance time. Economy was taken as the 60-s mean of Vo2 in the final minute of the fourth stage (below lactate threshold for all participants). Typical error of measurement (TEM) with associated 90% confidence intervals, limits of agreement, paired sample t tests, and least products regression were used to assess the reproducibility of scores. Performance time (TEM 27 s, 4%, 90% CI 19 to 49 s) Vo2max (TEM 2.4 mL.kg-1.min-1, 4.7%, 90% CI 1.7 to 4.3 mL.kg-1.min-1), maximum heart rate (TEM 2 beats.min-1, 1.3%, 90% CI 2 to 4 beats.min-1), and economy (TEM 1.6 mL.kg-1.min-1, 4.1%, 90% CI 1.1 to 2.8 mL.kg-1.min-1) were reproducible. The results suggest that endurance performance and physiological responses to a squash-specific fitness test are reproducible.


Clinical Effectiveness of Dry Needling Immediately After Application on Myofascial Trigger Point in Upper Trapezius Muscle.
The purpose of this study was to investigate the effect of dry needling (DN) on pain intensity and pressure pain threshold (PPT) compared with ischemic compression (IC) immediately and 48 hours after each treatment session in individuals with myofascial trigger points in the upper trapezius muscle. Thirty-one patients with myofascial trigger points in the upper trapezius muscle participated in this study. Patients were randomly assigned to a standard (N = 17) or experimental group (N = 14). The treatment protocol for the standard group consisted of IC, whereas the patients in the experimental group received DN. The results indicated that the effect size of the DN methods for pain intensity and PPT was considerably greater after 2 days compared with immediately after the treatment session. In contrast, the effect of the IC for PPT was greater immediately after treatment compared with the measures after 2 days. There was also no noticeable difference in the effect size for IC on pain intensity between the scores obtained immediately and 2 days after treatment. However, our data also revealed a greater effect size for DN on PPT after 2 days compared with the IC technique. In this study, DN improved the pain intensity and PPT after 2 days. However, it had no clinical improvement immediately after application because of muscle soreness. Thus, assessment of the effect of DN immediately after application can be criticized, and the results should be interpreted with caution.


Feasibility and pilot efficacy of cognitive remediation for people in residential substance use treatment.
There are high rates of cognitive impairment among people seeking treatment for problematic substance use, which is rarely addressed in treatment programs. The aim of this pilot study was to evaluate the feasibility and outcomes of the neuropsychological and educational approach to cognitive remediation (NEAR) in a residential substance treatment setting. A non-randomised trial conducted between November 2018 and November 2019, compared outcomes for 34 residents who received cognitive remediation (CR) plus treatment as usual (TAU) to 31 residents who received TAU only. Number of groups and attendance, and a measure of client satisfaction assessed feasibility. Cognitive function was assessed at baseline (service admission), and at 2 and 6 months post-admission. A total of 95 CR groups were delivered over a 36-week time period with a high degree of treatment fidelity and acceptability to participants. Cognitive outcomes improved across both the CR+TAU and TAU groups by 2 months, which was maintained at 6 months for outcomes related to executive functioning. There were no significant differences between the CR+TAU and TAU groups at 2 or 6 months, although a large effect size and confidence intervals indicated a potentially larger change in cognitive flexibility after receiving CR. This study contributes to our understanding of the implementation of CR in the alcohol and other drug rehabilitation setting. It is feasible to incorporate the NEAR program in treatment. The small sample size and lower than expected treatment dose likely contributed to the lack of significant findings.


A neural link between generosity and happiness.
Generous behaviour is known to increase happiness, which could thereby motivate generosity. In this study, we use functional magnetic resonance imaging and a public pledge for future generosity to investigate the brain mechanisms that link generous behaviour with increases in happiness. Participants promised to spend money over the next 4 weeks either on others (experimental group) or on themselves (control group). Here, we report that, compared to controls, participants in the experimental group make more generous choices in an independent decision-making task and show stronger increases in self-reported happiness. Generous decisions engage the temporo-parietal junction (TPJ) in the experimental more than in the control group and differentially modulate the connectivity between TPJ and ventral striatum. Importantly, striatal activity during generous decisions is directly related to changes in happiness. These results demonstrate that top-down control of striatal activity plays a fundamental role in linking commitment-induced generosity with happiness.


Fibroblast growth factor-2 and -4 promote the proliferation of bone marrow mesenchymal stem cells by the activation of the PI3K-Akt and ERK1/2 signaling pathways.
Bone marrow mesenchymal stem cells (BMMSCs) have the capacity for self-renewal, and differentiation into a variety of cell types. They thus represent an attractive source of material for cell therapy. However, little is known about the mechanisms underlying the proliferation of BMMSCs. The purpose of this study was to identify the factors and signaling pathways involved in the proliferation of stem cell antigen-1(+) (Sca-1(+)) BMMSCs. Among the cytokines and growth factors examined in this study, fibroblast growth factor-2 (FGF-2) and FGF-4 significantly stimulated the proliferation of Sca-1(+) BMMSCs, as determined by bromodeoxyuridine incorporation. PI3K-Akt, ERK1/2, and JAK/STAT3 pathways were investigated after stimulation with FGF-2 or FGF-4 via Western blot analysis. No changes were observed in the total ERK1/2 and Akt; however, the pERK1/2 and pAkt levels were upregulated early within 15 min in the FGF-2- or FGF-4-treated Sca-1(+) BMMSCs. Moreover, the pERK1/2 and pAkt upregulation induced by FGF-2 and -4 were completely abolished by treatment with the MEK1/2 inhibitor, U0126 and the PI3K inhibitor, LY294002. However, no change in pJAK2 or total JAK2 levels was observed in the Sca-1(+) BMMSCs induced by FGF-2 or FGF-4. As a consequence of PI3K-Akt and ERK1/2, the upregulation of c-Jun in the Sca-1(+) BMMSCs, after stimulation with FGF-2 or FGF-4, was observed after 12 and 24 h. Moreover, the activation of c-Jun in FGF-2- and FGF-4-treated Sca-1(+) BMMSCs was significantly reduced by U0126. Taken together, these data suggest that FGF-2 and -4 promote the proliferation of Sca-1(+) BMMSCs by activation of the ERK1/2 and PI3K-Akt signaling pathways.


Enhancement of tumor-reactive cytotoxic CD4+ T cell responses after ipilimumab treatment in four advanced melanoma patients.
CD4(+) T cells provide help to enhance and sustain cytotoxic CD8(+) T cell responses. A direct lytic role for this cell population in mouse models further supports the use of tumor-reactive CD4(+) T cells for cancer immunotherapy. CTLA-4 blockade has been shown to expand antigen-specific cytotoxic CD4(+) T cells in mouse models. We took advantage of spontaneous immunity to the NY-ESO-1 cancer-testis antigen to investigate quantitative and qualitative changes in antigen-specific CD4(+) T cell responses after ipilimumab (anti-CTLA-4 monoclonal antibody) treatment in advanced melanoma patients. Four NY-ESO-1 seropositive melanoma patients were chosen upon the availability of suitable blood specimens for characterizing the functions of NY-ESO-1 antigen-specific CD4(+) T cell response by enzyme-linked immunospot (ELISPOT), intracellular cytokine staining (ICS) and cytotoxicity assays. Multiple NY-ESO-1 antigen-specific CD4(+) T cell responses with Th1 dominance were induced or enhanced after ipilimumab treatment in peripheral blood in all four patients. NY-ESO-1 antigen-specific CD4(+) T cell lines established from all 4 patients after ipilimumab treatment recognized naturally processed NY-ESO-1 protein in antigen-presenting cells, expressed master transcription factor Eomesodermin (Eomes) and secreted perforin and Granzyme B. Finally, we demonstrated that these NY-ESO-1 antigen-specific CD4(+) T cell lines directly lysed autologous melanoma cell lines expressing NY-ESO-1 in an MHC class II restricted manner. Our results show that antigen specific cytotoxic CD4(+) T cell responses are induced after ipilimumab therapy in human cancer patients. Ipilimumab may induce the expression of lytic granules on antigen specific cytotoxic CD4(+) T cells via Eomes, revealing a novel consequence of immunologic checkpoint blockade.


Review: Selecting for improved feed efficiency and reduced methane emissions in dairy cattle.
It may be possible for dairy farms to improve profitability and reduce environmental impacts by selecting for higher feed efficiency and lower methane (CH4) emission traits. It remains to be clarified how CH4 emission and feed efficiency traits are related to each other, which will require direct and accurate measurements of both of these traits in large numbers of animals under the conditions in which they are expected to perform. The ranking of animals for feed efficiency and CH4 emission traits can differ depending upon the type and duration of measurement used, the trait definitions and calculations used, the period in lactation examined and the production system, as well as interactions among these factors. Because the correlation values obtained between feed efficiency and CH4 emission data are likely to be biased when either or both are expressed as ratios, therefore researchers would be well advised to maintain weighted components of the ratios in the selection index. Nutrition studies indicate that selecting low emitting animals may result in reduced efficiency of cell wall digestion, that is NDF, a key ruminant characteristic in human food production. Moreover, many interacting biological factors that are not measured directly, including digestion rate, passage rate, the rumen microbiome and rumen fermentation, may influence feed efficiency and CH4 emission. Elucidating these mechanisms may improve dairy farmers ability to select for feed efficiency and reduced CH4 emission.


Maternity care practices: implications for breastfeeding.
Many United States mothers never breastfeed their infants or do so for very short periods. The Baby-Friendly Hospital Initiative was developed to help make breastfeeding the norm in birthing environments, and consists of specific recommendations for maternity care practices. The objective of the current study was to assess the impact of the type and number of Baby-Friendly practices experienced on breastfeeding. A longitudinal mail survey (1993-1994) was administered to women prenatally through 12 months postpartum. The study focused on the 1085 women with prenatal intentions to breastfeed for more than 2 months who initiated breastfeeding, using data from the prenatal and neonatal periods. Predictor variables included indicators of the absence of specific Baby-Friendly practices (late breastfeeding initiation, introduction of supplements, no rooming-in, not breastfeeding on demand, use of pacifiers), and number of Baby-Friendly practices experienced. The main outcome measure was breastfeeding termination before 6 weeks. Only 7 percent of mothers experienced all five Baby-Friendly practices. The strongest risk factors for early breastfeeding termination were late breastfeeding initiation and supplementing the infant. Compared with mothers experiencing all five Baby-Friendly practices, mothers experiencing none were approximately eight times more likely to stop breastfeeding early. Additional practices decreased the risk for early termination. Increased Baby-Friendly Hospital Initiative practices improve the chances of breastfeeding beyond 6 weeks. The need to work with hospitals to increase adoption of these practices is illustrated by the small proportion of mothers who experienced all five practices measured in this study.


Competitive interactions are mediated in a sex-specific manner by arbuscular mycorrhiza in Antennaria dioica.
Plants usually interact with other plants, and the outcome of such interaction ranges from facilitation to competition depending on the identity of the plants, including their sexual expression. Arbuscular mycorrhizal (AM) fungi have been shown to modify competitive interactions in plants. However, few studies have evaluated how AM fungi influence plant intraspecific and interspecific interactions in dioecious species. The competitive abilities of female and male plants of Antennaria dioica were examined in a greenhouse experiment. Females and males were grown in the following competitive settings: (i) without competition, (ii) with intrasexual competition, (iii) with intersexual competition, and (iv) with interspecific competition by Hieracium pilosella - a plant with similar characteristics to A. dioica. Half of the pots were grown with Claroideoglomus claroideum, an AM fungus isolated from the same habitat as the plant material. We evaluated plant survival, growth, flowering phenology, and production of AM fungal structures. Plant survival was unaffected by competition or AM fungi. Competition and the presence of AM fungi reduced plant biomass. However, the sexes responded differently to the interaction between fungal and competition treatments. Both intra- and interspecific competition results were sex-specific, and in general, female performance was reduced by AM colonization. Plant competition or sex did not affect the intraradical structures, extraradical hyphae, or spore production of the AM fungus. These findings suggest that plant sexual differences affect fundamental processes such as competitive ability and symbiotic relationships with AM fungi.


Predictors of loneliness in U.S. adults over age sixty-five.
The purpose of this study was to examine sociodemographic and health-related risks for loneliness among older adults using Health and Retirement Study Data. Overall prevalence of loneliness was 19.3%. Marital status, self-report of health, number of chronic illnesses, gross motor impairment, fine motor impairment, and living alone were predictors of loneliness. Age, female gender, use of home care, and frequency of healthcare visits were not predictive. Loneliness is a prevalent problem for older adults in the United States with its own health-related risks. Future research of interventions targeting identified risks would enhance the evidence base for nursing and the problem of loneliness.


A genetic regulatory network for Xenopus mesendoderm formation.
We have constructed a genetic regulatory network (GRN) summarising the functional relationships between the transcription factors (TFs) and embryonic signals involved in Xenopus mesendoderm formation. It is supported by a relational database containing the experimental evidence and both are available in interactive form via the World Wide Web. This network highlights areas for further study and provides a framework for systematic interrogation of new data. Comparison with the equivalent network for the sea urchin identifies conserved features of the deuterostome ancestral pathway, including positive feedback loops, GATA factors, SoxB, Brachyury and a previously underemphasised role for beta-catenin. In contrast, some features central to one species have not yet been found in the other, for example, Krox and Otx in sea urchin, and Mix and Nodal in Xenopus. Such differences may represent evolved features or may eventually be resolved. For example, in Xenopus, Nodal-related genes are positively regulated by beta-catenin and at least one of them is repressed by Sox3, as is the uncharacterised early signal (ES) inducing endomesoderm in the sea urchin, suggesting that ES may be a Nodal-like TGF-beta. Wider comparisons of such networks will inform our understanding of developmental evolution.


Reducing crashes and injuries among young drivers: what kind of prevention should we be focusing on?
Every year, drivers throughout the world are killed or injured in road traffic. Young drivers run a greater risk everywhere, and this problem is still largely unsolved. Better understanding of the underlying processes could, however, be a useful tool in preventive endeavours. To change a young driver's goals behind driving and the context in which it is done, a variety of different methods of persuasion should be tested. Both "soft" and "hard" methods should be used. For example, communication and increased enforcement may be used simultaneously. Communication campaigns should highlight the dangers of unsafe behaviour and in particular target young males. Communication campaigns that employ persuasive, emotional messages are most effective where young drivers are concerned. Research shows that attitudes about safety are formed at an early age, long before legal driving, and therefore it would also be important to target young adolescents. Laws need enforcement to be effective and should target areas of particular risk to young drivers. Driver education or communication campaigns cannot be expected to radically change a young person's life goals. For that purpose, active learning methods that make use of the learner's own experiences have to be applied. Special courses for young drivers designed to make individuals conscious of their personal tendencies and the type of social context that affects their driving behaviour could be helpful, whether offered via the ordinary school system or at driving schools.


A two-player iterated survival game.
We describe an iterated game between two players, in which the payoff is to survive a number of steps. Expected payoffs are probabilities of survival. A key feature of the game is that individuals have to survive on their own if their partner dies. We consider individuals with hardwired, unconditional behaviors or strategies. When both players are present, each step is a symmetric two-player game. The overall survival of the two individuals forms a Markov chain. As the number of iterations tends to infinity, all probabilities of survival decrease to zero. We obtain general, analytical results for n-step payoffs and use these to describe how the game changes as n increases. In order to predict changes in the frequency of a cooperative strategy over time, we embed the survival game in three different models of a large, well-mixed population. Two of these models are deterministic and one is stochastic. Offspring receive their parent's type without modification and fitnesses are determined by the game. Increasing the number of iterations changes the prospects for cooperation. All models become neutral in the limit (n→∞). Further, if pairs of cooperative individuals survive together with high probability, specifically higher than for any other pair and for either type when it is alone, then cooperation becomes favored if the number of iterations is large enough. This holds regardless of the structure of pairwise interactions in a single step. Even if the single-step interaction is a Prisoner's Dilemma, the cooperative type becomes favored. Enhanced survival is crucial in these iterated evolutionary games: if players in pairs start the game with a fitness deficit relative to lone individuals, the prospects for cooperation can become even worse than in the case of a single-step game.


Selenium- and Tellurium-Based Antioxidants for Modulating Inflammation and Effects on Osteoblastic Activity.
Increased oxidative stress plays a significant role in the etiology of bone diseases. Heightened levels of H2O2 disrupt bone homeostasis, leading to greater bone resorption than bone formation. Organochalcogen compounds could act as free radical trapping agents or glutathione peroxidase mimetics, reducing oxidative stress in inflammatory diseases. In this report, we synthesized and screened a library of organoselenium and organotellurium compounds for hydrogen peroxide scavenging activity, using macrophagic cell lines RAW264.7 and THP-1, as well as human mono- and poly-nuclear cells. These cells were stimulated to release H2O2, using phorbol 12-myristate 13-acetate, with and without organochalogens. Released H2O2 was then measured using a chemiluminescent assay over a period of 2 h. The screening identified an organoselenium compound which scavenged H2O2 more effectively than the vitamin E analog, Trolox. We also found that this organoselenium compound protected MC3T3 cells against H2O2-induced toxicity, whereas Trolox did not. The organoselenium compound exhibited no cytotoxicity to the cells and had no deleterious effects on cell proliferation, viability, or alkaline phosphatase activity. The rapidity of H2O2 scavenging and protection suggests that the mechanism of protection is due to the direct scavenging of extracellular H2O2. This compound is a promising modulators of inflammation and could potentially treat diseases involving high levels of oxidative stress.


Computational markers of experience- but not description-based decision-making are associated with future depressive symptoms in young adults.
Early prediction of high depressive symptoms is crucial for selective intervention and the minimization of functional impairment. Recent cross-sectional studies indicated decision-making deficits in depression, which may be an important contributor to the disorder. Our goal was to test whether description- and experience-based decision making, two major neuroeconomic paradigms of decision-making under uncertainty, predict future depressive symptoms in young adults. One hundred young adults performed two decision-making tasks, one description-based, in which subjects chose between two gambling options given explicitly stated rewards and their probabilities, and the other experience-based, in which subjects were shown rewards but had to learn the probability of those rewards (or cue-outcome contingencies) via trial-and-error experience. We evaluated subjects' depressive symptoms with BDI-II at baseline (T1) and half a year later (T2). Comparing subjects with low versus high levels of depressive symptoms at T2 showed that the latter performed worse on the experience- but not description-based task at T1. Computational modeling of the decision-making process suggested that subjects with high levels of depressive symptoms had a more concave utility function, indicating enhanced risk aversion. Furthermore, a more concave utility function at T1 increased the odds of high depressive symptoms at T2, even after controlling depressive symptoms at T1, perceived stress at T2, and several covariates (OR = 0.251, 95% CI [0.085, 0.741]). This is the first study to demonstrate a prospective link between experience-based decision-making and depressive symptoms. Our results suggest that enhanced risk aversion in experience-based decision-making may be an important contributor to the development of depressive symptoms.


Allogeneic hematopoietic stem cell transplantation with reduced-intensity conditioning in acute lymphoblastic leukemia: a feasibility study.
Numerous studies have reported the feasibility of performing allogeneic hematopoietic stem cell transplantation with reduced-intensity conditioning, although results in individual diseases are scarce, with no studies in patients with acute lymphoblastic leukemia (ALL). We sought to analyze the results of reduced intensity conditioning for allografts in adult patients with ALL. We report the results of a reduced intensity conditioning regimen followed by allogeneic hematopoietic stem cell transplantation in 27 adult patients with high-risk ALL who were included in four prospective studies. The median age was 50 years; 23 (85%) patients were beyond first complete remission, 44% were chemorefractory and 41% were Philadelphia chromosome positive. Donors were mismatched related donors or volunteer unrelated donors in 12 cases (44%). The incidence of grades II-IV acute graft-versus-host disease (GVHD) was 48%, and 13 of 18 evaluable patients (72%) developed chronic GVHD. Currently nine patients are alive, with a median follow-up of 809 days (range, 381-1375). The 2-year incidence of transplant-related mortality was 23% (95% CI, 11% to 46%), and the 2-year probability of overall survival was 31% (95% CI, 12 to 48%), while the 2-year incidence of disease progression was 49% (95% CI, 33% to 72%). The 2-year incidence of disease progression in patients with and without GVHD was 35% (95% CI, 19% to 57%) and 70% (95% CI, 47% to 100%), respectively (p=0.05). This retrospective study suggests that allogeneic hematopoietic stem cell transplantation with reduced intensity conditioning might be a useful therapeutic option for some patients with ALL who are ineligible for standard myeloablative conditioning. However, this treatment modality needs to be evaluated in prospective trials, and should not be employed outside clinical studies.


Prospective associations between problematic eating attitudes in midchildhood and the future onset of adolescent obesity and high blood pressure.
Clinically diagnosed eating disorders may have adverse cardiometabolic consequences, including overweight or obesity and high blood pressure. However, the link between problematic eating attitudes in early adolescence, which can lead to disordered eating behaviors, and future cardiometabolic health is, to our knowledge, unknown. We assessed whether variations in midchildhood eating attitudes influence the future development of overweight or obesity and high blood pressure. Of 17,046 children who participated in the Promotion of Breastfeeding Intervention Trial (PROBIT), we included 13,557 participants (79.5% response rate) who completed the Children's Eating Attitudes Test (ChEAT) at age 11.5 y and in whom we measured adiposity and blood pressure at ages 6.5, 11.5, and 16 y. We assessed whether ChEAT scores ≥85th percentile (indicative of problematic eating attitudes) compared with scores <85th percentile at age 11.5 y were associated with new-onset overweight, obesity, high systolic blood pressure, or high diastolic blood pressure between midchildhood and early adolescence. After controlling for baseline sociodemographic confounders, we observed positive associations of problematic eating attitudes at age 11.5 y with new-onset obesity (OR: 2.18; 95% CI: 1.58, 3.02), new-onset high systolic blood pressure (OR: 1.34; 95% CI: 1.05, 1.70), and new-onset high diastolic blood pressure (OR: 1.25; 95% CI: 0.99, 1.58) at age 16 y. After further controlling for body mass index at age 6.5 y, problematic eating attitudes remained positively associated with new-onset obesity (OR: 1.80; 95% CI: 1.28, 2.53); however, associations with new-onset high blood pressure were attenuated (OR: 1.14; 95% CI: 0.89, 1.45 and OR: 1.09; 95% CI: 0.86, 1.39 for new-onset systolic and diastolic blood pressure, respectively). Problematic eating attitudes in midchildhood seem to be related to the development of obesity in adolescence, a relatively novel observation with potentially important public health implications for obesity control. PROBIT was registered at clinicaltrials.gov as NCT01561612 and isrctn.com as ISRCTN37687716.


Thioredoxin-interacting protein, hematopoietic stem cells, and hematopoiesis.
Reactive oxygen species (ROS) can regulate diverse signaling pathways and functions in hematopoietic cells. Thioredoxin-interacting protein (TXNIP) plays an important role in mammalian cells by inhibiting thioredoxin (TRX) under oxidative stress conditions. TXNIP is expressed in hematopoietic stem cells (HSCs), and its expression decreases as HSCs differentiate into precursor cells. However, this reduction in expression does not sufficiently explain the function of TXNIP in hematopoietic cells under oxidative stress conditions. Here, we review how ROS can regulate hematopoiesis by focusing on the function of TXNIP in hematopoietic cells under oxidative stress conditions. Studies of Txnip mice have demonstrated an antioxidant function of TXNIP in hematopoietic cells or immune cells. This antioxidant function differs from the conventional pro-oxidant activity of TXNIP observed in other cell types under oxidative stress. The data suggest a context-dependent function of TXNIP under oxidative stress conditions and, in particular, a differential function of TXNIP in hematopoietic cells via its direct interaction with other redox regulatory proteins. The regulation of ROS is important in determining cellular fate decisions. TXNIP acts as a negative regulator of TRX via direct interaction, and it increases the levels of ROS under oxidative stress. However, TXNIP has an antioxidant function in hematopoietic cells or immune cells, as ROS levels are elevated and induce apoptosis in Txnip hematopoietic cells. These results suggest that the amount of TXNIP is inversely associated with ROS levels, and the loss of TXNIP can increase ROS levels in immune cells or hematopoietic cells.


Pathological features of COVID-19 infection from biopsy and autopsy series.
Novel coronavirus disease 2019 (COVID-19) which is caused by severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) was first identified in December 2019 in Chinese town Wuhan and considered as a pandemic by World Health Organization. The disease has variety of symptoms including fever, shortness of breath, cough, fatigue, loss of smell and taste and diarrhea. While the majority of cases have mild symptoms, some progress to viral pneumonia, multi-organ failure, or cytokine storm and mortality is mostly caused by hypoxemic respiratory failure. Until now, more than 3.5 million people worldwide were infected and more than 240.000 mortality has been occurred. Thus, there is now evidence the disease may affect variety of organs according to accumulating biopsy and autopsy studies. Such pathological studies have potential role on the understanding of clinical outcomes and in the development of novel targeted therapeutic approaches. Given these aforementioned data, in the current manuscript we have summarized the pathological features of COVID-19 derived from biopsy and autopsy series.


[Functional dependence and chronic disease in older Mexicans].
The aim of this study is to determine the prevalence of functional dependence in older Mexicans and associated chronic conditions. The study was conducted between June and December 2004 in the Geriatric Department of the Instituto Nacional de Ciencias Medicas y Nutrición Salvador Zubiran, Mexico City. The data for the variables were obtained from the information in the National Study of Health and Aging in Mexico survey (ENASEM). The survey was based on the law for statistical and geographic information,Chapter V, Article 38. A total of 4,872 subjects from the national survey of aging and health in Mexico were asked about their need for help in basic activities of daily living (BADL) and instrumental activities of daily living (IADL). Presence of disease was obtained through self-reporting. Prevalence of functional dependence was 24 and 23% for BADL and IADL, respectively. Joint disease, cerebrovascular disease, cognitive impairment and depression were associated with dependency in BADL and IADL. The prevalence of functional dependence is high among Mexicans over 65 years old. This study shows the chronic conditions that are closely associated with functional dependence and provides a perspective in order to develop preventative measures.


Role of the copper transporter, CTR1, in platinum-induced ototoxicity.
The goal of this study was to determine the role of an influx copper transporter, CTR1, in the ototoxicity induced by cisplatin, a potent anticancer platinum analog used in the treatment of a variety of solid tumors. As determined through reverse transcriptase-PCR (RT-PCR), quantitative RT-PCR, Western blot, and immunohistochemistry, mouse CTR1 (Ctr1) was found to be abundantly expressed and highly localized at the primary sites of cisplatin toxicity in the inner ear, mainly outer hair cells (OHCs), inner hair cells, stria vascularis, spiral ganglia, and surrounding nerves in the mouse cochlea. A CTR1 substrate, copper sulfate, decreased the uptake and cytotoxicity of cisplatin in HEI-OC1, a cell line that expresses many molecular markers reminiscent of OHCs. Small interfering RNA-mediated knockdown of Ctr1 in this cell line caused a corresponding decrease in cisplatin uptake. In mice, intratympanic administration of copper sulfate 30 min before intraperitoneal administration of cisplatin was found to prevent hearing loss at click stimulus and 8, 16, and 32 kHz frequencies. To date, the utility of cisplatin remains severely limited because of its ototoxic effects. The studies described in this report suggest that cisplatin-induced ototoxicity and cochlear uptake can be modulated by administration of a CTR1 inhibitor, copper sulfate. The possibility of local administration of CTR1 inhibitors during cisplatin therapy as a means of otoprotection is thereby raised.


Staffing challenges and strategies for organizations serving individuals who have experienced chronic homelessness.
Hiring and retaining appropriate staff is essential for programs serving those who have experienced chronic homelessness. This paper describes specific staffing challenges and strategies from the Collaborative Initiative to Help End Chronic Homelessness (CICH), an 11-site, multi-agency Federal program designed to serve people experiencing chronic homelessness who also have a disabling condition such as substance use or mental health problems. This paper addresses approaches to staffing including team structures, staff supervision, and training. Challenges identified include low pay, high rates of burnout and turnover, limited time for supervision, and multiple staff training needs. This paper also explores specific staffing strategies based on the experience of the CICH sites, and concludes with implications for practice, research, and policy, including recommendations for ongoing staff training, suggestions for future mixed-methods research, and a call for an enhanced focus on strengthening the homeless services workforce.


Mycobacterium immunogenum sp. nov., a novel species related to Mycobacterium abscessus and associated with clinical disease, pseudo-outbreaks and contaminated metalworking fluids: an international cooperative study on mycobacterial taxonomy.
PCR-restriction enzyme pattern analysis of a 439 bp hsp65 gene segment identified 113 unique isolates among non-pigmented rapidly growing mycobacteria (RGM) from clinical and environmental sources that failed to match currently recognized species patterns. This group represented 40% of isolates recovered from bronchoscope contamination pseudo-outbreaks, 0% of disease-associated nosocomial outbreaks and 4% of routine clinical isolates of the Mycobacterium abscessus/Mycobacterium chelonae group submitted to the Mycobacteria/Nocardia laboratory for identification. It is grouped within the Mycobacterium fortuitum complex, with growth in less than 7 d, absence of pigmentation, positive 3-d arylsulfatase reaction and growth on MacConkey agar without crystal violet. It exhibited overlapping biochemical, antimicrobial susceptibility and HPLC characteristics of M. abscessus and M. chelonae. By 16S rRNA gene sequencing, these isolates comprised a homogeneous group with a unique hypervariable region A sequence and differed by 8 and 10 bp, respectively, from M. abscessus and M. chelonae. Surprisingly, this taxon contained two copies of the ribosomal operon, compared with single copies in the two related species. By DNA-DNA hybridization, this new group exhibited <30% homology with recognized RGM species. The name Mycobacterium immunogenum sp. nov. is proposed for this new taxon.


Freeze-dried plasma at the point of injury: from concept to doctrine.
While early plasma transfusion for the treatment of patients with ongoing major hemorrhage is widely accepted as part of the standard of care in the hospital setting, logistic constraints have limited its use in the out-of-hospital setting. Freeze-dried plasma (FDP), which can be stored at ambient temperatures, enables early treatment in the out-of-hospital setting. Point-of-injury plasma transfusion entails several significant advantages over currently used resuscitation fluids, including the avoidance of dilutional coagulopathy, by minimizing the need for crystalloid infusion, beneficial effects on endothelial function, physiological pH level, and better maintenance of intravascular volume compared with crystalloid-based solutions. The Israel Defense Forces Medical Corps policy is that plasma is the resuscitation fluid of choice for selected, severely wounded patients and has thus included FDP as part of its armamentarium for use at the point of injury by advanced life savers, across the entire military. We describe the clinical rationale behind the use of FDP at the point-of-injury, the drafting of the administration protocol now being used by Israel Defense Forces advanced life support providers, the process of procurement and distribution, and preliminary data describing the first casualties treated with FDP at the point of injury. It is our hope that others will be able to learn from our experience, thus improving trauma casualty care around the world.


Bond strength of composite resin to pulp capping biomaterials after application of three different bonding systems.
Background and aims. Bonding of composite resin filling materials to pulp protecting agents produces an adhesive joint which is important for the quality of filling as well as success of restoration. We aimed to assess the bond strength of composite resin to three pulp capping biomaterials: Pro Root mineral trioxide aggregate (PMTA), Root MTA (RMTA) and calcium enriched mixture (CEM) cement, using three bonding systems [a total-etch (Single Bond) and two self-etch systems (Protect bond and SE Bond)]. Materials and methods. Ninety acrylic molds, each containing a 6×2-mm hole, were divided into 3 groups and filled with PMTA, RMTA and CEM cements. The samples in each experimental group were then randomly divided into 3 sub-groups; Single Bond, Protect Bond and SE Bond bonding systems were applied to the tested materials. Cylindrical forms of composite resin (Z100, 2×2 mm) were placed onto the samples and cured. Shear bond strength values were measured for 9 subgroups using a universal testing machine. Data were analyzed using two-way ANOVA. Results. The average shear bond strengths of Z100 composite resin after application of Single Bond, Protect Bond and SE Bond systems were as follows; PMTA: 5.1±2.42, 4.56±1.96 and 4.52±1.7; RMTA: 4.71±1.77, 4.31±0.56 and 4.79±1.88; and CEM cement: 4.75±1.1, 4.54±1.59 and 4.64±1.78 MPa, respectively. The type of pulp capping material, bonding system and their interacting effects did not have a significant effect on the bond strengths of composite resin to pulp capping biomaterials. Conclusion. Within the limitations of this in vitrostudy, bond strength of composite resin to two types of MTA as well as CEM cement were similar following application of the total-etch or self-etch bonding systems.


The consistent differential expression of genetic pathways following exposure of an industrial Pseudomonas aeruginosa strain to preservatives and a laundry detergent formulation.
Pseudomonas aeruginosa is a common contaminant associated with product recalls in the home and personal care industry. Preservation systems are used to prevent spoilage and protect consumers, but greater knowledge is needed of preservative resistance mechanisms used by P. aeruginosa contaminants. We aimed to identify genetic pathways associated with preservative exposure by using an industrial P. aeruginosa strain and implementing RNA-Seq to understand gene expression changes in response to industry relevant conditions. The consistent differential expression of five genetic pathways during exposure to multiple industrial growth conditions associated with benzisothiazolone (BIT) and phenoxyethanol (POE) preservatives, and a laundry detergent (LD) formulation, was observed. A MexPQ-OpmE Resistance Nodulation Division efflux pump system was commonly upregulated in response to POE, a combination of BIT and POE, and LD together with BIT. In response to all industry conditions, a putative sialic acid transporter and isoprenoid biosynthesis gnyRDBHAL operon demonstrated consistent upregulation. Two operons phnBA and pqsEDCBA involved in Pseudomonas quinolone signaling production and quorum-sensing were also consistently downregulated during exposure to all the industry conditions. The ability to identify consistently differentially expressed genetic pathways in P. aeruginosa can inform the development of future targeted preservation systems that maintain product safety and minimise resistance development.


Ultrasound and microbubble-targeted delivery of macromolecules is regulated by induction of endocytosis and pore formation.
Contrast microbubbles in combination with ultrasound (US) are promising vehicles for local drug and gene delivery. However, the exact mechanisms behind intracellular delivery of therapeutic compounds remain to be resolved. We hypothesized that endocytosis and pore formation are involved during US and microbubble targeted delivery (UMTD) of therapeutic compounds. Therefore, primary endothelial cells were subjected to UMTD of fluorescent dextrans (4.4 to 500 kDa) using 1 MHz pulsed US with 0.22-MPa peak-negative pressure, during 30 seconds. Fluorescence microscopy showed homogeneous distribution of 4.4- and 70-kDa dextrans through the cytosol, and localization of 155- and 500-kDa dextrans in distinct vesicles after UMTD. After ATP depletion, reduced uptake of 4.4-kDa dextran and no uptake of 500-kDa dextran was observed after UMTD. Independently inhibiting clathrin- and caveolae-mediated endocytosis, as well as macropinocytosis significantly decreased intracellular delivery of 4.4- to 500-kDa dextrans. Furthermore, 3D fluorescence microscopy demonstrated dextran vesicles (500 kDa) to colocalize with caveolin-1 and especially clathrin. Finally, after UMTD of dextran (500 kDa) into rat femoral artery endothelium in vivo, dextran molecules were again localized in vesicles that partially colocalized with caveolin-1 and clathrin. Together, these data indicated uptake of molecules via endocytosis after UMTD. In addition to triggering endocytosis, UMTD also evoked transient pore formation, as demonstrated by the influx of calcium ions and cellular release of preloaded dextrans after US and microbubble exposure. In conclusion, these data demonstrate that endocytosis is a key mechanism in UMTD besides transient pore formation, with the contribution of endocytosis being dependent on molecular size.


Becoming a medical educator: motivation, socialisation and navigation.
Despite an increasing concern about a future shortage of medical educators, little published research exists on career choices in medical education nor the impact of specific training posts in medical education (e.g. academic registrar/resident positions). Medical educators at all levels, from both medical and non-medical backgrounds, are crucial for the training of medical students, junior doctors and in continuing professional development. We explored the motivations and experiences of junior doctors considering an education career and undertaking a medical education registrar (MER) post. Data were collected through semi-structured interviews with junior doctors and clinicians across Queensland Health. Framework analysis was used to identify themes in the data, based on our defined research questions and the medical education workforce issues prompting the study. We applied socio-cognitive career theory to guide our analysis and to explore the experience of junior doctors in medical education registrar posts as they enter, navigate and fulfil the role. We identified six key themes in the data: motivation for career choice and wanting to provide better education; personal goals, expectations and the need for self-direction; the influence of role models; defining one's identity; support networks and the need for research as a potential barrier to pursuing a career in/with education. We also identified the similarities and differences between the MERs' experiences to develop a composite of an MER's journey through career choice, experience in role and outcomes. There is growing interest from junior doctors in pursuing education pathways in a clinical environment. They want to enhance clinical teaching in the hospitals and become specialists with an interest in education, and have no particular interest in research or academia. This has implications for the recruitment and training of the next generation of clinical educators.


Pro-metastatic splicing of Ron proto-oncogene mRNA can be reversed: therapeutic potential of bifunctional oligonucleotides and indole derivatives.
Alternative splicing is a key molecular mechanism for increasing the complexity of the human transcriptome. Nearly all human genes are regulated by alternative splicing and the deregulation of this process has a causative role in various human diseases, including cancer. The discovery that alternatively spliced isoforms of several genes are expressed selectively in tumor cells opened the exciting possibility that pharmacological treatment of aberrant splicing could lead to new anti-cancer therapeutic approaches. An alternatively spliced isoform of a scatter factor receptor and proto-oncogene, Ron, accumulates during tumor progression of epithelial tissues and is able to confer an invasive phenotype to the expressing cells. This isoform, called ΔRon, originates from skipping of exon 11, and this specific splicing event is controlled by the expression level of the splicing factor and proto-oncogene SF2/ASF. Over-expression of SF2/ASF, which occurs frequently in various human tumors, induces the production of ΔRon and activates the epithelial to mesenchymal transition (EMT), leading to increased cell motility. In this paper, we have used targeted oligonucleotide enhancers of splicing (TOES) to recruit positive splicing factors to Ron exon 11 and thereby stimulate its inclusion. As an alternative approach, we have used selected indole derivatives that target ASF/SF2 splicing activity. Both treatments correct aberrant ΔRon splicing, restoring the incorporation of Ron exon 11. Notably, indole derivatives are also able to affect the invasive phenotype of the cells. Thus, these treatments may have therapeutic applications for anti-cancer purposes.


Participant perceptions and experiences of a novel community-based respiratory longitudinal sampling method in Liverpool, UK: A mixed methods feasibility study.
Longitudinal, community-based sampling is important for understanding prevalence and transmission of respiratory pathogens. Using a minimally invasive sampling method, the FAMILY Micro study monitored the oral, nasal and hand microbiota of families for 6 months. Here, we explore participant experiences and opinions. A mixed methods approach was utilised. A quantitative questionnaire was completed after every sampling timepoint to report levels of discomfort and pain, as well as time taken to collect samples. Participants were also invited to discuss their experiences in a qualitative structured exit interview. We received questionnaires from 36 families. Most adults and children >5y experienced no pain (94% and 70%) and little discomfort (73% and 47% no discomfort) regardless of sample type, whereas children ≤5y experienced variable levels of pain and discomfort (48% no pain but 14% hurts even more, whole lot or worst; 38% no discomfort but 33% moderate, severe, or extreme discomfort). The time taken for saliva and hand sampling decreased over the study. We conducted interviews with 24 families. Families found the sampling method straightforward, and adults and children >5y preferred nasal sampling using a synthetic absorptive matrix over nasopharyngeal swabs. It remained challenging for families to fit sampling into their busy schedules. Adequate fridge/freezer space and regular sample pick-ups were found to be important factors for feasibility. Messaging apps proved extremely effective for engaging with participants. Our findings provide key information to inform the design of future studies, specifically that self-sampling at home using minimally invasive procedures is feasible in a family context.


The effect of cooking process on the total lipid and n-3 LC-PUFA contents of Australian Bass Strait scallops, Pecten fumatus.
A total of twenty-three Australian Bass Strait scallops, Pecten fumatus processed by three different cooking methods: steam, battered and deep-fry, and pan-fry were analysed to determine the total lipid and health-benefiting n-3 PUFA contents. Fry process resulted in a significantly higher lipid content (p<0.05) with 1.98g/100g being found in deep-fried and 1.78g/100g in pan-fried scallops while 1.31g/100g was recorded in the fresh control group (p<0.05). A higher concentration of alpha-linolenic acid (ALA, 18:3n-3), total n-6 PUFA and linoleic (LA, 18:2n-6) were also observed in fried scallops (p<0.05). The two main n-3 LC-PUFA were eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) and their concentration also varied depending on the cooking methods. Pan-fried scallops showed a higher concentration of EPA, DHA and total n-3 PUFA while steamed scallops had a higher concentration of DHA only (p<0.05). The ratio of n-3/n-6 PUFA was markedly lower in both fried groups than in control and steamed scallops (p<0.0001). Three frozen samples were also examined and no significant variations in the lipid profiles have been observed after 22 days. The variation of lipid profiles reflects mainly the uptake of vegetable oil components and loss of water during cooking process. Scallops represent a good source of n-3 LC-PUFA with the concentration ranging from 312.4 mg/100g in fresh scallops to 522.1mg/100g in pan-fried scallops.


Metabolism at the Max: How Vertebrate Organisms Respond to Physical Activity.
Activity metabolism is supported by phosphorylated reserves (adenosine triphosphate, creatine phosphate), glycolytic, and aerobic metabolism. Because there is no apparent variation between vertebrate groups in phosphorylated reserves or glycolytic potential of skeletal muscle, variation in maximal metabolic rate between major vertebrate groups represents selection operating on aerobic mechanisms. Maximal rates of oxygen consumption in vertebrates are supported by increased conductive and diffusive fluxes of oxygen from the environment to the mitochondria. Maximal CO2 efflux from the mitochondria to the environment must be matched to oxygen flux, or imbalances in pH will occur. Among vertebrates, there are a variety of modes of locomotion and vastly different rates of metabolism supported by a variety of cardiorespiratory architectures. However, interclass comparisons strongly implicate systemic oxygen transport as the rate-limiting step to maximal oxygen consumption for all vertebrate groups. The key evolutionary step that accounts for the approximately 10-fold increase in maximal oxygen flux in endotherms versus ectotherms appears to be maximal heart rate. Other variables such as ventilation, pulmonary/gill, and tissue diffusing capacity, have excess capacity and thus are not limiting to maximal oxygen consumption. During maximal activity, the ratio of ventilation to respiratory system blood flow is remarkably similar among vertebrates, and CO2 extraction efficiency increases while oxygen extraction efficiency decreases, suggesting that the respiratory system provides the largest resistance to maximal CO2 flux. Despite the large variation in modes of activity and rates of metabolism, maximal rates of oxygen and CO2 flux appear to be limited by the cardiovascular and respiratory systems, respectively.


DNA ligase III is critical for mtDNA integrity but not Xrcc1-mediated nuclear DNA repair.
DNA replication and repair in mammalian cells involves three distinct DNA ligases: ligase I (Lig1), ligase III (Lig3) and ligase IV (Lig4). Lig3 is considered a key ligase during base excision repair because its stability depends upon its nuclear binding partner Xrcc1, a critical factor for this DNA repair pathway. Lig3 is also present in the mitochondria, where its role in mitochondrial DNA (mtDNA) maintenance is independent of Xrcc1 (ref. 4). However, the biological role of Lig3 is unclear as inactivation of murine Lig3 results in early embryonic lethality. Here we report that Lig3 is essential for mtDNA integrity but dispensable for nuclear DNA repair. Inactivation of Lig3 in the mouse nervous system resulted in mtDNA loss leading to profound mitochondrial dysfunction, disruption of cellular homeostasis and incapacitating ataxia. Similarly, inactivation of Lig3 in cardiac muscle resulted in mitochondrial dysfunction and defective heart-pump function leading to heart failure. However, Lig3 inactivation did not result in nuclear DNA repair deficiency, indicating essential DNA repair functions of Xrcc1 can occur in the absence of Lig3. Instead, we found that Lig1 was critical for DNA repair, but acted in a cooperative manner with Lig3. Additionally, Lig3 deficiency did not recapitulate the hallmark features of neural Xrcc1 inactivation such as DNA damage-induced cerebellar interneuron loss, further underscoring functional separation of these DNA repair factors. Therefore, our data reveal that the critical biological role of Lig3 is to maintain mtDNA integrity and not Xrcc1-dependent DNA repair.


Visual performance after reduced blinking in eyes with soft contact lenses or after LASIK.
To evaluate visual performance during concentrated visual work in patients wearing soft contact lenses or after LASIK. Thirty-one eyes of 17 patients who had worn soft contact lenses before LASIK were examined by the following tests immediately and 10 seconds after eye opening: 1) functional visual acuity, which is defined as visual acuity measured after prolonged eye opening without blinking; 2) surface regularity index (SRI) and surface asymmetry index (SAI) in corneal topography; and 3) higher order aberration measured with NIDEK OPD-Scan. Results were compared in the same patients before (with soft contact lenses and no eye surgery) and 1 month after LASIK (without soft contact lenses). Functional visual acuity was significantly decreased 10 seconds after eye opening compared with immediately after eye opening, both with soft contact lenses and after LASIK, and decreased to a greater extent with soft contact lenses. The SRI and SAI were significantly increased 10 seconds after eye opening compared with immediately after eye opening, both with soft contact lenses and after LASIK, and increased to a greater extent with soft contact lenses. Higher order aberration was increased 10 seconds after eye opening with soft contact lenses, but not after LASIK. Our results suggest that prolonged eye opening induces a decreased quality of vision in eyes of soft contact lens wearers and after LASIK. Under conditions in which blinking is restricted due to concentrated visual work, such as visual display terminal work, reading, and driving, visual performance may be more compromised with soft contact lens wear than after LASIK.


Sexual imprinting on ecologically divergent traits leads to sexual isolation in sticklebacks.
During sexual imprinting, offspring learn parental phenotypes and then select mates who are similar to their parents. Imprinting has been thought to contribute to the process of speciation in only a few rare cases; this is despite imprinting's potential to generate assortative mating and solve the problem of recombination in ecological speciation. If offspring imprint on parental traits under divergent selection, these traits will then be involved in both adaptation and mate preference. Such 'magic traits' easily generate sexual isolation and facilitate speciation. In this study, we show that imprinting occurs in two ecologically divergent stickleback species (benthics and limnetics: Gasterosteus spp.). Cross-fostered females preferred mates of their foster father's species. Furthermore, imprinting is essential for sexual isolation between species; isolation was reduced when females were raised without fathers. Daughters imprinted on father odour and colour during a critical period early in development. These traits have diverged between the species owing to differences in ecology. Therefore, we provide the first evidence that imprinting links ecological adaptation to sexual isolation between species. Our results suggest that imprinting may facilitate the evolution of sexual isolation during ecological speciation, may be especially important in cases of rapid diversification, and thus play an integral role in the generation of biodiversity.


Clinical effects of vinorelbine administration in the management of various malignant tumor types in dogs: 58 cases (1997-2012).
To evaluate the effectiveness of vinorelbine in the management of various malignant tumor types in dogs. Retrospective case series. 58 dogs with malignant tumors, including pulmonary carcinoma (n = 31), histiocytic sarcoma (9), mast cell tumor (5), lymphoma (4), melanoma (2), and 7 other tumor types (1 each). Medical records of dogs treated with vinorelbine from December 1997 to December 2012 were reviewed for data regarding signalment, clinical signs, physical examination findings, clinicopathologic test results, diagnostic imaging results, vinorelbine doses and dose frequency, surgery and radiotherapy details when applicable, other chemotherapeutics administered, and outcomes. Descriptive, comparative, and survival statistics were computed for all dogs and for dogs by histologic subgroup of tumors. Vinorelbine was administered palliatively to 44 (76%) dogs. One (2%) dog had a complete response for 162 days, 5 (11%) dogs had a partial response for a median duration of 91 days, 19 (43%) dogs had stable disease for a median duration of 68 days, and 19 (43%) dogs developed progressive disease after a median duration of 21 days. Clinical benefit was more difficult to assess in the remaining 14 (24%) dogs that received vinorelbine as an adjuvant treatment. Overall median time to tumor progression was 103 days (range, 5 to 1,533 days). Vinorelbine appeared to be effective in the treatment of several tumor types in dogs. Follow-up prospective studies of the clinical benefit of the drug in specific clinical scenarios will be necessary to support this conclusion.


Peptide and nucleic acid-directed self-assembly of cationic nanovehicles through giant unilamellar vesicle modification: Targetable nanocomplexes for in vivo nucleic acid delivery.
One of the greatest challenges for the development of genetic therapies is the efficient targeted delivery of therapeutic nucleic acids. Towards this goal, we have introduced a new engineering initiative in self-assembly of biologically safe and stable nanovesicle complexes (∼90 to 140nm) derived from giant unilamellar vesicle (GUV) precursors and comprising plasmid DNA or siRNA and targeting peptide ligands. The biological performance of the engineered nanovesicle complexes were studied both in vitro and in vivo and compared with cationic liposome-based lipopolyplexes. Compared with cationic lipopolyplexes, nanovesicle complexes did not show advantages in transfection and cell uptake. However, nanovesicle complexes neither displayed significant cytotoxicity nor activated the complement system, which are advantageous for intravenous injection and tumour therapy. On intravenous administration into a neuroblastoma xenograft mouse model, nanovesicle complexes were found to distribute throughout the tumour interstitium, thus providing an alternative safer approach for future development of tumour-specific therapeutic nucleic acid interventions. On oropharyngeal instillation, nanovesicle complexes displayed better transfection efficiency than cationic lipopolyplexes. The technological advantages of nanovesicle complexes, originating from GUVs, over traditional cationic liposome-based lipopolyplexes are discussed. The efficient targeted delivery of nucleic acids in vivo provides some of the greatest challenges to the development of genetic therapies. Giant unilamellar lipid vesicles (GUVs) have been used mainly as cell and tissue mimics and are instrumental in studying lipid bilayers and interactions. Here, the GUVs have been modified into smaller nanovesicles. We have then developed novel nanovesicle complexes comprising self-assembling mixtures of the nanovesicles, plasmid DNA or siRNA, and targeting peptide ligands. Their biophysical properties were studied and their transfection efficiency was investigated. They transfected cells efficiently without any associated cytotoxicity and with targeting specificity, and in vivo they resulted in very high and tumour-specific uptake and in addition, efficiently transfected the lung. The peptide-targeted nanovesicle complexes allow for the specific targeted enhancement of nucleic acid delivery with improved biosafety over liposomal formulations and represent a promising tool to improve our arsenal of safe, non-viral vectors to deliver therapeutic cargos in a variety of disorders.


Occurrence and relative risk of stroke in incident and prevalent contemporary rheumatoid arthritis.
In contrast with the wealth of data on ischaemic heart disease in rheumatoid arthritis (RA), data on stroke are scarce and contradictory. Despite the high clinical and aetiological relevance, there is no data regarding when (if ever) after RA diagnosis there is an increased risk. Our objective was to assess the risk of stroke (by subtype) in contemporary patients with RA, particularly in relation to time since RA diagnosis. One incident RA cohort diagnosed between 1997 and 2009 (n=8077) and one nationwide prevalent RA cohort followed at Swedish rheumatology clinics between 2005 and 2009 ((n=39 065) were assembled). Each cohort member was matched to a general population comparator. Information on first-time hospitalisations for stroke up to 2009 was retrieved from the Swedish Patient Register. HR and 95% CI were estimated using Cox models. In prevalent unselected RA, the HR of ischaemic stroke was 1.29 (95% CI 1.18 to 1.41). In the incident RA cohort, the overall risk increase was small and non-significant (overall HR 1.11, 95% CI 0.95 to 1.30). When stratified by RA disease duration, an increased risk of ischaemic stroke was indeed detectable but only after 10 or more years since RA diagnosis (HR>10 years: 2.33, 95% CI 1.25 to 4.34). Risk of haemorrhagic stroke was increased in prevalent but not in incident RA. The magnitude of stroke risk is lower than for ischaemic heart disease in RA, and the evolvement of this risk from RA diagnosis may be slower. This suggests different driving forces behind these two RA co-morbidities and has implications for the clinical follow-up of patients with RA.


Exercise in type 2 diabetes: to resist or to endure?
There is now evidence that a single bout of endurance (aerobic) or resistance exercise reduces 24 h post-exercise subcutaneous glucose profiles to the same extent in insulin-resistant humans with or without type 2 diabetes. However, it remains to be determined which group would benefit most from specific exercise protocols, particularly with regard to long-term glycaemic control. Acute aerobic exercise first accelerates translocation of myocellular glucose transporters via AMP-activated protein kinase, calcium release and mitogen-activated protein kinase, but also improves insulin-dependent glucose transport/phosphorylation via distal components of insulin signalling (phosphoinositide-dependent kinase 1, TBC1 domain family, members 1 and 4, Rac1, protein kinase C). Post-exercise effects involve peroxisome-proliferator activated receptor-γ coactivator 1α and lead to ATP synthesis, which may be modulated by variants in genes such as NDUFB6. While mechanisms of acute resistance-type exercise are less clear, chronic resistance training activates the mammalian target of rapamycin/serine kinase 6 pathway, ultimately increasing protein synthesis and muscle mass. Over the long term, adherence to rather than differences in metabolic variables between specific modes of regular exercise might ultimately determine their efficacy. Taken together, studies are now needed to address the variability of individual responses to long-term resistance and endurance training in real life.