Nudging tourists to donate for conservation: Experimental evidence on soliciting voluntary contributions for coastal management.
This research examines the donation behavior of tourists who are asked to donate to coastal conservation aimed at addressing a bundled mix of land and sea issues. Historically, the governance and financing of land and sea conservation have been separated; yet coastal tourism directly involves a mix of activities and development challenges which link land and sea together on the coast. Marine parks, and numerous studies examining their funding schemes, have typically focused on mandatory user fees targeting specific types of activities. For example, many studies focus only on scuba divers' willingness to pay (WTP) for marine conservation. Alternative funding mechanisms, such as voluntary contributions, may be preferred, or even necessary, to traditional government imposed fees, but much less is known about effective implementation. Relatively few studies focus on bundled cross-boundary conservation activities (i.e., land and sea conservation) from all visitors of a marine park (i.e., beachgoers, surfers, boaters, snorkelers) and its encompassing coastal area. In this study we target tourists visiting a popular island and employ field experimental methods to explore the optimal donation request mechanism and pricing levels that influence real voluntary payments for conservation. The field experiment examines voluntary payments under the treatment conditions: open-ended, a set of several suggested donation amounts, and default opt-in and opt-out at two price levels. The field experiment was conducted with tourists on the island of Gili Trawangan, Indonesia. Results reveal that tourists are willing to donate to bundled land-sea conservation issues and that there is a significantly higher propensity to donate in all treatment conditions compared to the open-ended condition. The default opt-out conditions garnered the highest rate of donations at 75% and 62% respectively for the lower and higher set amounts. The mean donation amount was largest in the higher default opt-out condition. Our results suggest that the optimal method of requesting voluntary donations is a set default amount requiring users to opt-out if they do not wish to donate. Implementing a default opt-out eco-donation targeting all types of visitors represents a significant source of funding and illustrates the potential for donations to finance land and sea conservation efforts, an important avenue for future investigation in many interconnected systems that have been historically governed and financed separately.


Antidepressant effect in older depressed patients: the lessons of two agomelatine trials.
The present paper reports in parallel the findings of the two phase III trials that evaluated the efficacy of agomelatine in older depressed patients. It describes how the particular methodological innovations (particularly in relation to patient selection, design and accuracy of diagnosis of depression) introduced in study 2 have improved the quality of recruitment of patients and the assay sensitivity. Study 1 lacked assay sensitivity, and among the many differences with study 2, the inclusion of unexpected mildly ill patients could have inflated the placebo response. The increased demands on investigators in study 2 appear to have reduced the placebo effect and showed a robust benefit of agomelatine. The two agomelatine studies offer the opportunity to discuss hypotheses that have been raised to explain the low level of response of older patients to available antidepressants.


"Role magnets"? An empirical investigation of popularity trajectories for life-course persistent individuals during adolescence.
Recent scholarship has focused on the role of social status in peer groups to explain the fact that delinquency is disproportionately committed during adolescence. Yet, the precise mechanism linking adolescence, social status, and antisocial behavior is not well understood. Dual-taxonomy postulates a testable mechanism that links the sudden increase in risky behavior among adolescents to the social magnetism of a small group of persistently antisocial individuals, referred to here as the "role magnet" hypothesis. Using semi-parametric group-based trajectory modeling and growth-curve modeling, this study provides the first test of this hypothesis by examining physical violence and popularity trajectories for 1,845 male respondents age 11-32 from a nationally representative sample (54 % non-Hispanic White; 21 % non-Hispanic African American; 17 % Hispanic; 8 % Asian). Individuals assigned to a "chronic violence" trajectory group showed consistently lower average levels of popularity from 11 to 19. But, these same individuals experienced increases in popularity during early adolescence and subsequent declines in late adolescence. These findings are linked to current research examining social status as a mechanism generating antisocial behavior during adolescence and the consequences of delayed entry into adult roles.


Reconstitution of Functionalized Transmembrane Domains of Receptor Proteins into Biomimetic Membranes.
For integral membrane proteins, an assessment of their structures and interactions within a biomimetic lipid bilayer environment is critical for evaluating their cellular function. Hydrophobic sequences prevalent within transmembrane domains, however, make these proteins susceptible to aggregation and, thus, create difficulties in examining their structural and functional properties via canonical techniques. Working exclusively with single-pass transmembrane (TM) segments of bitopic membrane proteins, in the form of soluble peptides, bypasses many of the pitfalls of full-length protein preparations while allowing for the opportunity to examine the properties of TM domains within biomimetic membrane environments. In this study, peptides mimicking the TM domains of the epidermal growth factor receptor (EGFR) and CD4 co-receptor, both cell-signaling surface receptors, have been reconstituted into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) lipid bilayers. The formation of their native α-helical structures within vesicle membranes was observed from circular dichroism, and full partition of the peptides into the membrane was demonstrated by tryptophan fluorescence and neutron reflectivity (NR). Using an engineered planar lipid bilayer system ideal for surface characterization methods, such as surface plasmon resonance (SPR) and NR, the TM peptides, functionalized with a N-terminal biotin tag, proved capable of "activating" a membrane surface, as evidenced by the capture of streptavidin. On the basis of these initial assessments, we anticipate these membrane-bound peptides will provide a versatile platform for understanding the intricate roles of receptor TM domains in cell signaling.


Splicing modulators elicit global translational repression by condensate-prone proteins translated from introns.
Chemical splicing modulators that bind to the spliceosome have provided an attractive avenue for cancer treatment. Splicing modulators induce accumulation and subsequent translation of a subset of intron-retained mRNAs. However, the biological effect of proteins containing translated intron sequences remains unclear. Here, we identify a number of truncated proteins generated upon treatment with the splicing modulator spliceostatin A (SSA) via genome-wide ribosome profiling and bio-orthogonal noncanonical amino acid tagging (BONCAT) mass spectrometry. A subset of these truncated proteins has intrinsically disordered regions, forms insoluble cellular condensates, and triggers the proteotoxic stress response through c-Jun N-terminal kinase (JNK) phosphorylation, thereby inhibiting the mTORC1 pathway. In turn, this reduces global translation. These findings indicate that creating an overburden of condensate-prone proteins derived from introns represses translation and prevents further production of harmful truncated proteins. This mechanism appears to contribute to the antiproliferative and proapoptotic activity of splicing modulators.


How Long Non-Coding RNAs and MicroRNAs Mediate the Endogenous RNA Network of Head and Neck Squamous Cell Carcinoma: a Comprehensive Analysis.
Long non-coding RNAs (lncRNAs) act as competing endogenous RNAs (ceRNAs) to compete for microRNAs (miRNAs) in cancer metastasis. Head and neck squamous cell carcinoma (HNSCC) is one of the most common human cancers and rare biomarkers could predict the clinical prognosis of this disease and its therapeutic effect. Weighted gene co-expression network analysis (WGCNA) was performed to identify differentially expressed mRNAs (DEmRNAs) that might be key genes. GO enrichment and protein-protein interaction (PPI) analyses were performed to identify the principal functions of the DEmRNAs. An lncRNA-miRNA-mRNA network was constructed to understand the regulatory mechanisms in HNSCC. The prognostic signatures of mRNAs, miRNAs, and lncRNAs were determined by Gene Expression Profiling Interactive Analysis (GEPIA) and using Kaplan-Meier survival curves for patients with lung squamous cell carcinoma. We identified 2,023 DEmRNAs, 1,048 differentially expressed lncRNAs (DElncRNAs), and 82 differentially expressed miRNAs (DEmiRNAs). We found that eight DEmRNAs, 53 DElncRNAs, and 16 DEmiRNAs interacted in the ceRNA network. Three ceRNAs (HCG22, LINC00460 and STC2) were significantly correlated with survival. STC2 transcript levels were significantly higher in tumour tissues than in normal tissues, and the STC2 expression was slightly upregulated at different stages of HNSCC. LINC00460, HCG22 and STC2 exhibited aberrant levels of expression and may participate in the pathogenesis of HNSCC.


The bicolored white-toothed shrew Crocidura leucodon (HERMANN 1780) is an indigenous host of mammalian Borna disease virus.
Borna disease (BD) is a sporadic neurologic disease of horses and sheep caused by mammalian Borna disease virus (BDV). Its unique epidemiological features include: limited occurrence in certain endemic regions of central Europe, yearly varying disease peaks, and a seasonal pattern with higher disease frequencies in spring and a disease nadir in autumn. It is most probably not directly transmitted between horses and sheep. All these features led to the assumption that an indigenous virus reservoir of BDV other than horses and sheep may exist. The search for such a reservoir had been unsuccessful until a few years ago five BDV-infected shrews were found in a BD-endemic area in Switzerland. So far, these data lacked further confirmation. We therefore initiated a study in shrews in endemic areas of Germany. Within five years 107 shrews of five different species were collected. BDV infections were identified in 14 individuals of the species bicolored white-toothed shrew (Crocidura leucodon, HERMANN 1780), all originating from BD-endemic territories. Immunohistological analysis showed widespread distribution of BDV antigen both in the nervous system and in epithelial and mesenchymal tissues without pathological alterations. Large amounts of virus, demonstrated by presence of viral antigen in epithelial cells of the oral cavity and in keratinocytes of the skin, may be a source of infection for natural and spill-over hosts. Genetic analyses reflected a close relationship of the BDV sequences obtained from the shrews with the regional BDV cluster. At one location a high percentage of BDV-positive shrews was identified in four consecutive years, which points towards a self-sustaining infection cycle in bicolored white-toothed shrews. Analyses of behavioral and population features of this shrew species revealed that the bicolored white-toothed shrew may indeed play an important role as an indigenous host of BDV.


The heat shock protein 90 inhibitor, (-)-epigallocatechin gallate, has anticancer activity in a novel human prostate cancer progression model.
(-)-Epigallocatechin gallate (EGCG), a major tea polyphenol, elicits anticancer effects. However, the mechanism of action is not fully understood. Our laboratory previously showed that EGCG inhibits heat shock protein 90 (HSP90). We used nontumorigenic (NT), tumorigenic, and metastatic cancer cells from a novel human prostate cancer progression model to test the hypotheses that certain stages are more or less sensitive to EGCG and that sensitivity is related to HSP90 inhibition. Treatment of cells with EGCG, novobiocin, or 17-AAG resulted in more potent cytotoxic effects on tumorigenic and metastatic cells than NT cells. When tumorigenic or metastatic cells were grown in vivo, mice supplemented with 0.06% EGCG in drinking water developed significantly smaller tumors than untreated mice. Furthermore, EGCG prevented malignant transformation in vivo using the full prostate cancer model. To elucidate the mechanism of EGCG action, we performed binding assays with EGCG-Sepharose, a C-terminal HSP90 antibody, and HSP90 mutants. These experiments revealed that EGCG-Sepharose bound more HSP90 from metastatic cells compared with NT cells and binding occurred through the HSP90 C-terminus. In addition, EGCG bound HSP90 mutants that mimic both complexed and uncomplexed HSP90. Consistent with HSP90 inhibitory activity, EGCG, novobiocin, and 17-AAG induced changes in HSP90-client proteins in NT cells and larger differences in metastatic cells. These data suggest that EGCG may be efficacious for the treatment of prostate cancer because it preferentially targets cancer cells and inhibits a molecular chaperone supportive of the malignant phenotype.


Loricaria cuffyi (Siluriformes: Loricariidae), a new species of loricariin catfish from the Guiana Shield.
Loricaria cuffyi n. sp. is described based on 36 specimens from the Essequibo and upper Negro River drainages in western Guyana and the upper Orinoco River drainage in Venezuela. The new species can be distinguished from sympatric and geographically proximate congeners by a postorbital notch that is inconspicuous, shallow and rounded, odontode ridges on the dorsum of head and predorsal weakly developed, abdominal plates tightly joined and completely covering the median abdominal space and pectoral girdle, higher anterior lateral plate counts, and coloration characteristics. The distribution of the new species adds to an interesting and well-documented biogeographical pattern exhibited by other Guiana Shield loricariids influenced by the proto-Berbice during the Cenozoic and recent configuration of drainages in the Guiana Shield. We present an update on the taxonomy of Loricaria, and discuss the biogeography and conservation status of the new species.


Establishing the potency of N-acyl amino acids versus conventional fatty acids as thermogenic uncouplers in cells and mitochondria from different tissues.
The possibility that N-acyl amino acids could function as brown or brite/beige adipose tissue-derived lipokines that could induce UCP1-independent thermogenesis by uncoupling mitochondrial respiration in several peripheral tissues is of significant physiological interest. To quantify the potency of N-acyl amino acids versus conventional fatty acids as thermogenic inducers, we have examined the affinity and efficacy of two pairs of such compounds: oleate versus N-oleoyl-leucine and arachidonate versus N-arachidonoyl-glycine in cells and mitochondria from different tissues. We found that in cultures of the muscle-derived L6 cell line, as well as in primary cultures of murine white, brite/beige and brown adipocytes, the N-acyl amino acids were proficient uncouplers but that they did not systematically display higher affinity or potency than the conventional fatty acids, and they were not as efficient uncouplers as classical protonophores (FCCP). Higher concentrations of the N-acyl amino acids (as well as of conventional fatty acids) were associated with signs of deleterious effects on the cells. In liver mitochondria, we found that the N-acyl amino acids uncoupled similarly to conventional fatty acids, thus apparently via activation of the adenine nucleotide transporter-2. In brown adipose tissue mitochondria, the N-acyl amino acids were able to activate UCP1, again similarly to conventional fatty acids. We thus conclude that the formation of the acyl-amino acid derivatives does not confer upon the corresponding fatty acids an enhanced ability to induce thermogenesis in peripheral tissues, and it is therefore unlikely that the N-acyl amino acids are of specific physiological relevance as UCP1-independent thermogenic compounds.


A comparison of two emergency medical dispatch protocols with respect to accuracy.
Emergency medical dispatching should be as accurate as possible in order to ensure patient safety and optimize the use of ambulance resources. This study aimed to compare the accuracy, measured as priority level, between two Swedish dispatch protocols - the three-graded priority protocol Medical Index and a newly developed prototype, the four-graded priority protocol, RETTS-A. A simulation study was carried out at the Emergency Medical Communication Centre (EMCC) in Stockholm, Sweden, between October and March 2016. Fifty-three voluntary telecommunicators working at SOS Alarm were recruited nationally. Each telecommunicator handled 26 emergency medical calls, simulated by experienced standard patients. Manuscripts for the scenarios were based on recorded real-life calls, representing the six most common complaints. A cross-over design with 13 + 13 calls was used. Priority level and medical condition for each scenario was set through expert consensus and used as gold standard in the study. A total of 1293 calls were included in the analysis. For priority level, n = 349 (54.0%) of the calls were assessed correctly with Medical Index and n = 309 (48.0%) with RETTS-A (p = 0.012). Sensitivity for the highest priority level was 82.6% (95% confidence interval: 76.6-87.3%) in the Medical Index and 54.0% (44.3-63.4%) in RETTS-A. Overtriage was 37.9% (34.2-41.7%) in the Medical Index and 28.6% (25.2-32.2%) in RETTS-A. The corresponding proportion of undertriage was 6.3% (4.7-8.5%) and 23.4% (20.3-26.9%) respectively. In this simulation study we demonstrate that Medical Index had a higher accuracy for priority level and less undertriage than the new prototype RETTS-A. The overall accuracy of both protocols is to be considered as low. Overtriage challenges resource utilization while undertriage threatens patient safety. The results suggest that in order to improve patient safety both protocols need revisions in order to guarantee safe emergency medical dispatching.


Insulin/IGF-like signalling, the central nervous system and aging.
Enormous strides in understanding aging have come from the discovery that mutations in single genes can extend healthy life-span in laboratory model organisms such as the yeast Saccharomyces, the fruit fly Drosophila melanogaster, the nematode worm Caenorhabditis elegans and the mouse. IIS [insulin/IGF (insulin-like growth factor)-like signalling] stands out as an important, evolutionarily conserved pathway involved in the determination of lifespan. The pathway has diverse functions in multicellular organisms, and mutations in IIS can affect growth, development, metabolic homoeostasis, fecundity and stress resistance, as well as lifespan. The pleiotropic nature of the pathway and the often negative effects of its disruption mean that the extent, tissue and timing of IIS manipulations are determinants of a positive effect on lifespan. One tissue of particular importance for lifespan extension in diverse organisms is the CNS (central nervous system). Although lowered IIS in the CNS can extend lifespan, IIS is also widely recognized as being neuroprotective and important for growth and survival of neurons. In the present review, we discuss our current understanding of the role of the nervous system in extension of lifespan by altered IIS, and the role of IIS in determination of neuronal function during aging. The nervous system can play both endocrine and cell-autonomous roles in extension of lifespan by IIS, and the effects of IIS on lifespan and neuronal function can be uncoupled to some extent. Tissue-specific manipulation of IIS and the cellular defence mechanisms that it regulates will better define the ways in which IIS affects neuronal and whole-organism function during aging.


Introduction of Human Flt3-L and GM-CSF into Humanized Mice Enhances the Reconstitution and Maturation of Myeloid Dendritic Cells and the Development of Foxp3+CD4+ T Cells.
Two cytokines, fms-related tyrosine kinase 3 ligand (Flt3-L) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are considered to be the essential regulators of dendritic cell (DC) development in vivo. However, the combined effect of Flt3-L and GM-CSF on human DCs has not been evaluated in vivo. In this study, we, therefore, aimed at evaluating this using a humanized mouse model. Humanized non-obese diabetic/SCID/Jak3null (hNOJ) mice were constructed by transplanting hematopoietic stem cells from human umbilical cord blood into newborn NOJ mice, and in vivo transfection (IVT) was performed by hydrodynamic injection-mediated gene delivery using plasmids encoding human Flt3-L and GM-CSF. Following IVT, Flt3-L and GM-CSF were successfully induced in hNOJ mice. At 10 days post-IVT, we found, in the spleen, that treatment with both Flt3-L and GM-CSF enhanced the reconstitution of two myeloid DC subsets, CD14-CD1c+ conventional DCs (cDCs) and CD14-CD141+ cDCs, in addition to CD14+ monocyte-like cells expressing CD1c and/or CD141. GM-CSF alone had less effect on the reconstitution of these myeloid cell populations. By contrast, none of the cytokine treatments enhanced CD123+ plasmacytoid DC (pDC) reconstitution. Regardless of the reconstitution levels, three cell populations (CD1c+ myeloid cells, CD141+ myeloid cells, and pDCs) could be matured by treatment with cytokines, in terms of upregulation of CD40, CD80, CD86, and CD184/CXCR4 and downregulation of CD195/CCR5. In particular, GM-CSF contributed to upregulation of CD80 in all these cell populations. Interestingly, we further observed that Foxp3+ cells within splenic CD4+ T cells were significantly increased in the presence of GM-CSF. Foxp3+ T cells could be subdivided into two subpopulations, CD45RA-Foxp3hi and CD45RA-Foxp3lo T cells. Whereas CD45RA-Foxp3hi T cells were increased only after treatment with GM-CSF alone, CD45RA-Foxp3lo T cells were increased only after treatment with both Flt3-L and GM-CSF. Treatment with Flt3-L alone had no effect on the number of Foxp3+ T cells. The correlation analysis demonstrated that the development of these Foxp3+ subpopulations was associated with the maturation status of DC(-like) cells. Taken together, this study provides a platform for studying the in vivo effect of Flt3-L and GM-CSF on human DCs and regulatory T cells.


A Longitudinal Investigation of Children's Trauma Memory Characteristics and Their Relationship with Posttraumatic Stress Disorder Symptoms.
While trauma memory characteristics are considered a core predictor of adult PTSD, the literature on child PTSD is limited and inconsistent. We investigated whether children's trauma memory characteristics predict their posttraumatic stress symptoms (PTSS) at 1 month and 6 months post-trauma. We recruited 126 6-13 year olds who experienced a single-incident trauma that led to attendance at an emergency department. We assessed trauma memory disorganisation and sensory-emotional qualities through both narrative recall and self-report questionnaire, and PTSS at 1-month post-trauma and at 6-month follow-up. We found that, after controlling for age, children's self-reported trauma memory characteristics were positively associated with their concurrent PTSS, and longitudinally predicted symptoms 6-months later. However, observable trauma memory characteristics coded from children's narratives were not related to PTSS at any time. This suggests that children's perceptions of their trauma memories are a more reliable predictor of the development and maintenance of PTSS than the nature of their trauma narrative, which has important implications for clinical practice.


Diarrhea after kidney transplantation: a new look at a frequent symptom.
Diarrhea is a frequent but overlooked complication of kidney transplantation. Diarrhea is repeatedly neglected, often considered by patients and clinicians an unavoidable side effect of immunosuppressive regimens. It is, however, associated with a significant impairment in life quality. Severe and chronic posttransplant diarrhea may lead to dehydration, malabsorption, rehospitalization, immunosuppression, noncompliance, and a greater risk of graft loss and death. There is thus a need to optimize and standardize the management of posttransplant diarrhea with consistent diagnostic and therapeutic strategies. A recent study has suggested that the increased sensitivity of molecular tools might help in early pathogen identification and guidance of antimicrobial treatment. Most bacterial and protozoan infections are readily curable with appropriate antimicrobial agents; cryptosporidiosis and C. difficile infections may however be complicated by relapsing courses. In addition, identification of enteric viral genomes in stool has further reduced posttransplant diarrhea of unknown origin. Chronic norovirus-related posttransplant diarrhea, arising from the interplay of the virus and immunosuppressive drugs, has emerged as a new challenge in the field. Prospective and controlled studies are necessary to evaluate the efficacy and safety of innovative anti-norovirus therapeutics, as well as optimal immunosuppressive regimens, to enable viral clearance while preventing rejection and donor-specific antibody formation. This review seeks to provide a basis for the design of future clinical prospective studies.


Iatrogenic genitourinary fistula: an 18-year retrospective review of 805 injuries.
Genitourinary fistula poses a public health challenge in areas where women have inadequate access to quality emergency obstetric care. Fistulas typically develop during prolonged, obstructed labor, but providers can also inadvertently cause a fistula when performing obstetric or gynecological surgery. This retrospective study analyzes 805 iatrogenic fistulas from a series of 5,959 women undergoing genitourinary fistula repair in 11 countries between 1994 and 2012. Injuries fall into three categories: ureteric, vault, and vesico-[utero]/-cervico-vaginal. This analysis considers the frequency and characteristics of each type of fistula and the risk factors associated with iatrogenic fistula development. In this large series, 13.2 % of genitourinary fistula repairs were for injuries caused by provider error. A range of cadres conducted procedures resulting in iatrogenic fistula. Four out of five iatrogenic fistulas developed following surgery for obstetric complications: cesarean section, ruptured uterus repair, or hysterectomy for ruptured uterus. Others developed during gynecological procedures, most commonly hysterectomy. Vesico-[utero]/-cervico-vaginal fistulas were the most common (43.6 %), followed by ureteric injuries (33.9 %) and vault fistulas (22.5 %). One quarter of women with iatrogenic fistulas had previously undergone a laparotomy, nearly always a cesarean section. Among these women, one quarter had undergone more than one previous cesarean section. Women with previous cesarean sections are at an increased risk of iatrogenic injury. Work environments must be adequate to reduce surgical error. Training must emphasize the importance of optimal surgical techniques, obstetric decision-making, and alternative ways to deliver dead babies. Iatrogenic fistulas should be recognized as a distinct genitourinary fistula category.


The olfactory responses of the antenna and maxillary palp of the fleshfly, Neobellieria bullata (Diptera: Sarcophagidae), and their sensitivity to blockage of nitric oxide synthase.
The relative sensitivities of the olfactory receptors in the antenna and maxillary palp of the fleshfly, Neobellieria bullata, were assessed using simultaneous electroantennograms (EAGs) and electropalpograms (EPGs). In general, the antennae and maxillary palps were more sensitive to odors related to animals (blood extract and saturated carboxylic acid) than to odors that were plant-derived (citral, hexenol, hexenal). In addition, the maxillary palps were relatively less sensitive to plant-derived odorants than the antennae, perhaps related to their anatomical position. Scanning electron microscopy was also used to assess the types of sensilla found on the two organs. In addition, NADPH-diaphorase histochemistry was used in an attempt to localize the enzyme nitric oxide synthase (NOS) in the antenna and the maxillary palps. We found evidence of NADPH-diaphorase staining in both organs, with localized staining in the antennal cells and more general staining in the maxillary palps. When NOS was selectively blocked using the antagonist L-NAME, the amplitude of the EAGs and EPGs to odorants fell by 30-50%. In contrast, application of the inactive enantiomer, D-NAME, did not change the amplitude of the EAGs or the EPGs. Our results indicate that NOS is involved in the function of olfactory receptor cells in the fleshfly.


Optogenetic modulation of TDP-43 oligomerization accelerates ALS-related pathologies in the spinal motor neurons.
Cytoplasmic aggregation of TDP-43 characterizes degenerating neurons in most cases of amyotrophic lateral sclerosis (ALS). Here, we develop an optogenetic TDP-43 variant (opTDP-43), whose multimerization status can be modulated in vivo through external light illumination. Using the translucent zebrafish neuromuscular system, we demonstrate that short-term light stimulation reversibly induces cytoplasmic opTDP-43 mislocalization, but not aggregation, in the spinal motor neuron, leading to an axon outgrowth defect associated with myofiber denervation. In contrast, opTDP-43 forms pathological aggregates in the cytoplasm after longer-term illumination and seeds non-optogenetic TDP-43 aggregation. Furthermore, we find that an ALS-linked mutation in the intrinsically disordered region (IDR) exacerbates the light-dependent opTDP-43 toxicity on locomotor behavior. Together, our results propose that IDR-mediated TDP-43 oligomerization triggers both acute and long-term pathologies of motor neurons, which may be relevant to the pathogenesis and progression of ALS.


Absence of age effects on spontaneous past and future thinking in daily life.
Previous research on voluntary mental time travel (i.e., deliberately thinking about the past or future) has resulted in negative age effects. In contrast, studies on spontaneous past thoughts (i.e., involuntary autobiographical memories) have reported small or no age effects. The aim of the present research was to investigate the effects of age on the nature and frequency of spontaneous future thoughts in everyday life. In two studies, we examined whether older adults reported spontaneous future thoughts as often as younger adults, and whether these thoughts were predominantly goal-oriented and less dependent on incidental cues than thoughts about the past. In Study 1, young and old participants kept a diary of spontaneous thoughts of upcoming prospective memory tasks and involuntary autobiographical memories for 2 weeks. In Study 2, a 1-day experience sampling method was used to investigate spontaneous and deliberate task-unrelated future and past thoughts, by having young and old participants complete a questionnaire in response to 30 random signals. In both studies, no age effects were found in the frequency of spontaneous future thoughts, which were reported as frequently as thoughts about the past (Study 2) and referred predominantly to upcoming tasks and goals rather than future events or hypothetical events (Study 2). Results concerning the triggers of reported thoughts provide initial support for the idea that representations of future thoughts may be more activated than memories of past events. Taken together, these results have important implications for research on cognitive ageing, mind-wandering, prospective memory and involuntary autobiographical memories.


Activation of the Brain to Postpone Dementia: A Concept Originating from Postmortem Human Brain Studies.
Alzheimer's disease (AD) is characterized by decreased neuronal activity and atrophy, while hyperactivity of neurons seems to make them resistant to aging and neurodegeneration, a phenomenon which we have paraphrased as 'use it or lose it'. Our hypothesis proposes that (1) during their functioning, neurons are damaged; (2) accumulation of damage that is not repaired is the basis of aging; (3) the vulnerability to AD is determined by the genetic background and the balance between the amount of damage and the efficiency of repair, and (4) by stimulating the brain, repair mechanisms are stimulated and cognitive reserve is increased, resulting in a decreased rate of aging and risk for AD. Environmental stimulating factors such as bilingualism/multilingualism, education, occupation, musical experience, physical exercise, and leisure activities have been reported to reduce the risk of dementia and decrease the rate of cognitive decline, although methodological problems are present.


Polyamine-based analogs and conjugates as antikinetoplastid agents.
Naturally occurring polyamines: putrescine, spermidine and spermine are crucial for Kinetoplastid growth and persistence. These aliphatic polycations are either biosynthesized or internalized into Kinetoplastid by active transport. Impairing the polyamine metabolism using polyamine derivatives is an interesting path in the search of new antikinetoplastid chemotherapy. In the past 30 years, research interest in this field has been constantly expanding and recent results demonstrated that the discovery of a polyamine-based antikinetoplastid drug is undoubtedly possible. In this paper, all the polyamine derivatives previously described to present an antikinetoplastid activity are reported. This review is organized around three main parts which are diamine, triamine and tetramine derivatives. Each part includes the description of the series of molecules and, their in vitro and in vivo activity when available. Structure-activity relationships of these derivatives are discussed and the most promising structures for a positive outcome are eventually highlighted.


Maternal Psychosocial Maladjustment and Child Internalizing Symptoms: Investigating the Modulating Role of Maternal Sensitivity.
In light of evidence suggesting that maternal adaptation may impact early child emotional development, this study investigated the interactive effects of maternal psychosocial maladjustment and maternal sensitivity on child internalizing symptoms, with the aim of investigating the potentially protective function of maternal sensitivity. Families (N = 71 to 106 across measures, with gender spread almost evenly: number of boys = 31 to 51 across measures) took part in four assessments between child ages 1 and 3 years. Mothers completed measures of parental stress, psychological distress, and marital satisfaction when their children were between 12 and 15 months. A composite score of maternal psychosocial maladjustment was derived from these measures. Maternal sensitivity was rated by trained observers at 12 months following a home visit. Child internalizing symptoms were assessed by both parents when the child was 2 and 3 years old. Hierarchical regressions revealed that increased maternal psychosocial maladjustment was related to more internalizing symptoms in children, however only among children of less sensitive mothers. In contrast, children of more sensitive mothers appeared to be protected. This was observed with maternal reports at 2 years, and both maternal and paternal reports at 3 years. These results suggest that young children may be differentially affected by their parents' emotional adjustment, while highlighting the pivotal protective role of maternal sensitivity in this process.


Soil microbiome responses to the short-term effects of Amazonian deforestation.
Slash-and-burn clearing of forest typically results in increase in soil nutrient availability. However, the impact of these nutrients on the soil microbiome is not known. Using next generation sequencing of 16S rRNA gene and shotgun metagenomic DNA, we compared the structure and the potential functions of bacterial community in forest soils to deforested soils in the Amazon region and related the differences to soil chemical factors. Deforestation decreased soil organic matter content and factors linked to soil acidity and raised soil pH, base saturation and exchangeable bases. Concomitant to expected changes in soil chemical factors, we observed an increase in the alpha diversity of the bacterial microbiota and relative abundances of putative copiotrophic bacteria such as Actinomycetales and a decrease in the relative abundances of bacterial taxa such as Chlamydiae, Planctomycetes and Verrucomicrobia in the deforested soils. We did not observe an increase in genes related to microbial nutrient metabolism in deforested soils. However, we did observe changes in community functions such as increases in DNA repair, protein processing, modification, degradation and folding functions, and these functions might reflect adaptation to changes in soil characteristics due to forest clear-cutting and burning. In addition, there were changes in the composition of the bacterial groups associated with metabolism-related functions. Co-occurrence microbial network analysis identified distinct phylogenetic patterns for forest and deforested soils and suggested relationships between Planctomycetes and aluminium content, and Actinobacteria and nitrogen sources in Amazon soils. The results support taxonomic and functional adaptations in the soil bacterial community following deforestation. We hypothesize that these microbial adaptations may serve as a buffer to drastic changes in soil fertility after slash-and-burning deforestation in the Amazon region.


Effect of aspirin and other NSAIDs on postmenopausal breast cancer incidence by hormone receptor status: results from a prospective cohort study.
Aspirin and other non-aspirin non-steroidal anti-inflammatory drugs (NSAIDs) can inhibit aromatase activity and thus could selectively lower incidence of hormone receptor positive tumors. We assessed whether the association of aspirin and other NSAIDs with postmenopausal breast cancer risk differs by estrogen and progesterone receptor (ER and PR) status of the tumor. A population-based cohort of 26,580 postmenopausal women was linked to a SEER Cancer Registry to identify incident breast cancers. Regular use of aspirin and other NSAIDs was reported on a self-administered questionnaire mailed in 1992. Cox proportional hazards models were used to estimate multivariate relative risks (RRs) and 95% confidence intervals (CIs) of breast cancer incidence overall and by ER and PR status, adjusting for multiple breast cancer risk factors. Through 2005, 1,581 incident breast cancer cases were observed. Compared to aspirin never users, women who regularly consumed aspirin had a lower risk of breast cancer (RR = 0.80; 95% CI: 0.71-0.90), and there was evidence for lower risk with increasing frequency of use (RR = 0.71 for aspirin use 6 or more times/week vs. never use; P trend = 0.00001). Inverse associations for regular aspirin use were observed for ER+ (RR = 0.77; 95% CI 0.67-0.89), ER- (RR = 0.78; 95% CI 0.56-1.08), PR+ (RR = 0.79; 95% CI 0.68-0.92), and PR- (RR = 0.73; 95% CI 0.56-0.95) breast cancers. In contrast, use of other NSAIDs was not associated with breast cancer incidence overall (RR = 0.95, 95% CI: 0.85-1.07), or by ER or PR status. Aspirin, but not other NSAID use, was associated with about 20% lower risk of postmenopausal breast cancer and did not vary by ER or PR status of the tumor, suggesting that the hypothesized protective effects of aspirin may either be through cellular pathways independent of estrogen or progesterone signaling, or on tumor microenvironment.


Increased leptin levels correlate with thyroid autoantibodies in nonobese males.
Leptin is an adipokine that regulates body weight and appetite. It is also an inflammatory cytokine that influences immune reactivity and autoimmunity. Leptin levels are increased in obesity and are higher in women than in men. We aimed to determine whether leptin levels, independent of sex and body mass index (BMI), are associated with thyroid autoimmunity. This study uses data from The Third National Health and Nutrition Examination Survey (NHANES III) to test the association of leptin and thyroid autoimmunity, independent of BMI. Thyroid-stimulating hormone, thyroxine, antithyroid peroxidase (TPO) antibodies and leptin levels were measured in 2902 men and 3280 women within the NHANES III population. BMI was calculated from height and weight. Women had significantly higher leptin levels and anti-TPO antibody titres than men. Correlation analyses demonstrated that leptin levels were associated with anti-TPO antibody levels in the total population, but when men and women were analysed separately, this association was lost. We then stratified men and women into obese (BMI > 30) or nonobese (BMI ≤ 30) subgroups and determined the association between leptin levels and anti-TPO antibody titres for each subgroup. Using regression analysis, we found that increased leptin levels correlated with thyroid autoantibodies in nonobese males, but not in obese males or in females. Leptin levels correlated with thyroid autoantibody titres in nonobese males. This association was not found in females. Sex and body habitus should therefore be considered in studying the role of leptin in other autoimmune conditions.


African American Patient Preferences for Glaucoma Education.
The objectives of the study were to examine (a) the types of questions that African American patients have about glaucoma for their providers and (b) how patients' sociodemographic characteristics are associated with where and from whom they would like to learn about glaucoma and glaucoma medications. Forty-nine adult African American patients with glaucoma were recruited at a private ophthalmology clinic where they completed a questionnaire for this cross-sectional study. African American patients had a mean of 3.9 questions for their ophthalmologists; the questions that patients checked as having most often were "What is my prognosis with glaucoma?" (49%) and "What is my intraocular pressure?" (45%). Seventy-six percent of patients preferred that an educational program about glaucoma be offered at the doctor's office and 39% preferred it be offered at a community or senior citizen center. Ninety percent said that the education program should be offered by doctors. Patients under the age of 70 were significantly more likely to want a program on the Internet than patients age 70 and over (Pearson χ = 4.7, P = .03). If an educational program was developed patients reported being most interested in the following topics, glaucoma medications (84%), what is glaucoma and what does it mean to have it? (83%). African American patients have many questions about glaucoma for their eye care providers. African American patients would prefer glaucoma educational programs be offered at their provider's office. Our findings could be used to develop educational programs for African American patients with glaucoma.


Opportunistic bilateral salpingectomy during benign gynecological surgery for ovarian cancer prevention: a survey of Gynecologic Oncology Committee of Japan Society of Obstetrics and Gynecology.
Recent evidence has supported the concept that epithelial ovarian cancer (EOC) arises from the cells of the fallopian tube or endometrium. This study investigated current practice in Japan with respect to performing opportunistic bilateral salpingectomy (OBS) during gynecological surgery for benign disease for Ovarian Cancer Prevention. We mailed a questionnaire to 767 hospitals and clinics, comprising 628 accredited training institutions of the Japan Society of Obstetrics and Gynecology (JSOG), Japan Society of Gynecologic Oncology (JSGO), or Japan Society of Gynecologic and Obstetric Endoscopy and Minimally Invasive Therapy (JSGOE) and 139 private institutions with at least one JSGOE-certified licensed gynecologic laparoscopist. Among the 767 institutions, 444 (57.9%) provided responses, including 91 (20.6%) that were both JSGOE and JSGO accredited, 71 (16.0%) that were only JSGO accredited, 88 (19.8%) that were only JSGOE accredited, and 194 (43.7%) that were unaccredited. It was found that awareness and performance of OBS largely depended on the JSGO and/or JSGOE accreditation status. OBS was only performed at 54.0% of responding institutions and just 6.8% of the institutions were willing to participate in randomized controlled trials to validate this method for reducing the incidence of ovarian cancer. The JSOG Gynecologic Tumor Committee will announce its opinion on salpingectomy for ovarian cancer prevention to all JSOG members and will develop a system for monitoring the number of OBS procedures in Japan.


Effects of electroacupuncture on stress and anxiety-related responses in rats.
The aim of this work was to investigate if eletroacupuncture at PC6 would modulate the stress-induced anxiety-like behavior and the level of activation of several brain areas. Rats were distributed in groups: control; submitted to immobilization; submitted to immobilization and eletroacupuncture at PC6 or at the tail. Immobilization increased grooming and decreased stretched attend postures and the time spent in the open arms of the ele-vated plus-maze. Eletroacupuncture at PC6 or tail canceled the effect of immobilization on grooming and attenuated the stretched attend posture. Immobilization increased Fos-immunoreactivity in the prefrontal cortex, medial and central amygdala, paraventricular and dorsomedial nuclei of the hypothalamus, lateral hypothalamus, dentate gyrus, CA1, CA2 and CA3 hippocampal areas. The activation of paraventricular, dorsomedial nuclei and prefrontal cortex by immobilization was canceled by electroacupuncture at PC6 and attenuated by electroacupuncture in the tail. The activation of the other areas was canceled by electroacupuncture in PC6 or the tail. It is concluded that immobilization induced anxiety-like behavior that was moderately attenuated by eletroacupuncture with difference between the stimulation in PC6 or the rat tail. Eletroacupuncture showed specificity concerning to the attenuation of the effects of immobilization in the CNS areas related to the stress response, anxiety and cardiovascular system.


[Efficacy of Arnica in varicose vein surgery: results of a randomized, double-blind, placebo-controlled pilot study].
In homeopathy ARNICA is widely used as a woundhealing medication and for the treatment of hematomas. In this pilot study the efficacy and safety of ARNICA D12 in patients following varicose vein surgery were investigated. Prospective, randomized, double-blind, placebo-controlled pilot trial according to ICH GCP guidelines. The study was conducted by a surgeon at the Angiosurgical Clinic, Berlin- Buch. After randomized allocation, 60 patients received either ARNICA D12 or placebo. Start of medication occurred the evening before operation with 5 globules. On the operation day one preoperative and hourly postoperative dosages after awakening were given. On days 2-14 of the study 5 globules 3 times a day were given. OUTCOME CRITERIA: Surface (in cm(2) and using a three-point verbal rating scale) and intensity of hematomas induced by operation, complications of wound healing, and intensity of pain (five-point verbal rating scale) as well as efficacy and safety of the study medication were assessed. Hematoma surface was reduced (from day 7 to day 14) under ARNICA by 75.5% and under placebo by 71.5% (p = 0.4726). The comparison of hematoma surface (small, medium, large) using the verbal rating scale yielded a value of p = 0.1260. Pain score decreased by 1.0 +/- 2.2 points under ARNICA and 0.3 +/- 0.8 points under placebo (p = 0.1977). Remission or improvement of pain was observed in 43.3% of patients in the ARNICA group and in 27.6% of patients in the placebo group. Tolerability was rated as very good in all cases. The results of this pilot study showed a trend towards a beneficial effect of ARNICA D12 with regard to reduction of hematoma and pain during the postoperative course. For a statistically significant proof of efficacy of ARNICA D12 in patients following varicose vein surgery a larger sample size is necessary.


Polarization and statistical analysis of scenes containing a semireflector.
We present an approach to recover scenes deteriorated by reflections off a semireflecting medium (e.g., a glass window). The method, based on imaging through a polarizer at two or more orientations, separates the reflected and transmitted scenes and determines which is which. We analyze the polarization effects, taking into account internal reflections within the medium. The scene reconstruction requires the estimation of the orientation (inclination and tilt angles) of the transparent (invisible) surface. The inclination angle is estimated by seeking the value that leads to the minimal mutual information of the estimated scenes. The limitations and the consequences of noise and angle error are discussed, including a fundamental ambiguity in the determination of the plane of incidence. Experimental results demonstrate the success of angle estimation and consequent scene separation and labeling.


Cancer incidences in Europe related to mortalities, and ethnohistoric, genetic, and geographic distances.
We have previously shown that geographic differences in cancer mortalities in Europe are related to (in order of importance): geographic distances (reflecting environmental differences), ethnohistoric distances (encompassing cultural and genetic attributes), and genetic distances of the populations in the areas studied. In this study, we analyzed the relations of the same three factors to European incidences of 45 male and 47 female cancers. Differences in cancer incidences are correlated moderately, first with geographic distances, and then with genetic distances, but not at all with ethnohistoric distances. Comparing these findings to the earlier ones for cancer mortalities, we note the reversal in the importance of ethnohistory and genetics, and the generally lower correlations of incidence differences with the three putatively causal distance matrices. A path diagram combining both studies demonstrates the lack of cultural carcinogenic effects, but suggests cultural influences on procedures such as the registration of deaths in different political entities. Additionally, the relatively large correlation between ethnohistoric distances and mortality differences is caused by common factors behind the correlation of ethnohistoric and geographic distances. Geographic proximity results in similar ethnohistories. The direct effects of genetic distances are negligible and only their common effects with geographic distances play a role, accounting for the weak to negligible influence of genetics on incidence and mortality differences. Apparently, the genetic systems available to us do not substantially affect cancer incidence or mortality. We present indirect evidence that international differences in the quality of cancer rate data are greater in mortalities than in incidences.


The acute effects of exposure to the electromagnetic field emitted by mobile phones on human attention.
The aim of our study was to investigate the effects of acute mobile phone exposure on a range of tasks which tapped capacity and processing speed within the attentional system. Thirty-eight healthy volunteers were randomly assigned to either an experimental group which was exposed to a connected mobile phone or a control group in which the mobile phone was switched off. Subjects remained blind to mobile phone status throughout duration of study. The experimental group were exposed to an electromagnetic field emitted by a 900 MHz mobile phone for 30 min. Cognitive performance was assessed at three points (prior to mobile phone exposure, at 15 and 30 min post-exposure) using six cognitive neuropsychological tests (digit span and spatial span forwards and backwards, serial subtraction and verbal fluency). Significant differences between the two groups were evident after 5 min on two tests of attentional capacity (digit span forwards and spatial span backwards) and one of processing speed (serial subtraction). In all three instances, performance was facilitated following mobile phone exposure. No deficits were evident. These findings are discussed in terms of possible functional and neuroanatomical bases.


Energy-efficient optical line terminal for WDM-OFDM-PON based on two-dimensional subcarrier and layer allocation.
We propose and experimentally demonstrate a scheme to reduce the energy consumption of optical line terminal (OLT) in wavelength division multiplexing - orthogonal frequency division multiplexing - passive optical networks (WDM-OFDM-PONs). In our scheme, a wireless communication technique, termed layered modulation, is introduced to maximize the transmission capacity of OFDM modulation module in the OLT by multiplexing data from different ONU groups with signal-to-noise ratio (SNR) margins onto the same subcarriers. With adaptive and dynamic subcarrier and layer allocation, several ONU groups with low traffic demands can share one OFDM modulation module to deliver their data during non-peak hours of a day, thus greatly reducing the number of running devices and minimizing the energy consumption of the OLT. Numerical calculation shows that an energy efficiency improvement of 28.3% in the OLT can be achieved by using proposed scheme compared to the conventional WDM-OFDM-PON.


Glucocorticoid-mediated control of the activation and clonal deletion of peripheral T cells in vivo.
Poly- and oligoclonal T cell stimuli like anti-CD3 epsilon monoclonal antibody or Staphylococcus aureus enterotoxin B (SEB), injected at doses that per se are not lethal, provoke acute death within less than 24 h, provided that endogenous glucocorticoids (GC) are depleted by adrenalectomy or by injection of saturating amounts of the GC receptor antagonist RU-38486 (mifepristone). Pharmacological doses of the GC agonist dexamethasone (DEX) alter the in vivo response of splenic V beta 8+ T cells to SEB, thus impeding the expansion of such cells and causing their rapid (3 d) clonal deletion. In contrast, coadministration of RU-38486 counteracts a SEB-induced early (12 h) reduction of V beta 8+CD4+ and V beta 8+CD8+ spleen cells. In vivo T cell stimulation by injection of bacterial superantigen induces a rapid (peak at 90-120 min) increase in corticosterone serum levels, suggesting that endogenous GC might control early T cell activation. Accordingly, kinetic studies revealed that RU-38486 has to be administered within 2 h after superantigen administration to exert its lethal effect. Similarly, exogenous GC must be injected during this critical phase (2 h) to rescue animals from acute death induced by coinjection of SEB and D-galactosamine (GalN). Adrenalectomy, injection of RU-38486 and priming with GalN per se provoke the programmed death of peripheral CD4+ and CD8+ T cells. Thus, three manipulations that sensitize mice for the lethal effect of T cell stimulation also exert a proapoptotic effect on peripheral T cells. In synthesis, endogenous and exogenous GC regulate T cell responses and determine the propensity of peripheral T cells to undergo apoptosis.


The prognostic significance of p16(INK4a)/p14(ARF) locus deletion and MDM-2 protein expression in adult acute myelogenous leukemia.
The p16(INK4a) locus encodes two distinct proteins, p16(INK4a) and p14(ARF). Although p16(INK4a) and p15(INK4b) are involved in the phosphorylation of the retinoblastoma (Rb) protein, p14(ARF) interacts with the MDM-2 oncoprotein antagonizing its function as a suppressor of p53. The role of deletions of p16(INK4a)/p14(ARF) and p15(INK4b) and expressions of MDM-2 in myeloid leukemias and its influence on prognosis remain unclear. The authors analyzed deletions of p16(INK4)/p14(ARF) and p15(INK4b) in 74 adults with acute myeloid leukemia (AML) by Southern blotting. Western blotting was used to determine Rb protein phosphorylation in patients with deletions of p16(INK4)/p14(ARF) and p15(INK4b). Then, they analyzed the levels of MDM-2 protein expression and correlated it with prognosis in an expanded population of 79 adults with AML by immunoblot analysis and solid-phase radioimmunoassay. Deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b) occurred in 4 of 74 patients (5%) (hemizygous in 3, homozygous in 1 patient). Although the complete remission (CR) rate was similar (79% vs. 50%; P = 0.187), CR duration (10 vs. 46 weeks; P < 0.001), event free survival rate (EFS; 6 vs. 85 weeks; P < 0.004) and overall survival rate (11 vs. 86 weeks; P = 0.001) were significantly shorter in patients with deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b). Thirty-seven (47%) of 79 patients studied for MDM-2 showed increased MDM-2 expression. These patients had a significantly shorter EFS rate (50 vs. 64 weeks; P = 0.023) and a trend for shorter CR duration (24 vs. 53 weeks; P = 0.07). Overall survival rate was not significantly different (50 vs. 84 weeks; P = 0.136). The authors concluded that 1) deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b) occur with low incidence in patients with AML; 2) patients with deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b) have a significantly shorter CR duration, EFS rate, and overall survival rate than do patients without deletions; (3) overexpression of MDM-2 is common in AML and is associated with shorter CR duration and EFS rate. Mechanisms other than p14(ARF) deletion are responsible for MDM-2 overexpression, and this overexpression may play a role in the biology of the disease.


A long-term retrospective study on rehabilitation of seabirds in Gran Canaria Island, Spain (2003-2013).
The aims of this study were to analyze the causes of morbidity and mortality in a large population of seabirds admitted to the Tafira Wildlife Rehabilitation Center (TWRC) in Gran Canaria Island, Spain, from 2003 to 2013, and to analyze the outcomes of the rehabilitation process. We included 1,956 seabirds (133 dead on admission and 1,823 admitted alive) in this study. Causes of morbidity were classified into nine categories: light pollution (fallout), fishing gear interaction, crude oil, poisoning/intoxication, other traumas, metabolic/nutritional disorder, orphaned young birds, other causes, and unknown/undetermined. The crude and stratified (by causes of admission) rates of the three final disposition categories (euthanasia Er, unassisted mortality Mr, and release Rr), the time until death, and the length of stay were also studied for the seabirds admitted alive. Yellow-legged Gull (Larus michahellis) was the species most frequently admitted (46.52%), followed by Cory's Shearwater (Calonectris diomedea borealis) (20.09%). The most frequent causes of morbidity were light pollution (fallout) (25.81%), poisoning/intoxication (24.69%), and other traumas (18.14%). The final disposition rates were: Er = 15.35%, Mr = 16.29%, and Rr = 68.34%. The highest Er was observed in the 'other traumas' category (58.08%). Seabirds admitted due to metabolic/nutritional disorder had the highest Mr (50%). The highest Rr was observed in the light pollution (fallout) category (99.20%). This survey provides useful information for the conservation of several seabird species. We suggest that at least the stratified analysis by causes of admission of the three final disposition rates, and the parameters time until death and length of stay at the center should be included in the outcome research of the rehabilitation of seabirds. The high release rate for seabirds (68.34%) achieved at the TWRC emphasizes the importance of wildlife rehabilitation centers for the conservation of seabirds.


Proteomic changes in the cortex membrane fraction of genetic absence epilepsy rats from Strasbourg.
Epilepsy is a serious neurodegenerative disorder with a high incidence and a variety of presentations and causes. Studies on brain from various animal models including chronic models: Genetic Absence Epilepsy Rats from Strasbourg (GAERS) are very useful for understanding the fundamental mechanisms associated with human epilepsy. Individual regions of the brain have different protein composition in different conditions. Therefore, proteomic analyses of the brain compartments are preferred for the development of new therapeutic targets in different pathophysiological conditions like neurodegenerative disorders. In this study, we describe a proteomic profiling of membrane fraction of cortex tissue from epileptic GAERS and non-epileptic Wistar rat brain by two-dimensional gel electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectroscopy. Comparing the optical density of spots between groups, we found that one protein expression was significantly down-regulated (guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1) and one protein expression was significantly up-regulated (14-3-3 protein epsilon isoform) in GAERS group. Our results indicate that these proteins might have played a significant role in epilepsy and may be considered as valuable therapeutic targets in the absence of epilepsy.


Direct binding of TRAF2 and TRAF6 to TICAM-1/TRIF adaptor participates in activation of the Toll-like receptor 3/4 pathway.
Using yeast two-hybrid screening, we found three TRAF proteins TRAF1, 2 and 6, bound the N-terminal region of the TLR3/4 adaptor TICAM-1 (TRIF). TRAF2, a newly identified TICAM-1-binding protein, bound the PxQxS motif (aa 333-338) of TICAM-1 using mutagenesis by alanine substitutions. TICAM-1 is known to induce the activation of NF-kappaB and IRF-3, which leads to activation of the interferon (IFN)-beta promoter, an activity that is conserved in the N+TIR fragment (aa 1-533). By mutation of the two distinct binding sites for TRAF2 and TRAF6 in N+TIR TICAM-1, the induction of IFN-beta was completely abrogated. Although the TRAF2 site single mutation only marginally affected TICAM-1-mediated type I IFN induction, it further impaired the function of the TRAF6 site mutant. Moreover, double point mutations of the TRAF2 and TRAF6 binding motifs in TICAM-1 N+TIR reduced the activation of IRF-3 and NF-kappaB, the critical transcription factors for IFN-beta expression. Furthermore, TRAF2/6 functioned as an E3 ligase to induce K63-mediated ubiquitination on N+TIR which was abrogated in the mutant lacking the TRAF2/6 sites in parallel with IFN-inducing activity. Confocal microscopy analysis indicated that TRAF2 and TRAF6 merged with oligomerized (i.e. activated) TICAM-1 N+TIR. However, TRAF3, which is another TRAF family member essential for TLR3-mediated type-I IFN signaling, still assembled in the mutant lacking the TRAF2/6 sites. Our data suggest that the binding of TRAF2 and TRAF6 to TICAM-1 cooperatively activates the IFN-inducing pathway through ubiquitination of TICAM-1, a modification which occurs unrelated to TRAF3 recruitment in the TICAM-1 signaling complex. TRAF2/6 may participate in TICAM-1-mediated IFN-beta induction besides TRAF3.


Cambodian Phellinus linteus inhibits experimental metastasis of melanoma cells in mice via regulation of urokinase type plasminogen activator.
Phellinus linteus (PL) is a fungus mainly found in tropical America, Africa and Asian countries including Korea, Japan and China. PL has been traditionally used for the treatment of arthritis, liver damage and cancer. However, little was known on the biological activity and characterization of Phellinus species in Cambodia. Thus, in the present study, the anti-metastatic mechanism of aqueous extract of Cambodian Phellinus linteus (CPL) was evaluated. Cambodian mushroom was identified as a Phellinus species with 99% homology of Phellinus linteus by DNA sequence analysis and comparison by the National Center for Biotechnology Information. CPL did not exhibit any significant cytotoxicity against B16BL6 cells, invasive melanoma cells at 1 mg/ml. However, CPL inhibited platelet aggregation induced by B16BL6 cells and also disrupted the adhesion to gelatin and invasion of B16BL6 cells in a concentration dependent manner. Similarly, CPL dose-dependently inhibited the pulmonary metastatic colonies in C57BL/6 mice intravenously injected by B16BL6 cells up to 55.5% at a dose of 50 mg/kg compared with untreated control. CPL also down-regulated the expression of urokinase type plasminogen activator (uPA), one of key proteins associated with invasion and metastasis of tumor cells in a concentration dependent fashion, while CPL didn't significantly affect the expression of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 2 (TIMP-2) by reverse transcriptase-polymerase chain reaction (RT-PCR). Taken together, these findings indicate that Cambodian Phellinus linteus may inhibit metastasis at least partly via regulation of uPA associated with tumor cell induced platelet aggregation (TCIPA) and also suggest a further study for isolation of active ingredients and the involvement of adhesion molecule signaling pathway.


High TCL1 expression and intact T-cell receptor signaling define a hyperproliferative subset of T-cell prolymphocytic leukemia.
The T-cell leukemia 1 (TCL1) oncoprotein is overexpressed by chromosomal rearrangement in the majority of cases of T-cell prolymphocytic leukemia (T-PLL). In vitro, TCL1 can modulate the activity of the serine-threonine kinase AKT, a downstream effector of T-cell receptor (TCR) signaling. In a series of 86 T-PLL tumors, we show that expression of TCR, and levels of TCL1 and activated AKT are adverse prognostic markers. High-level TCL1 in TCR-expressing T-PLL is associated with higher presenting white blood cell counts, faster tumor cell doubling, and enhanced in vitro growth response to TCR engagement. In primary tumors and TCL1-transfected T-cell lines, TCR engagement leads to rapid recruitment of TCL1 and AKT to transient membrane activation complexes that include TCR-associated tyrosine kinases, including LCK. Pharmacologic inhibition of AKT activation alters the localization, stability, and levels of these transient TCL1-AKT complexes and reduces tumor cell growth. Experimental introduction and knockdown of TCL1 influence the kinetics and strength of TCR-mediated AKT activation. We propose that in T-PLL, TCL1 represents a highly regulated, targetable modulator of TCR-mediated AKT growth signaling.


Polarization-independent negative-index metamaterial in the near infrared.
We present and evaluate theoretically and experimentally a design for a negative-index metamaterial that is termed the "Swiss cross" structure. Compared with the established fishnet structure, the proposed design eliminates the drawback of polarization-dependent effective optical parameters. The new design is fabricated by means of e-beam technology and experimentally analyzed using spectroscopic techniques. The thorough comparison with numerical simulations reveals an effective refractive index of n=-1.9 at an operational wavelength of 1400 nm that is independent of the incident polarization. The resonances of the system are comprehensively discussed.


Communication: Feshbach resonances in the water molecule revealed by state-selective spectroscopy.
We employ triple-resonance vibrational overtone excitation to access quasibound states of water from several fully characterized bound states of the molecule. Comparison of the measured dissociation spectra allows a rigorous assignment of rotational quantum numbers J, nuclear spin and parity, and a tentative vibrational characterization of the observed resonances. Their asymmetrical shapes (Fano profiles) reflect interference of dipole moments for transitions to these resonances with that to the dissociative continuum. The assignments and Fano profile parameters of the resonances stand as a benchmark for the extension of accurate quantum-mechanical calculations to activated complexes of water. The narrow widths of some of these resonances indicate that water molecules may survive for as long as up to 60 ps in states above the dissociation threshold. We consider the possible implication of such long-lived states for the kinetics of water dissociation and the OH+H association reaction.


Cytomegalovirus in ulcerative colitis: an evidence-based approach to diagnosis and treatment.
The detection of cytomegalovirus (CMV) in the setting of inflammatory bowel disease often creates confusion whether CMV is a 'bystander' or 'disease.' This review discusses the clinical conundrum of CMV in ulcerative colitis, approach to discriminate infection from disease, and therapeutic considerations (immunosuppressive and anti-CMV treatment). CMV disease should be considered in corticosteroid refractory- dependent and thiopurine refractory disease. Endoscopy may reveal deep punched out ulcers, irregular ulcers, or cobble-stoning. The diagnosis rests on the presence and abundance of viral inclusion bodies on hematoxylin and eosin stain, positive immunohistochemistry, and/or positive tissue polymerase chain reaction. CMV disease is associated with worse outcomes including increased colectomy rates. The timing and duration of antiviral drugs in CMV disease is debatable but depends on the load of CMV in tissue. In high-grade infection, CMV needs to be treated while increasing immunosuppression may work in the setting of low-grade infection. Ganciclovir is the drug of choice for treatment of CMV disease. Tumor necrosis factor inhibitors may be useful for treating underlying disease activity in the setting of CMV. Other emerging therapies include fecal microbiota transplantation. Randomized studies are necessary to define the best timing and duration of anti-CMV therapy.


Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A, and Cel7B from Hypocrea jecorina.
Adsorption of cellulases on the cellulose surface is an integral part of the catalytic mechanism, and a detailed description of the adsorption process is therefore required for a fundamental understanding of this industrially important class of enzymes. However, the mode of adsorption has proven intricate, and several key questions remain open. Perhaps most notably it is not clear whether the adsorbed enzyme is in dynamic equilibrium with the free population or irreversibly associated with no or slow dissociation. To address this, we have systematically investigated adsorption reversibility for two cellobiohydrolases (Cel7A and Cel6A) and one endoglucanase (Cel7B) on four types of pure cellulose substrates. Specifically, we monitored dilution-induced release of adsorbed enzyme in samples that had previously been brought to a steady state (constant concentration of free enzyme). In simple dilution experiments (without centrifugation), the results consistently showed full reversibility. In contrast to this, resuspension of enzyme-substrate pellets separated by centrifugation showed extensive irreversibility. We conclude that these enzymes are in a dynamic equilibrium between free and adsorbed states but suggest that changes in the physical properties of cellulose caused by compaction of the pellet hampers subsequent release of adsorbed enzyme. This latter effect may be pertinent to both previous controversies in the literature on adsorption reversibility and the development of enzyme recycling protocols in the biomass industry.


ALS3 and ALS8 represent a single locus that encodes a Candida albicans adhesin; functional comparisons between Als3p and Als1p.
The ALS (agglutinin-like sequence) gene family of Candida albicans encodes eight cell-surface glycoproteins, some of which are involved in adherence to host surfaces. A mutational analysis of each ALS gene is currently being performed to deduce the functions of the encoded proteins and to better understand the role of these proteins in C. albicans biology and pathogenesis. This paper describes construction of an als3/als3 mutant and comparison of its phenotype to an als1/als1 strain. Efforts to disrupt ALS3 indicated that the gene could be deleted in two transformation steps, suggesting that the gene is encoded by a single locus and that the ALS3-like locus, ALS8, does not exist. Strains lacking ALS3 or ALS1 did not exhibit a defect in germ tube formation when grown in RPMI 1640 medium, but the als1/als1 mutant formed significantly fewer germ tubes in Lee medium. Analysis of ALS3 and ALS1 promoter activity using green fluorescent protein (GFP) reporter strains and flow cytometry showed that when cells are placed into medium that promotes germ tube formation, ALS1 is transcribed prior to ALS3. Comparison of the mutant strains in adhesion assays showed that the als3/als3 strain was defective in adhesion to both human umbilical vein endothelial cells (HUVEC) and buccal epithelial cells (BEC), but not to fibronectin-coated plastic plates. In contrast, the als1/als1 strain showed decreased adherence to HUVEC, but adherence to BEC and fibronectin were the same as wild-type controls. Inoculation of the buccal reconstituted human epithelium (RHE) model of oral candidiasis with the mutant strains showed nearly a total lack of adhesion and epithelial destruction by the als3/als3 mutant while the als1/als1 strain showed only a slightly reduced degree of epithelial destruction compared to the wild-type control. Adhesion data presented here suggest that, in the assays performed, loss of Als3p affects C. albicans adhesion more than loss of Als1p. Collectively, these results demonstrate functional similarities and differences between Als1p and Als3p, and suggest the potential for more complex interrelationships between the ALS genes and their encoded proteins.


Long-range nonlocal flow of vortices in narrow superconducting channels.
We report a new nonlocal effect in vortex matter, where an electric current confined to a small region of a long and sufficiently narrow superconducting wire causes vortex flow at distances hundreds of intervortex separations away. The observed remote traffic of vortices is attributed to a very efficient transfer of a local strain through the one-dimensional vortex lattice (VL), even in the presence of disorder. We also observe mesoscopic fluctuations in the nonlocal vortex flow, which arise due to "traffic jams" when vortex arrangements do not match a local geometry of a superconducting channel.


Effect of individually chosen bed-height adjustments on the low-back stress of nurses.
The effects of height-adjustable beds in hospitals on the subsequent prevalence of low-back problems among nurses depend on the capacity to reduce low-back stress by bed-height adjustment. This capacity was investigated in the present study. Professional nurses performed patient-handling tasks at a standard and an individually chosen bed height. Peak values and time integrals of spinal compression and shear forces were estimated with dynamic biomechanical modeling. The bed-height adjustment led to lower values of time-integrated compression (average 8.8% lower), peak shear force (average 9.3% lower), and time-integrated shear force (average 18.1% lower). No significance was found for the effect on peak compression, nor for the results for each individual task. This finding can be explained by the minor adjustments made in comparison with the standard height or by the application of different criteria for bed-height adjustment. The decreasing time-integrated forces and peak shear force, without a concomitant rise of peak compression, speak in favor of the use of height-adjustable beds in nursing.


The interactome of stabilized α-synuclein oligomers and neuronal proteins.
While it is generally accepted that α-synuclein oligomers (αSOs) play an important role in neurodegeneration in Parkinson's disease, the basis for their cytotoxicity remains unclear. We have previously shown that docosahexaenoic acid (DHA) stabilizes αSOs against dissociation without compromising their ability to colocalize with glutamatergic synapses of primary hippocampal neurons, suggesting that they bind to synaptic proteins. Here, we develop a proteomic screen for putative αSO binding partners in rat primary neurons using DHA-stabilized human αSOs as a bait protein. The protocol involved co-immunoprecipitation in combination with a photoactivatable heterobifunctional sulfo-LC-SDA crosslinker which did not compromise neuronal binding and preserved the interaction between the αSOs-binding partners. We identify in total 29 proteins associated with DHA-αSO of which eleven are membrane proteins, including synaptobrevin-2B (VAMP-2B), the sodium-potassium pump (Na+ /K+ ATPase), the V-type ATPase, the voltage-dependent anion channel and calcium-/calmodulin-dependent protein kinase type II subunit gamma; only these five hits were also found in previous studies which used unmodified αSOs as bait. We also identified Rab-3A as a target with likely disease relevance. Three out of four selected hits were subsequently validated with dot-blot binding assays. In addition, likely binding sites on these ligands were identified by computational analysis, highlighting a diversity of possible interactions between αSOs and target proteins. These results constitute an important step in the search for disease-modifying treatments targeting toxic αSOs.


Bone regeneration and docking site healing after bone transport distraction osteogenesis in the canine mandible.
Bone transport distraction osteogenesis provides a promising alternative to traditional grafting techniques. However, existing bone transport distraction osteogenesis devices have many limitations. The purpose of this research was to test a new device, the mandibular bone transport reconstruction plate, in an animal model with comparable mandible size to humans and to histologically and mechanically examine the regenerate bone. Eleven adult foxhounds were divided into an unreconstructed control group of 5 animals and an experimental group of 6 animals. In each animal, a 34-mm segmental defect was created in the mandible. The defect was reconstructed with a bone transport reconstruction plate. Histologic and biomechanical characteristics of the regenerate and unrepaired defect were analyzed and compared with bone on the contralateral side of the mandible after 4 weeks of consolidation. The reconstructed defect was bridged with new bone, with little bone in the control defect. Regenerate density and microhardness were 22.3% and 42.6%, respectively, lower than the contralateral normal bone. Likewise, the anisotropy of the experimental group was statistically lower than in the contralateral bone. Half the experimental animals showed nonunion at the docking site. The device was very stable and easy to install and activate. After 1 month of consolidation, the defect was bridged with new bone, with evidence of active bone formation. Regenerate bone was less mature than the control bone. Studies are underway to identify when the regenerate properties compare with normal bone and to identify methods to augment bone union at the docking site.


Wide Variability in the Time Required for Immunotoxins to Kill B Lineage Acute Lymphoblastic Leukemia Cells: Implications for Trial Design.
Recombinant immunotoxins (rITs) targeting CD22 are highly active in hairy cell leukemia, but less so in acute lymphoblastic leukemia (ALL). This study aims to understand the variable activity of an rIT against ALL toward improving responses in clinical application. We determined in vitro activity of rITs by WST-8 assays and the time needed to kill ALL cell lines and patient-derived ALL blasts by flow cytometry. The findings were translated into two systemic ALL xenograft models. Differences in time needed to kill KOPN-8 cells for distinct rITs were addressed biochemically. In vitro activity (IC50) of anti-CD22 rIT varied 210-fold from 0.02 to 4.6 ng/mL. Activity also varied greatly depending on the time ALL cells were exposed to immunotoxin from < 30 minutes to > 4 days. For KOPN-8, the difference in exposure time was related to intracellular rIT processing. We showed in newly developed ALL xenograft models, where immunotoxins have a short half-life, that the needed exposure time in vitro predicted the responses in vivo By replacing bolus dose with small doses at frequent intervals or with continuous infusion, responses were substantially improved. We confirmed exposure time variability on patient-derived ALL samples and showed a correlation between exposure time needed to reach maximal cytotoxicity in vitro and their clinical response. The exposure time needed for rITs targeting CD22 to kill ALL cells varies widely. Our results suggest that ALL patients would have a better response rate to anti-CD22 immunotoxins if treated by continuous infusion rather than by bolus injections. Clin Cancer Res; 22(19); 4913-22. ©2016 AACR.


Mek1 Down Regulates Rad51 Activity during Yeast Meiosis by Phosphorylation of Hed1.
During meiosis, programmed double strand breaks (DSBs) are repaired preferentially between homologs to generate crossovers that promote proper chromosome segregation at Meiosis I. In many organisms, there are two strand exchange proteins, Rad51 and the meiosis-specific Dmc1, required for interhomolog (IH) bias. This bias requires the presence, but not the strand exchange activity of Rad51, while Dmc1 is responsible for the bulk of meiotic recombination. How these activities are regulated is less well established. In dmc1Δ mutants, Rad51 is actively inhibited, thereby resulting in prophase arrest due to unrepaired DSBs triggering the meiotic recombination checkpoint. This inhibition is dependent upon the meiosis-specific kinase Mek1 and occurs through two different mechanisms that prevent complex formation with the Rad51 accessory factor Rad54: (i) phosphorylation of Rad54 by Mek1 and (ii) binding of Rad51 by the meiosis-specific protein Hed1. An open question has been why inhibition of Mek1 affects Hed1 repression of Rad51. This work shows that Hed1 is a direct substrate of Mek1. Phosphorylation of Hed1 at threonine 40 helps suppress Rad51 activity in dmc1Δ mutants by promoting Hed1 protein stability. Rad51-mediated recombination occurring in the absence of Hed1 phosphorylation results in a significant increase in non-exchange chromosomes despite wild-type levels of crossovers, confirming previous results indicating a defect in crossover assurance. We propose that Rad51 function in meiosis is regulated in part by the coordinated phosphorylation of Rad54 and Hed1 by Mek1.


A new gene mapping resource: interspecies hybrids between Père David's deer (Elaphurus davidianus) and red deer (Cervus elaphus).
Three male F1 hybrids between Père David's deer and red deer were mated to red deer to produce 143 backcross calves. The pedigrees are a rare example of a fertile hybrid between evolutionarily divergent species. We examined the use of these families for genetic mapping of evolutionarily conserved (Type I) loci by testing for genetic linkage between five species-specific protein variants and 12 conserved DNA probes. Two probes were homologous, and the remainder syntenic, to the protein coding loci in cattle or humans. Using six restriction enzymes, each DNA probe detected one or more restriction fragments specific to Père David's deer. Linkage analyses among the species-specific variants placed the loci into four linkage groups within which linkage between adjacent loci and gene order was supported by a LOD > 3. The linkage groups were (HPX, HBB)-FSHB-ACP2, LDHA-CD5-IGF2, BMP3-(GC, ALB)-(KIT, PDGFRA) and LDLR-C3-FGF1. Southern and protein analysis of LDHA and ALB provided identical segregation data. These linkage groups were consistent with the cattle gene map and provide new information for comparing the gene maps of ruminants, humans and mice. The deer hybrids are an important new resource that can contribute to the comparative analysis of the mammalian genome.


Research trends in human osteology: a content analysis of papers published in the American Journal of Physical Anthropology.
This paper explores recent research trends in human osteology, based on articles published in the American Journal of Physical Anthropology (AJPA) during two 5-year intervals: 1980--1984 and 1996--2000. Topical "visibility" is measured in terms of article counts; "impact" is estimated through citation indices. Our results indicate that human osteologists continue to publish a range of methodological, analytical, and descriptive research papers that address a broad array of subjects. Analytical articles are cited more frequently than descriptive articles and thus have higher impact, reflecting the discipline's continued commitment to problem-oriented research. Differences in publication patterns exist between scholars during early and later stages of their careers. Articles published by students and Ph.D.s within 2 years of their doctoral degree are more frequently descriptive than analytical, when compared to people with longer career histories. Topics such as pathology, forensic anthropology, and biodistance modeling remain highly visible, while articles on the dentition have waned. An increase in functional research directed toward the postcranial skeleton is also reflected in our data. While continued visibility for morphological investigations is apparent, the impact of recently developed applications in bone chemistry and molecular anthropology is amply documented in our data, particularly during the more recent survey years.


Early Exercise in Critically Ill Youth and Children, a Preliminary Evaluation: The wEECYCLE Pilot Trial.
To determine the feasibility of conducting a full trial evaluating the efficacy of early mobilization using in-bed cycling as an adjunct to physiotherapy, on functional outcomes in critically ill children. Single center, pilot, randomized controlled trial. Twelve-bed tertiary care, medical-surgical PICU at McMaster Children's Hospital, Hamilton, ON, Canada. Children 3-17 years old who were limited to bed-rest with an expected PICU stay of at least 48 hours. Patients were excluded if they were at their baseline level of function, already mobilizing out of bed or expected to do so within 24 hours. Patients were randomized in a 2:1 ratio to early mobilization using in-bed cycling in addition to usual care physiotherapy (cycling arm) or to usual care physiotherapy alone (control). Usual care was according to institutional practice guidelines. The primary outcome was feasibility and safety. Thirty patients were enrolled (20 to the cycling and 10 to control) over a 12-month period, at a 93.7% consent rate. The median (interquartile range) time from PICU admission to mobilization was 1.5 days (1-3) in the cycling arm and 2.5 days (2-7) in the control arm. Total duration of mobilization therapy in PICU was 210 (152-380) and 136 minutes (42-314 min) in cycling and control arms, respectively. Total number of PICU days mobilized was 5.0 (3-6) with cycling and 2.5 (2-4.8) with usual care. No adverse events occurred in either arm. The main threat to feasibility of mobilization was the availability of physiotherapists or research personnel. Early mobilization is safe and feasible in the PICU. In-bed cycling may facilitate greater duration and intensity of mobilization, in critically ill children. A full-scale randomized controlled trial is warranted to evaluate the efficacy of this intervention on PICU-acquired morbidities and functional outcomes in this population.


Re: Amelsvoort et al. "Approaches for predicting long-term sickness absence".
We would like to thank Van Amelsvoort et al (1) for the interest in our study (2) and take the opportunity to clarify here that none of the workers were sick-listed when they participated in the baseline health survey. We mentioned in the abstract that incident (ie, not prevalent) long-term sickness absence was retrieved from an occupational health register (2). Our explanation of how to interpret the area under the receiver operating characteristic (ROC) curve as measure of discrimination between workers with and without long-term sickness absence might have given the impression that the study population was a mix of workers with and without sickness absence. Throughout the paper, however, workers with long-term sickness absence refer to those not sick-listed at baseline who had incident long-term sickness absence during 1-year follow-up. We agree with the authors that instruments to predict long-term sickness absence for workers still at work (secondary prevention) should be distinguished from instruments for workers already on sick leave (tertiary prevention). The objective of our study was to investigate the Work Ability Index (WAI) as an instrument to predict future long-term sickness absence in non-sick-listed workers, ie, as an instrument for secondary prevention. Therefore, the term "screening" was used in the appropriate context. Van Amelsvoort et al (1) raise an interesting point when they state that including the outcome (sickness absence) as predictor in the model will shift the focus towards the prediction of recurrent sickness absence. Obviously, sickness absence is useless for predicting the first long-term sickness absence episode of an individual who has just finished education and enters the workforce. During working life, workers develop a sickness absence history either without sickness absence episodes (ie, zero-absenteeism) or with successive sickness absence episodes. In the latter case, Navarro et al (3) recommended to use statistical techniques for recurrent rather than independent events. A worker's sickness absence history is the strongest predictor of future sickness absence episodes (4, 5). From that perspective, it would be a missed opportunity not to include past sickness absence as variable in prediction models for future long-term sickness absence.


Both maximal expression of selenoproteins and selenoprotein deficiency can promote development of type 2 diabetes-like phenotype in mice.
Selenium (Se) is an essential trace element in mammals that has been shown to exert its function through selenoproteins. Whereas optimal levels of Se in the diet have important health benefits, a recent clinical trial has suggested that supplemental intake of Se above the adequate level potentially may raise the risk of type 2 diabetes mellitus. However, the molecular mechanisms for the effect of dietary Se on the development of this disease are not understood. In the present study, we examined the contribution of selenoproteins to increased risk of developing diabetes using animal models. C57BL/6J mice (n=6-7 per group) were fed either Se-deficient Torula yeast-based diet or diets supplemented with 0.1 and 0.4 parts per million Se. Our data show that mice maintained on an Se-supplemented diet develop hyperinsulinemia and have decreased insulin sensitivity. These effects are accompanied by elevated expression of a selective group of selenoproteins. We also observed that reduced synthesis of these selenoproteins caused by overexpression of an i(6)A(-) mutant selenocysteine tRNA promotes glucose intolerance and leads to a diabetes-like phenotype. These findings indicate that both high expression of selenoproteins and selenoprotein deficiency may dysregulate glucose homeostasis and suggest a role for selenoproteins in development of diabetes.


Using optimal control methods with constraints to generate singlet states in NMR.
A method is proposed for optimizing the performance of the APSOC (Adiabatic-Passage Spin Order Conversion) technique, which can be exploited in NMR experiments with singlet spin states. In this technique magnetization-to-singlet conversion (and singlet-to-magnetization conversion) is performed by using adiabatically ramped RF-fields. Optimization utilizes the GRAPE (Gradient Ascent Pulse Engineering) approach, in which for a fixed search area we assume monotonicity to the envelope of the RF-field. Such an approach allows one to achieve much better performance for APSOC; consequently, the efficiency of magnetization-to-singlet conversion is greatly improved as compared to simple model RF-ramps, e.g., linear ramps. We also demonstrate that the optimization method is reasonably robust to possible inaccuracies in determining NMR parameters of the spin system under study and also in setting the RF-field parameters. The present approach can be exploited in other NMR and EPR applications using adiabatic switching of spin Hamiltonians.


A combined protocol for identification of maggots of forensic interest.
In Forensic Entomology the estimation of the age of insects is used for the estimation of the minimum post-mortem interval. As insect development is temperature dependent and species specific, a correct species identification is therefore fundamental. In the majority of cases the molecular identification is based on a destructive approach. In this paper a working protocol for molecular identification of fly larvae without affecting the anatomical characters used for morphological identification is presented. The suggested technique allows the preservation of the larval exoskeleton and of the unused soft tissues in the same vial allowing a repetition of both the morphological and molecular identification and reducing the risk of loss of the evidence. This method also allows the possibility of measuring the size of the specimens before their morphological and biomolecular characterization. In order to demonstrate that this technique can be applied on maggots of a large spectrum of dimensions it has been tested and validated using larvae of different size from ~1.7-1.3cm [Calliphora vomitoria and Lucilia sericata (Diptera: Calliphoridae)] to ~10-6.5mm [Musca domestica (Diptera: Muscidae) and Megaselia scalaris (Diptera: Phoridae)]. The importance of a unique identifier and of a complete database with all the specimen information (origin, sample size, identification, etc.) is also discussed.


Long-acting anticholinergic use in chronic obstructive pulmonary disease: efficacy and safety.
This article reviews findings from recently published randomized controlled trials and systematic reviews to provide an up-to-date assessment of the efficacy and safety of tiotropium, the only currently available long-acting muscarinic antagonist, when used alone or in conjunction with other respiratory medications in the treatment of chronic obstructive pulmonary disease (COPD). Results of recent clinical trials support findings from earlier trials in patients with moderate to very severe COPD demonstrating significant benefits of tiotropium compared to placebo, including sustained increases in lung function, reductions in exacerbations and risk of exacerbation-related hospitalizations, and improvement in health status. These benefits were particularly noted in the 4-year UPLIFT study that included 5993 COPD patients, including a large percentage with moderate severity. Whereas the cardiovascular safety of tiotropium has been questioned, results of the UPLIFT trial and a recent pooled analysis of data from 30 trials of tiotropium demonstrated that tiotropium is associated with reductions in the risk of all-cause mortality, cardiovascular mortality and cardiovascular events. Recent findings confirm the benefits of tiotropium in COPD management and provide reassurance regarding its safety. Moreover, the recent UPLIFT trial provides supportive evidence for the efficacy of tiotropium in COPD patients already receiving treatment with long-acting inhaled beta-agonists and inhaled corticosteroids, suggesting advantages of 'triple' therapy in advanced disease. Further, well designed, adequately powered studies should explore the potential advantages and disadvantages of various combinations of currently available long-acting respiratory medications in COPD, particularly in different clinical phenotypes of this heterogeneous disease.


Cloning and reexpression of a functional human IgM anti-lipid A antibody.
The human hybridoma cell line HR78 secretes a human antibody of the IgM isotype directed against bacterial endotoxin. The cell line produces low levels of antibody and, more importantly, the antibody product is likely to be impure since two and perhaps more species of heavy chain are being synthesized as judged by cloning and expression studies. To address these issues, the cell line was used as a source of mRNA for the construction of cDNA libraries and the subsequent isolation of the sequences encoding heavy and light chains. Expression of these chains in a non-immunoglobulin producing murine hybridoma cell line resulted in a monoclonal antibody that bound antigen in a manner essentially identical to that observed for the parental antibody. Moreover, the level of expression of human IgM in selected clones was increased approximately 50-fold over the parental cell line and was comparable to or better than antibody expression by a typical murine hybridoma.


Adolescent peer relations, friendships, and romantic relationships: do they predict social anxiety and depression?
This study examined multiple levels of adolescents' interpersonal functioning, including general peer relations (peer crowd affiliations, peer victimization), and qualities of best friendships and romantic relationships as predictors of symptoms of depression and social anxiety. An ethnically diverse sample of 421 adolescents (57% girls; 14 to 19 years) completed measures of peer crowd affiliation, peer victimization, and qualities of best friendships and romantic relationships. Peer crowd affiliations (high and low status), positive qualities in best friendships, and the presence of a dating relationship protected adolescents against feelings of social anxiety, whereas relational victimization and negative interactions in best friendships predicted high social anxiety. In contrast, affiliation with a high-status peer crowd afforded some protection against depressive affect; however, relational victimization and negative qualities of best friendships and romantic relationships predicted depressive symptoms. Some moderating effects for ethnicity were observed. Findings indicate that multiple aspects of adolescents' social relations uniquely contribute to feelings of internal distress. Implications for research and preventive interventions are discussed.