Mediator-dependent recruitment of TFIIH modules in preinitiation complex.
In vitro, without Mediator, the association of general transcription factors (GTF) and RNA polymerase II (Pol II) in preinitiation complexes (PIC) occurs in an orderly fashion. In this work, we explore the in vivo function of Mediator in GTF recruitment to PIC. A direct interaction between Med11 Mediator head subunit and Rad3 TFIIH subunit was identified. We explored the significance of this interaction and those of Med11 with head module subunits Med17 and Med22 and found that impairing these interactions could differentially affect the recruitment of TFIIH, TFIIE, and Pol II in the PIC. A med11 mutation that altered promoter occupancy by the TFIIK kinase module of TFIIH genome-wide also reduced Pol II CTD serine 5 phosphorylation. We conclude that the Mediator head module plays a critical role in TFIIH and TFIIE recruitment to the PIC. We identify steps in PIC formation that suggest a branched assembly pathway.


Antidepressants and the risk of suicidal behaviors.
The relation between use of antidepressants, especially selective serotonin reuptake inhibitors (SSRIs), and suicidal ideation and behaviors has received considerable public attention recently. The use of such drugs among teenagers has been of particular concern. To estimate the relative risks (RRs) of nonfatal suicidal behavior in patients starting treatment with 1 of 3 antidepressant drugs compared with patients starting treatment with dothiepin. Matched case-control study of patients treated in UK general practices using the UK General Practice Research Database for 1993-1999. The base population included 159,810 users of the 4 antidepressant drugs. Participants could have used only 1 of these antidepressants and had to have received at least 1 prescription for the study antidepressant within 90 days before their index date (the date of suicidal behavior or ideation for cases and the same date for matched controls). Frequency of first-time exposure to amitriptyline, fluoxetine, paroxetine, and dothiepin of patients with a recorded diagnosis of first-time nonfatal suicidal behavior or suicide compared with comparable patients who did not exhibit suicidal behavior. After controlling for age, sex, calendar time, and time from first antidepressant prescription to the onset of suicidal behavior, the relative risks for newly diagnosed nonfatal suicidal behavior in 555 cases and 2062 controls were 0.83 (95% confidence interval, [CI] 0.61-1.13) for amitriptyline, 1.16 (95% CI, 0.90-1.50) for fluoxetine, and 1.29 (95% CI, 0.97-1.70) for paroxetine compared with those using dothiepin. The RR for suicidal behavior among patients first prescribed an antidepressant within 1 to 9 days before their index date was 4.07 (95% CI, 2.89-5.74) compared with patients who were first prescribed an antidepressant 90 days or more before their index date. Time since first antidepressant prescription was not, however, a confounder of the relation between specific antidepressants and suicidal behavior since its relation to suicidal behavior was not materially different among users of the 4 study drugs. Similarly for fatal suicide, the RR among patients who were first prescribed an antidepressant within 1 to 9 days before their index date was 38.0 (95% CI, 6.2-231) compared with those who were first prescribed an antidepressant 90 days or more before their index date. There were no significant associations between the use of a particular study antidepressant and the risk of suicide. The risk of suicidal behavior after starting antidepressant treatment is similar among users of amitriptyline, fluoxetine, and paroxetine compared with the risk among users of dothiepin. The risk of suicidal behavior is increased in the first month after starting antidepressants, especially during the first 1 to 9 days. A possible small increase in risk (bordering statistical significance) among those starting the newest antidepressant, paroxetine, is of a magnitude that could readily be due to uncontrolled confounding by severity of depression. Based on limited information, we also conclude that there is no substantial difference in effect of the 4 drugs on people aged 10 to 19 years.


A non-paternalistic value theory of occupational therapy: a comment to Dige.
In an article by Morten Dige in the Scandinavian Journal of Occupational Therapy, a value theory for occupational therapy is developed where the goals of occupational therapy, i.e. activity and participation, are seen as valuable in themselves. Such a value theory opens up for a paternalistic approach towards the person in need of occupational therapy who risks being excluded from certain types of activities and the theory even risks excluding certain groups of persons. In this comment to Dige, it is argued that adopting a more subjective approach to value, in which activity and participation are instrumental to values such as subjective well-being or the fulfilment of desires, the risk of inherent paternalism and exclusion is minimized. Moreover, activity and participation can still maintain their central role as goals of occupational therapy.


Nutrigenomics and nutrigenetics: the emerging faces of nutrition.
The recognition that nutrients have the ability to interact and modulate molecular mechanisms underlying an organism's physiological functions has prompted a revolution in the field of nutrition. Performing population-scaled epidemiological studies in the absence of genetic knowledge may result in erroneous scientific conclusions and misinformed nutritional recommendations. To circumvent such issues and more comprehensively probe the relationship between genes and diet, the field of nutrition has begun to capitalize on both the technologies and supporting analytical software brought forth in the post-genomic era. The creation of nutrigenomics and nutrigenetics, two fields with distinct approaches to elucidate the interaction between diet and genes but with a common ultimate goal to optimize health through the personalization of diet, provide powerful approaches to unravel the complex relationship between nutritional molecules, genetic polymorphisms, and the biological system as a whole. Reluctance to embrace these new fields exists primarily due to the fear that producing overwhelming quantities of biological data within the confines of a single study will submerge the original query; however, the current review aims to position nutrigenomics and nutrigenetics as the emerging faces of nutrition that, when considered with more classical approaches, will provide the necessary stepping stones to achieve the ambitious goal of optimizing an individual's health via nutritional intervention.


Enhanced activation of cellular AMPK by dual-small molecule treatment: AICAR and A769662.
AMP-activated protein kinase (AMPK) is a key cellular energy sensor and regulator of metabolic homeostasis. Activation of AMPK provides beneficial outcomes in fighting against metabolic disorders such as insulin resistance and type 2 diabetes. Currently, there is no allosteric AMPK activator available for the treatment of metabolic diseases, and limited compounds are available to robustly stimulate cellular/tissue AMPK in a specific manner. Here we investigated whether simultaneous administration of two different pharmacological AMPK activators, which bind and act on different sites, would result in an additive or synergistic effect on AMPK and its downstream signaling and physiological events in intact cells. We observed that cotreating primary hepatocytes with the AMP mimetic 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) and a low dose (1 μM) of the allosteric activator A769662 produced a synergistic effect on AMPK Thr172 phosphorylation and catalytic activity, which was associated with a more profound increase/decrease in phosphorylation of downstream AMPK targets and inhibition of hepatic lipogenesis compared with single-compound treatment. Mechanistically, we found that cotreatment does not stimulate LKB1, upstream kinase for AMPK, but it protects against dephosphorylation of Thr172 phosphorylation by protein phosphatase PP2Cα in an additive manner in a cell-free assay. Collectively, we demonstrate that AICAR sensitizes the effect of A769662 and promotes AMPK activity and its downstream events. The study demonstrates the feasibility of promoting AMPK activity by using two activators with distinct modes of action in order to achieve a greater activation of AMPK and downstream signaling.


Dissemination of spotted fever rickettsia agents in Europe by migrating birds.
Migratory birds are known to play a role as long-distance vectors for many microorganisms. To investigate whether this is true of rickettsial agents as well, we characterized tick infestation and gathered ticks from 13,260 migratory passerine birds in Sweden. A total of 1127 Ixodes spp. ticks were removed from these birds and the extracted DNA from 957 of them was available for analyses. The DNA was assayed for detection of Rickettsia spp. using real-time PCR, followed by DNA sequencing for species identification. Rickettsia spp. organisms were detected in 108 (11.3%) of the ticks. Rickettsia helvetica, a spotted fever rickettsia associated with human infections, was predominant among the PCR-positive samples. In 9 (0.8%) of the ticks, the partial sequences of 17kDa and ompB genes showed the greatest similarity to Rickettsia monacensis, an etiologic agent of Mediterranean spotted fever-like illness, previously described in southern Europe as well as to the Rickettsia sp.IrITA3 strain. For 15 (1.4%) of the ticks, the 17kDa, ompB, gltA and ompA genes showed the greatest similarity to Rickettsia sp. strain Davousti, Rickettsia japonica and Rickettsia heilongjiangensis, all closely phylogenetically related, the former previously found in Amblyomma tholloni ticks in Africa and previously not detected in Ixodes spp. ticks. The infestation prevalence of ticks infected with rickettsial organisms was four times higher among ground foraging birds than among other bird species, but the two groups were equally competent in transmitting Rickettsia species. The birds did not seem to serve as reservoir hosts for Rickettsia spp., but in one case it seems likely that the bird was rickettsiemic and that the ticks had acquired the bacteria from the blood of the bird. In conclusion, migratory passerine birds host epidemiologically important vector ticks and Rickettsia species and contribute to the geographic distribution of spotted fever rickettsial agents and their diseases.


Midlife milk consumption and substantia nigra neuron density at death.
To examine the relationship between midlife milk intake and Parkinson disease (PD) incidence through associations with substantia nigra (SN) neuron density and organochlorine pesticide exposure in decedent brains from the Honolulu-Asia Aging Study. Milk intake data were collected from 1965 to 1968 in 449 men aged 45-68 years with postmortem examinations from 1992 to 2004. Neuron density (count/mm(2)) was measured in quadrants from a transverse section of the SN. Additional measures included brain residues of heptachlor epoxide, an organochlorine pesticide found at excessively high levels in the milk supply in Hawaii in the early 1980s. Neuron density was lowest in nonsmoking decedents who consumed high amounts of milk (>16 oz/d). After removing cases of PD and dementia with Lewy bodies, adjusted neuron density in all but the dorsomedial quadrant was 41.5% lower for milk intake >16 oz/d vs intake that was less (95% confidence interval 22.7%-55.7%, p < 0.001). Among those who drank the most milk, residues of heptachlor epoxide were found in 9 of 10 brains as compared to 63.4% (26/41) for those who consumed no milk (p = 0.017). For those who were ever smokers, an association between milk intake and neuron density was absent. Milk intake is associated with SN neuron loss in decedent brains unaffected by PD. Whether contamination of milk with organochlorine pesticides has a role in SN neurodegeneration warrants further study.


Loss of the glycine N-methyltransferase gene leads to steatosis and hepatocellular carcinoma in mice.
Glycine N-methyltransferase (GNMT) is the main enzyme responsible for catabolism of excess hepatic S-adenosylmethionine (SAMe). GNMT is absent in hepatocellular carcinoma (HCC), messenger RNA (mRNA) levels are significantly lower in livers of patients at risk of developing HCC, and GNMT has been proposed to be a tumor-susceptibility gene for liver cancer. The identification of several children with liver disease as having mutations of the GNMT gene further suggests that this enzyme plays an important role in liver function. In the current study we studied development of liver pathologies including HCC in GNMT-knockout (GNMT-KO) mice. GNMT-KO mice have elevated serum aminotransferase, methionine, and SAMe levels and develop liver steatosis, fibrosis, and HCC. We found that activation of the Ras and Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathways was increased in liver tumors from GNMT-KO mice coincidently with the suppression of the Ras inhibitors Ras-association domain family/tumor suppressor (RASSF) 1 and 4 and the JAK/STAT inhibitors suppressor of cytokine signaling (SOCS) 1-3 and cytokine-inducible SH2-protein. Finally, we found that methylation of RASSF1 and SOCS2 promoters and the binding of trimethylated lysine 27 in histone 3 to these 2 genes was increased in HCC from GNMT-KO mice. These data demonstrate that loss of GNMT induces aberrant methylation of DNA and histones, resulting in epigenetic modulation of critical carcinogenic pathways in mice.


Lack of seasonal and moult-related stress modulation in an opportunistically breeding bird: The white-plumed honeyeater (Lichenostomus penicillatus).
This article is part of a Special Issue "SBN 2014". In most vertebrate species, glucocorticoid levels and stress sensitivity vary in relation to season and life-history stage. In birds, baseline corticosterone (CORT) and stress sensitivity are typically highest while breeding and decrease substantially during moult. Because elevated CORT adversely affects protein synthesis, moult-related CORT suppression is thought to be necessary for forming high-quality feathers. Surprisingly, some passerine species lack moult-related CORT suppression, but these are distinguished by having slow rates of moult and being opportunistic breeders. We examined baseline and stress-induced CORT levels in an opportunistically breeding Australian passerine, the white-plumed honeyeater (Lichenostomus penicillatus). Although this species has a slower moult rate than high-latitude breeders, it differs little from north-temperate passerines. Neither baseline nor stress-induced CORT levels varied with season (winter, spring or summer), sex or moult status in adult birds. While breeding tended to be highest in early spring through late summer, laparotomies revealed only limited reduction in testicular size in males the year round. In all but one sampling period, at least some females displayed follicular hierarchy. Breeding usually coincides with outbreaks of phytophagous insects, which can happen at any time of the year. This results in moult/breeding overlap when infestations occur in late spring or summer. The ability of this species to moult and breed at the same time while having breeding-levels of CORT demonstrates that CORT suppression is not a prerequisite for synthesis of high-quality feathers. An experimental design incorporating moulting and non-moulting phenotypes is suggested to test the functional significance of CORT suppression in other species.


Exact score distribution computation for ontological similarity searches.
Semantic similarity searches in ontologies are an important component of many bioinformatic algorithms, e.g., finding functionally related proteins with the Gene Ontology or phenotypically similar diseases with the Human Phenotype Ontology (HPO). We have recently shown that the performance of semantic similarity searches can be improved by ranking results according to the probability of obtaining a given score at random rather than by the scores themselves. However, to date, there are no algorithms for computing the exact distribution of semantic similarity scores, which is necessary for computing the exact P-value of a given score. In this paper we consider the exact computation of score distributions for similarity searches in ontologies, and introduce a simple null hypothesis which can be used to compute a P-value for the statistical significance of similarity scores. We concentrate on measures based on Resnik's definition of ontological similarity. A new algorithm is proposed that collapses subgraphs of the ontology graph and thereby allows fast score distribution computation. The new algorithm is several orders of magnitude faster than the naive approach, as we demonstrate by computing score distributions for similarity searches in the HPO. It is shown that exact P-value calculation improves clinical diagnosis using the HPO compared to approaches based on sampling. The new algorithm enables for the first time exact P-value calculation via exact score distribution computation for ontology similarity searches. The approach is applicable to any ontology for which the annotation-propagation rule holds and can improve any bioinformatic method that makes only use of the raw similarity scores. The algorithm was implemented in Java, supports any ontology in OBO format, and is available for non-commercial and academic usage under: https://compbio.charite.de/svn/hpo/trunk/src/tools/significance/


Practical management of imatinib in gastrointestinal stromal tumors.
Gastrointestinal stromal tumors (GISTs) have an incidence of 7-15 occurrences per million people. Tyrosine kinase inhibitors (TKIs) have significantly improved clinical outcomes as part of multidisciplinary disease management. The authors will review developments in the management of GISTs, including diagnosis, risk stratification, prognosis, and treatment with imatinib. Imatinib is recommended for postsurgical adjuvant therapy and, where appropriate, neoadjuvant therapy. Clinical practice guidelines recommend first-line imatinib for metastatic and unresectable GISTs based on trials showing efficacy at the standard dose (400 mg per day) and at higher doses of 600-800 mg per day. Oncology nurses play a key role in patient management through (a) patient education about GISTs and their treatment including the use of imatinib, (b) timely scheduling of radiologic follow-up to assess treatment response, (c) monitoring treatment adherence, (d) helping to sustain imatinib dose intensity by monitoring toxicities and drug interactions and by counseling patients to prevent treatment interruptions, and (e) collaborating with the multidisciplinary medical team to pursue imatinib dose escalation or other treatment options if patients have primary or acquired mutation-based resistance to imatinib.


CCL2 and CCR2 regulate pain-related behaviour and early gene expression in post-traumatic murine osteoarthritis but contribute little to chondropathy.
The role of inflammation in structural and symptomatic osteoarthritis (OA) remains unclear. One key mediator of inflammation is the chemokine CCL2, primarily responsible for attracting monocytes to sites of injury. We investigated the role of CCL2 and its receptor CCR2 in experimental OA. OA was induced in 10 weeks old male wild type (WT), Ccl2-/- and Ccr2-/- mice, by destabilisation of the medial meniscus (DMM). RNA was extracted from whole joints at 6 h and 7 days post-surgery and examined by reverse transcription polymerase chain reaction (RT-PCR). Gene expression changes between naïve and DMM-operated mice were compared. Chondropathy scores, from mice at 8, 12, 16 and 20 weeks post DMM were calculated using modified Osteoarthritis Research Society International (OARSI) grading systems. Changes in hind paw weight distribution, as a measure of pain, were assessed by Linton incapacitance. Absence of CCL2 strongly suppressed (>90%) selective inflammatory response genes in the joint 6 h post DMM, including arginase 1, prostaglandin synthase 2, nitric oxide synthase 2 and inhibin A. IL6, MMP3 and tissue inhibitor of metalloproteinase 1 were also significantly suppressed. Similar trends were also observed in the absence of CCR2. A lower average chondropathy score was observed in both Ccl2-/- and Ccr2-/- mice at 12, 16 and 20 weeks post DMM compared with WT mice, but this was only statistically significant at 20 weeks in Ccr2-/- mice. Pain-related behaviour in Ccl2-/- and Ccr2-/- mice post DMM was delayed in onset. The CCL2/CCR2 axis plays an important role in the development of pain in murine OA, but contributes little to cartilage damage.


GpIbα interacts exclusively with exosite II of thrombin.
Activation of platelets by the serine protease thrombin is a critical event in haemostasis. This process involves the binding of thrombin to glycoprotein Ibα (GpIbα) and cleavage of protease-activated receptors (PARs). The N-terminal extracellular domain of GpIbα contains an acidic peptide stretch that has been identified as the main thrombin binding site, and both anion binding exosites of thrombin have been implicated in GpIbα binding, but it remains unclear how they are involved. This issue is of critical importance for the mechanism of platelet activation by thrombin. If both exosites bind to GpIbα, thrombin could potentially act as a platelet adhesion molecule or receptor dimerisation trigger. Alternatively, if only a single site is involved, GpIbα may serve as a cofactor for PAR-1 activation by thrombin. To determine the involvement of thrombin's two exosites in GpIbα binding, we employed the complementary methods of mutational analysis, binding studies, X-ray crystallography and NMR spectroscopy. Our results indicate that the peptide corresponding to the C-terminal portion of GpIbα and the entire extracellular domain bind exclusively to thrombin's exosite II. The interaction of thrombin with GpIbα thus serves to recruit thrombin activity to the platelet surface while leaving exosite I free for PAR-1 recognition.


Anodal block permits directional vagus nerve stimulation.
Vagus nerve stimulation (VNS) is a bioelectronic therapy for disorders of the brain and peripheral organs, and a tool to study the physiology of autonomic circuits. Selective activation of afferent or efferent vagal fibers can maximize efficacy and minimize off-target effects of VNS. Anodal block (ABL) has been used to achieve directional fiber activation in nerve stimulation. However, evidence for directional VNS with ABL has been scarce and inconsistent, and it is unknown whether ABL permits directional fiber activation with respect to functional effects of VNS. Through a series of vagotomies, we established physiological markers for afferent and efferent fiber activation by VNS: stimulus-elicited change in breathing rate (ΔBR) and heart rate (ΔHR), respectively. Bipolar VNS trains of both polarities elicited mixed ΔHR and ΔBR responses. Cathode cephalad polarity caused an afferent pattern of responses (relatively stronger ΔBR) whereas cathode caudad caused an efferent pattern (stronger ΔHR). Additionally, left VNS elicited a greater afferent and right VNS a greater efferent response. By analyzing stimulus-evoked compound nerve potentials, we confirmed that such polarity differences in functional responses to VNS can be explained by ABL of A- and B-fiber activation. We conclude that ABL is a mechanism that can be leveraged for directional VNS.


Liposome surface functionalization based on different anchoring lipids via Staudinger ligation.
Liposome surface functionalization facilitates numerous potential applications of liposomes, such as enhanced stability, bioactive liposome conjugates, and targeted drug, gene and image agent delivery. Anchoring lipids are needed for grafting ligands of interest and play important roles in ligand grafting density, liposome stability, and liposome chemical and physical characteristics as well. In this report, glyco-functionalized liposome systems based on two kinds of anchoring lipids, phosphatidylethanolamine (PE) and cholesterol (Chol), were prepared by post chemically selective functionalization via Staudinger ligation. The size and stability of the liposomes were confirmed by dynamic light scattering (DLS). Particularly, the impact of anchor lipids on the stability of glyco-functionalized liposomes was investigated by comparing two different anchor lipids, namely Chol-PEG2000-TP and DSPE-PEG2000-TP. In addition, the encapsulation and releasing capacity of the glycosylated liposome based on the two anchoring lipids were investigated by entrapping 5,6-carboxyfluorescein (CF) dye and monitoring the fluorescence leakage, respectively. Furthermore, the density and accessibility of grafted carbohydrate residues on the liposome surface were evaluated for the two anchoring lipid-derived liposomes with lectin binding, respectively.


Arginase 2 deficiency prevents oxidative stress and limits hyperoxia-induced retinal vascular degeneration.
Hyperoxia exposure of premature infants causes obliteration of the immature retinal microvessels, leading to a condition of proliferative vitreoretinal neovascularization termed retinopathy of prematurity (ROP). Previous work has demonstrated that the hyperoxia-induced vascular injury is mediated by dysfunction of endothelial nitric oxide synthase resulting in peroxynitrite formation. This study was undertaken to determine the involvement of the ureahydrolase enzyme arginase in this pathology. Studies were performed using hyperoxia-treated bovine retinal endothelial cells (BRE) and mice with oxygen-induced retinopathy (OIR) as experimental models of ROP. Treatment with the specific arginase inhibitor 2(S)-amino-6-boronohexanoic acid (ABH) prevented hyperoxia-induced apoptosis of BRE cells and reduced vaso-obliteration in the OIR model. Furthermore, deletion of the arginase 2 gene protected against hyperoxia-induced vaso-obliteration, enhanced physiological vascular repair, and reduced retinal neovascularization in the OIR model. Additional deletion of one copy of arginase 1 did not improve the vascular pathology. Analyses of peroxynitrite by quantitation of its biomarker nitrotyrosine, superoxide by dihydroethidium imaging and NO formation by diaminofluoroscein imaging showed that the protective actions of arginase 2 deletion were associated with blockade of superoxide and peroxynitrite formation and normalization of NOS activity. Our data demonstrate the involvement of arginase activity and arginase 2 expression in hyperoxia-induced vascular injury. Arginase 2 deletion prevents hyperoxia-induced retinal vascular injury by preventing NOS uncoupling resulting in decreased reactive oxygen species formation and increased nitric oxide bioavailability.


Objective Assessment of Patient Inhaler User Technique Using an Audio-Based Classification Approach.
Many patients make critical user technique errors when using pressurised metered dose inhalers (pMDIs) which reduce the clinical efficacy of respiratory medication. Such critical errors include poor actuation coordination (poor timing of medication release during inhalation) and inhaling too fast (peak inspiratory flow rate over 90 L/min). Here, we present a novel audio-based method that objectively assesses patient pMDI user technique. The Inhaler Compliance Assessment device was employed to record inhaler audio signals from 62 respiratory patients as they used a pMDI with an In-Check Flo-Tone device attached to the inhaler mouthpiece. Using a quadratic discriminant analysis approach, the audio-based method generated a total frame-by-frame accuracy of 88.2% in classifying sound events (actuation, inhalation and exhalation). The audio-based method estimated the peak inspiratory flow rate and volume of inhalations with an accuracy of 88.2% and 83.94% respectively. It was detected that 89% of patients made at least one critical user technique error even after tuition from an expert clinical reviewer. This method provides a more clinically accurate assessment of patient inhaler user technique than standard checklist methods.


Organellar proteomics reveals hundreds of novel nuclear proteins in the malaria parasite Plasmodium falciparum.
The post-genomic era of malaria research provided unprecedented insights into the biology of Plasmodium parasites. Due to the large evolutionary distance to model eukaryotes, however, we lack a profound understanding of many processes in Plasmodium biology. One example is the cell nucleus, which controls the parasite genome in a development- and cell cycle-specific manner through mostly unknown mechanisms. To study this important organelle in detail, we conducted an integrative analysis of the P. falciparum nuclear proteome. We combined high accuracy mass spectrometry and bioinformatic approaches to present for the first time an experimentally determined core nuclear proteome for P. falciparum. Besides a large number of factors implicated in known nuclear processes, one-third of all detected proteins carry no functional annotation, including many phylum- or genus-specific factors. Importantly, extensive experimental validation using 30 transgenic cell lines confirmed the high specificity of this inventory, and revealed distinct nuclear localization patterns of hitherto uncharacterized proteins. Further, our detailed analysis identified novel protein domains potentially implicated in gene transcription pathways, and sheds important new light on nuclear compartments and processes including regulatory complexes, the nucleolus, nuclear pores, and nuclear import pathways. Our study provides comprehensive new insight into the biology of the Plasmodium nucleus and will serve as an important platform for dissecting general and parasite-specific nuclear processes in malaria parasites. Moreover, as the first nuclear proteome characterized in any protist organism, it will provide an important resource for studying evolutionary aspects of nuclear biology.


Ethanol-induced changes in the content of thiol compounds and of lipid peroxidation in livers and brains from mice: protection by thiazolidine derivatives.
Treatment of mice with ethanol for 5 days resulted in a drop of total glutathione in the liver, possibly due to an ethanol-stimulated increased conversion into L-cysteine. The levels of L-cysteine and the rate of lipid peroxidation were above control levels. Similar but less pronounced changes were observed with brain tissue. The continuation of the treatment with ethanol led to an adaptation in both tissues as assessed at days 10 and 15. These findings suggest induction of enzymes involved in the defence mechanisms against lipid peroxidation. However, at day 23 of treatment the levels of total glutathione and L-cysteine were reduced in the liver whereas lipid peroxidation was increased. Thus, a state of impaired defence mechanisms occurred during prolonged treatment. Interestingly, the concentration of total glutathione was increased in the brain suggesting protective mechanisms in this organ and possibly a supply from other organs. No increase of lipid peroxidation levels in the brain was observed. The substitution of the deficit of thiol compounds is a major problem because neither L-cysteine nor glutathione can be utilized for different reasons. Therefore, we treated mice with thiazolidine derivatives which can be regarded as 'frozen' L-cysteine. Two days of treatment with 2-methyl-thiazolidine-2,4-dicarboxylic acid were sufficient to observe an increase of total glutathione and free L-cysteine levels and a decrease of lipid peroxidation in the liver. These findings demonstrate a new treatment for the tissue-damaging effects of chronic ethanol ingestion.(ABSTRACT TRUNCATED AT 250 WORDS)


Localization of the activin signal transduction components in normal human ovarian follicles: implications for autocrine and paracrine signaling in the ovary.
The intraovarian function of gonadally produced activin is unclear, and many in vitro studies have suggested a role for activin in follicle development. To identify the follicular developmental stages at which these ligands may be acting, we have used immunohistochemical localization of the ligand subunits, receptor subtypes, and Smad co-activating proteins within the same follicles. The earliest stages of follicle development (primordial to primary) show no immunoreactivity for the activin subunits or their receptors. Oocytes from these early stages contain immunostaining for Smad2 and Smad4, consistent with signaling by other TGF-beta superfamily members. Immunostaining for the activin type II receptor first appears in oocytes and oocyte-associated cumulus cells at the secondary follicle stage. However, activin is not produced in these follicles, suggesting that either the receptors are inactive at this stage or they are used by another protein. Co-localization of activin and inhibin subunits, receptors, and Smads only occurs in granulosa and theca cells of small antral, aromatase-positive follicles as well as granulosa cells of early atretic follicles. In addition, multivariate statistical analysis reveals that the ligands and their cellular signaling complexes are independently regulated. Together, these data strongly suggest that the intraovarian role of activin is limited to a few developmental stages and that other TGF-beta family members may use this cell autonomous signaling machinery in early follicle development.


Fidelity of metal insertion into hydrogenases.
The fidelity of metal incorporation into the active center of hydrogenase 3 from Escherichia coli was studied by analyzing the inhibition of the maturation pathway by zinc and other transition metals. Hydrogenase maturation of wild-type cells was significantly affected only by concentrations of zinc or cadmium higher than 200 microM, whereas a mutant with a lesion in the nickel uptake system displayed a total blockade of the proteolytic processing of the precursor form into the mature form of the large subunit after growth in the presence of 10 microM Zn(2+). The precursor could not be processed in vitro by the maturation endopeptidase even in the presence of an excess of nickel ions. Evidence is presented that zinc does not interfere with the incorporation of iron into the metal center. Precursor of the large subunit accumulated in nickel proficient cells formed a transient substrate complex with the cognate endoprotease HycI whereas that of zinc-supplemented cells did not. The results show that zinc can intrude the nickel-dependent maturation pathway only when nickel uptake is blocked. Under this condition zinc appears to be incorporated at the nickel site of the large subunit and delivers a precursor not amenable to proteolytic processing since the interaction with the endoprotease is blocked.


Understanding emergency department staff needs and perceptions in the provision of palliative care.
The primary aim of the present study was to investigate Australian ED staff perspectives and needs regarding palliative care provision. Secondary aims were to assess staff views about death and dying, and their awareness of common causes of death in Australia, particularly those where a palliative care approach is appropriate. All medical and nursing staff working in a private ED in Perth, Western Australia, were asked to complete a combined quantitative and qualitative survey. The survey tool uses a combination of Likert-type scales and open-ended questions. Descriptive statistics and intergroup comparisons were made for all quantifiable variables, whereas formal content analysis was used for text responses. Surveys were returned by 22 doctors and 44 nurses, with most reporting only working knowledge of palliative care but clinical proficiency in symptom control. Confidence in palliative care provision was lower among nursing than medical staff but educational needs were similar. Cancer diagnoses were consistently overestimated, and dementia and COPD underestimated, as the most common causes of death. Only six of 63 (9.5%) of respondents identified the correct top five causes of death. Our study suggests that although ED staff expressed confidence regarding symptom management in palliative care, they lacked understanding of the patients in whom a palliative approach could be applied and sought further education in areas, such as end-of-life communication and ethical issues. ED specific training and clinical interventions in palliative care provision would seem to be needed and justified.


Influence of hapten density on immunogenicity for anti-ciprofloxacin antibody production in mice.
To generate antibodies against small molecules, it is necessary to couple them as haptens to large carriers such as proteins. However, the immunogenicity of the conjugates usually has no linear correlation with the hapten-protein ratio, which may lead to large variations in the character of the desired antibodies. In the present study, ciprofloxacin (CPFX) was coupled to bovine serum albumin (BSA) in five different proportions using a modified carbodiimide method. The conjugates were characterized qualitatively by spectrophotometric absorption and electrophoresis methods. Mass spectrometry and the trinitrobenzene sulfonic acid method were adopted to assay the density of conjugates quantitatively. As a result, CPFX-BSA conjugates with various hapten densities (21-30 molecules per carrier protein) were obtained. After immunization in mice, ELISA tests showed that the antisera titer increased gradually with the increase of hapten density. The antibody obtained from the mice showed high sensitivity toward CPFX. These results revealed the relationship between hapten density and immunogenicity as well as an optimized conjugation approach for immunization purposes.


Cheese: Food Perception and Food Choice.
In light of the increasing interest in the economic and socio-political impact of the 'traditional food' trend, it is essential to understand the determinant factors that lead to traditional consumer choices. The standardization of sensory quality evaluation methods marks the pressing need for food product certification, particularly foods with specific sensory characteristics, such as those with a Protected Designation of Origin (PDO). Consumer perception of particular foods, especially for foods that are culturally and socially contingent, such as cheese, must be understood as both a psychophysical reflex and a learned social practice. Consumers create their own perceptions based on the overall intrinsic or extrinsic cheese characteristics, mainly sensory characteristics that reflect others' attributes. These characteristics are normally linked to the specific cheese manufacture process. Some patents propose the use of adapted cheesemaking equipment (EP1982582A2), suitable for the manufacture of small-scale cheeses, such as some PDO cheese. Thus, sensory evaluation of any kind of cheese is based, in the initial phase, on knowledge of the sensory methods for cheese evaluation and, in a second phase, on the familiarity of the cheese characteristics and verbalization of desirable and undesirable attributes. This paper presents a case study based on the traditional food product, Évora cheese, assembled with PDO cheeses, whose sensory and physicochemical quality attributes are essential in order to obtain this designation and ensure the genuine properties that characterize them, as well as ascertaining exactly how they are perceived and further accepted by the consumer.


Comparisons of Noninvasive Methods Used to Assess Exercise Stroke Volume in Heart Failure with Preserved Ejection Fraction.
Cardiopulmonary exercise testing (CPET) plays an important role in properly phenotyping signs and symptoms of heart failure with preserved ejection fraction (HFpEF). The prognostic value of CPET is strengthened when accompanied by cardiac hemodynamic measurements. Although recognized as the "gold" standard, cardiac catheterization is impractical for routine CPET. Thus, advancing the scientific/methodologic understanding of noninvasive techniques for exercise cardiac hemodynamic assessment is clinically impactful in HFpEF. This study tested the concurrent validity of noninvasive acetylene gas (C2H2) uptake, echocardiography (ECHO), and oxygen pulse (O2pulse) for measuring/predicting exercise stroke volume (SV) in HFpEF. Eighteen white HFpEF and 18 age-/sex-matched healthy controls participated in upright CPET (ages, 69 ± 9 yr vs 63 ± 9 yr). At rest, 20 W, and peak exercise, SV was measured at steady-state via C2H2 rebreathe (SVACET) and ECHO (SVECHO), whereas O2pulse was derived (=V˙O2/HR). Resting relationships between SVACET and SVECHO, SVECHO and O2pulse, or SVACET and O2pulse were significant in HFpEF (R = 0.30, 0.36, 0.67), but not controls (R = 0.07, 0.01, 0.09), respectively. Resting relationships persisted to 20 W in HFpEF (R = 0.70, 0.53, 0.70) and controls (R = 0.05, 0.07, 0.21), respectively. Peak exercise relationships were significant in HFpEF (R = 0.62, 0.24, 0.64), but only for SVACET versus O2pulse in controls (R = 0.07, 0.04, 0.33), respectively. Standardized standard error of estimate between techniques was strongest in HFpEF at 20 W: SVACET versus SVECHO = 0.57 ± 0.22; SVECHO versus O2pulse = 0.71 ± 0.28; SVACET versus O2pulse = 0.56 ± 0.22. Constituting a clinically impactful step towards construct validation testing, these data suggest SVACET, SVECHO, and O2pulse demonstrate moderate-to-strong concurrent validity for measuring/predicting exercise SV in HFpEF.


Differential effects of mindful breathing, progressive muscle relaxation, and loving-kindness meditation on decentering and negative reactions to repetitive thoughts.
Decentering has been proposed as a potential mechanism of mindfulness-based interventions but has received limited empirical examination to date in experimental studies comparing mindfulness meditation to active comparison conditions. In the present study, we compared the immediate effects of mindful breathing (MB) to two alternative stress-management techniques: progressive muscle relaxation (PMR) and loving-kindness meditation (LKM) to test whether decentering is unique to mindfulness meditation or common across approaches. Novice meditators (190 female undergraduates) were randomly assigned to complete one of three 15-min stress-management exercises (MB, PMR, or LKM) presented by audio recording. Immediately after the exercise, participants completed measures of decentering, frequency of repetitive thoughts during the exercise, and degree of negative reaction to thoughts. As predicted, participants in the MB condition reported greater decentering relative to the other two conditions. The association between frequency of repetitive thought and negative reactions to thoughts was relatively weaker in the MB condition than in the PMR and LKM conditions, in which these two variables were strongly and positively correlated. Consistent with the construct of decentering, the relative independence between these two variables in the MB condition suggests that mindful breathing may help to reduce reactivity to repetitive thoughts. Taken together, results help to provide further evidence of decentering as a potential mechanism that distinguishes mindfulness practice from other credible stress-management approaches.


Risk factors of post endoscopic retrograde cholangiopancreatography bacteremia.
Bacteremia following endoscopic retrograde cholangiopancreatography (ERCP) is a severe complication, but the risk factors for this condition have not yet been clearly determined. Thus, the aim of this study was to investigate the risk factors of post-ERCP bacteremia. Among patients who underwent ERCP from June 2006 to May 2009, we selected patients without any signs of infection prior to the ERCP procedures. Of these patients, we further selected those who experienced bacteremia after ERCP as well as two-fold age and sex-matched controls who did not experience bacteremia after ERCP procedures. We compared clinical, laboratory and technical aspects between these two groups. There were 70 patients (3.1%) who developed bacteremia after ERCP. In the multivariate analysis, a history of previous liver transplantation, an elevated serum alkaline phosphatase level and an endoscopic retrograde biliary drainage procedure were independent risk factors of post-ERCP bacteremia (p=0.006, p=0.001, and p=0.004, respectively). The microbiologic analysis revealed the presence of gram-negative organisms in 80% of the cases, and 11 patients had infections with bacteria expressing extended spectrum β-lactamases. Pseudomonas infection was significantly more common in patients who received liver transplantation as compared to patients without transplantation (p=0.014). A history of liver transplantation, elevated serum alkaline phosphatase levels and endoscopic retrograde biliary drainage procedure were independent risk factors of post-ERCP bacteremia and require additional attention in future studies.


Commelinaceomyces, gen. nov., for four clavicipitaceous species misplaced in Ustilago that infect Commelinaceae.
A fungus causing false smut in the flowers of Murdannia keisak (Commelinaceae, Commelinales, Monocots) in Japan was morphologically identical to Ustilago aneilematis. The fungus infected ovaries of most flowers of host plants. Infected flowers were filled with yellow to orange thick-walled conidia that became olivaceous green at maturity. However, multilocus phylogenetic analysis of DNA sequences (18S, 28S, translation elongation factor 1α [TEF], the largest [RPB1] and the second largest [RPB2] subunit of RNA polymerase II) showed that the fungus belonged to the tribe Ustilaginoideae (Clavicipitaceae, Hypocreales, Ascomycota). Microscopic examination showed that the fungus developed conidia at the apex of conidiogenous cells, in contrast to other species in the Ustilaginoideae that develop conidia pleurogenously. A new genus, Commelinaceomyces, is formally proposed in the Ustilaginoideae to accommodate this fungus. Four species previously misplaced in Ustilago (Ustilaginales, Basidiomycota) are transferred to Commelinaceomyces, including the type of the genus, C. aneilematis, on Murdannia keisak. This is the first report of a clavicipitaceous species infecting host plants in the Commelinaceae.


Daclatasvir plus sofosbuvir for previously treated or untreated chronic HCV infection.
All-oral combination therapy is desirable for patients with chronic hepatitis C virus (HCV) infection. We evaluated daclatasvir (an HCV NS5A replication complex inhibitor) plus sofosbuvir (a nucleotide analogue HCV NS5B polymerase inhibitor) in patients infected with HCV genotype 1, 2, or 3. In this open-label study, we initially randomly assigned 44 previously untreated patients with HCV genotype 1 infection and 44 patients infected with HCV genotype 2 or 3 to daclatasvir at a dose of 60 mg orally once daily plus sofosbuvir at a dose of 400 mg orally once daily, with or without ribavirin, for 24 weeks. The study was expanded to include 123 additional patients with genotype 1 infection who were randomly assigned to daclatasvir plus sofosbuvir, with or without ribavirin, for 12 weeks (82 previously untreated patients) or 24 weeks (41 patients who had previous virologic failure with telaprevir or boceprevir plus peginterferon alfa-ribavirin). The primary end point was a sustained virologic response (an HCV RNA level of <25 IU per milliliter) at week 12 after the end of therapy. Overall, 211 patients received treatment. Among patients with genotype 1 infection, 98% of 126 previously untreated patients and 98% of 41 patients who did not have a sustained virologic response with HCV protease inhibitors had a sustained virologic response at week 12 after the end of therapy. A total of 92% of 26 patients with genotype 2 infection and 89% of 18 patients with genotype 3 infection had a sustained virologic response at week 12. High rates of sustained virologic response at week 12 were observed among patients with HCV subtypes 1a and 1b (98% and 100%, respectively) and those with CC and non-CC IL28B genotypes (93% and 98%, respectively), as well as among patients who received ribavirin and those who did not (94% and 98%, respectively). The most common adverse events were fatigue, headache, and nausea. Once-daily oral daclatasvir plus sofosbuvir was associated with high rates of sustained virologic response among patients infected with HCV genotype 1, 2, or 3, including patients with no response to prior therapy with telaprevir or boceprevir. (Funded by Bristol-Myers Squibb and Pharmasset (Gilead); A1444040 ClinicalTrials.gov number, NCT01359644.).


Asymmetric aza-Henry reaction under phase transfer catalysis: an experimental and theoretical study.
An efficient catalytic asymmetric aza-Henry reaction under phase transfer conditions is presented. The method is based on the reaction of the respective nitroalkane with alpha-amido sulfones effected by CsOH x H2O base in toluene as solvent and in the presence of cinchone-derived ammonium catalysts. This direct aza-Henry reaction presents as interesting features its validity for both nonenolizable and enolizable aldehyde-derived azomethines and the tolerance of nitroalkanes, other than nitromethane, for the production of beta-nitroamines. The synthetic value of the methodology described is demonstrated by providing (a) a direct route for the asymmetric synthesis of differently substituted 1,2-diamines and (b) a new asymmetric synthesis of gamma-amino alpha,beta-unsaturated esters through a catalytic, highly enantioselective formal addition of functionalized alkenyl groups to azomethines. Finally, a preferred TS that nicely fits the observed enantioselectivity has been identified. Most remarkable, an unusual hydrogen bond pattern for the catalyst-nitrocompound-imine complex is predicted, where the catalyst OH group interacts with the NO2 group of the nitrocompound.


Longitudinal Association Between Physical Activity and Body Fat During Adolescence: A Systematic Review.
Physical activity (PA) practice has been inversely associated to body fat (BF) and recommended as a way to reduce and prevent obesity. The objective of this study was to conduct a systematic review on the association of PA and BF in adolescence. The review includes 18 longitudinal studies found in the PubMed database, comprising papers published from January 1990 to July 2014. Studies assessing BF only through body mass index were excluded. Among the outcomes analyzed, waist circumference, skinfolds, and absolute and relative fat mass measurement were identified. Questionnaires were the more predominant way to evaluate PA. Most studies showed that PA promotes a protective effect against a higher BF gain. It was concluded that PA has a protective effect against BF with differences between the genders and according to the BF marker or measurement assessed; higher intensity PA leads to a greater effect against BF gain in both genders; and the maintenance or increase of PA level on BF observed through analysis of change in PA level yielded more consistent findings in the relation between PA and BF.


Development of the Alcohol Quality of Life Scale (AQoLS): a new patient-reported outcome measure to assess health-related quality of life in alcohol use disorder.
The development of patient-centred approaches and of reduction in consumption strategies in alcohol use disorder requires giving a larger place to qualitative assessments that are closer to patients' concerns and more clinically relevant than drinking outcomes and generic health-related quality of life instrument. Our purpose was to develop from patients input the Alcohol Quality of Life Scale (AQoLS), a disease-specific measure for alcohol use disorder (AUD). Concept elicitation focus groups were conducted with AUD patients in the UK and France. Thematic analysis was used to identify key areas of impact of AUD, and draft items were developed to capture these issues. The draft items underwent expert review to ensure clinical and cross-cultural applicability. Two iterative rounds of cognitive debriefing interviews were conducted with AUD patients in both countries, to assess face and content validity. From focus groups conducted with 38 AUD patients, seven areas of impact emerged, forming the basis for the AQoLS: relationships, activities, looking after self, emotional impact, control, living conditions, and sleep. An initial pool of 90 items was reduced to 50 following the review process. In cognitive interviews, the measure took less than 10 min to complete, and patients reported that items were relevant to their experience. Following Round 1 interviews (n = 16), 14 items were removed because patients felt they were unclear or uneasy to respond to, 2 were combined, 7 were revised, and 4 new items were added. The recall period of 2 weeks was changed to 4 weeks, based on patient comments. Following Round 2 interviews (n = 15), 5 items were removed and 3 were modified to produce the 34-item AQoLS. As the only de novo measure of health-related quality of life developed specifically for AUD, the AQoLS offers the potential of increased sensitivity to show the effectiveness of therapeutic interventions from the patient's perspective.


Transient ictal psychosis.
Transient psychotic episodes may result from continuous cerebral epileptiform discharges unaccompanied by clinically observed seizures. Such episodes may mimic depressive, hysterical and schizophrenic psychosis and delirium. I describe two patients here and review eight patient histories from the literature. Diagnosis is established by use of electroencephalography during the psychotic episode. Correct diagnosis is essential because specific treatment is available and additional episodes may be prevented by appropriate long-term anticonvulsant therapy.


Value of negative spiral CT angiography in patients with suspected acute PE: analysis of PE occurrence and outcome.
The aim of this study was to analyze pulmonary embolism (PE) occurrence and retrospective clinical outcome in patients with clinically suspected acute PE and a negative spiral CT angiography (SCTA) of the pulmonary arteries. Within a 35-month period, 485 consecutive patients with clinical symptoms of acute PE underwent SCTA of the pulmonary arteries. Patients with a negative SCTA and without anticoagulation treatment were followed-up and formed the study group. Patient outcome and recurrence of PE was evaluated retrospectively during a period of 6 months after the initial SCTA, and included a review of computerized patient records, and interviews with physicians and patients. Patients were asked to fill out a questionnaire concerning all relevant questions about their medical history and clinical course during the follow-up period. Special attention was focused on symptoms indicating recurrent PE, as well as later confirmation and therapy of PE. Of the 485 patients, 325 patients (67%) had a negative scan, 134 (27.6%) had radiological signs of PE, and 26 (5.4%) had an indeterminant result. Of 325 patients with a negative scan, 269 (83%) were available for follow-up. The main reasons for loss to follow-up were change of address, name, or phone number, or non-resident patients who left abroad. Of 269 patients available for follow-up, 49 patients (18.2% of 269) received anticoagulant treatment because of prior or recent deep venous thrombosis (32.6%) or a history of PE (34.7%), cardiovascular disease (18.4%), high clinical probability (8.2%), positive ventilation-perfusion scan (4.2%), and elevated D-dimer test (2%). The remaining 220 patients, who did not receive anticoagulant medication, formed the study group. Of this study group, 1 patient died from myocardial infarction 6 weeks after the initial SCTA, and the postmortem examination also detected multiple peripheral emboli in both lungs ( p=0.45%; 0.01-2.5, 95% confidence interval). The PE did not occur in any other patient. In patients with suspected PE and negative SCTA without anticoagulant therapy, the risk of recurrent PE in this study was less than 1% and similar to that in patients after a negative pulmonary angiogram. Therefore, we conclude that patients can be managed safely without anticoagulation therapy; however, this approach may not be appropriate for critically ill patients and those with persistent high clinical suspicion of acute PE.


Binary pattern deflectometry.
Deflectometry is widely used to accurately calculate the slopes of any specular reflective surface, ranging from car bodies to nanometer-level mirrors. This paper presents a new deflectometry technique using binary patterns of increasing frequency to retrieve the surface slopes. Binary Pattern Deflectometry allows almost instant, simple, and accurate slope retrieval, which is required for applications using mobile devices. The paper details the theory of this deflectometry method and the challenges of its implementation. Furthermore, the binary pattern method can also be combined with a classic phase-shifting method to eliminate the need of a complex unwrapping algorithm and retrieve the absolute phase, especially in cases like segmented optics, where spatial algorithms have difficulties. Finally, whether it is used as a stand-alone or combined with phase-shifting, the binary patterns can, within seconds, calculate the slopes of any specular reflective surface.


Selected Food Consumption Mediates the Association between Education Level and Metabolic Syndrome in Korean Adults.
Low socioeconomic status (SES) is linked to higher incidence/mortality of cardiovascular disease, but emerging evidence inconsistently reported that education level, a proxy for SES, is related to cardiovascular risk and metabolic syndrome (MetS) in Koreans. Furthermore, limited information is available on whether dietary components would mediate the relationship between education level and cardiovascular risk. We hypothesized that selected food consumption mediates the association between education level and MetS prevalence. Data from the Korea National Health and Nutritional Examination Survey (2008-2011) were included in cross-sectional analyses (n = 11,029, 30-64 years). The possible mediating effect of selected food groups (fruits, raw vegetables, red meat, milk, and soft drinks) on the association between education level and MetS was tested using a multiple mediation model. Education level was negatively associated with MetS prevalence. The association between lower education level and higher MetS prevalence was partially mediated by selected food consumption (lower intakes of fruit, red meat and milk; higher intakes of vegetable and soft drink) after adjusted for covariates. Gender also modified the association between education level and MetS prevalence that was more prominent in women than in men. Selected food consumption substantially contributes to the association between education level and MetS in Korean adults, especially among women.


Investigation into an outbreak of encephalomyelitis caused by a neuroinvasive porcine sapelovirus in the United Kingdom.
An outbreak of neurological disease in grower pigs characterised by ataxia and paraparesis was investigated in this study. The outbreak occurred 3-4 weeks post weaning in grower pigs which displayed signs of spinal cord damage progressing to recumbency. Pathology in the affected spinal cords and to a lesser extent in the brainstem was characterised by pronounced inflammation and neuronophagia in the grey matter. Molecular investigation using a pan-virus microarray identified a virus related to porcine sapelovirus (PSV) in the spinal cord of the two affected pigs examined. Analysis of 802 nucleotides of the virus polymerase gene showed the highest homology with those of viruses in the genus Sapelovirus of Picornaviridae. This PSV, strain G5, shared 91-93%, 67-69% and 63% nucleotide homology with porcine, simian and avian sapeloviruses, respectively. The nucleotide homology to other members of the Picornaviridae ranged from 41% to 62%. Furthermore, viral antigen was detected and co-localised in the spinal cord lesions of affected animals by an antibody known to react with PSV. In conclusion, clinical and laboratory observations of the diseased pigs in this outbreak are consistent with PSV-G5 being the causative agent. To the best of the authors' knowledge, this is the first unequivocal report of polioencephalomyelitis in pigs by a neuroinvasive PSV in the United Kingdom.


Nanoscale Near-Field Tomography of Surface States on (Bi0.5Sb0.5)2Te3.
Three-dimensional topological insulators (TIs) have attracted tremendous interest for their possibility to host massless Dirac Fermions in topologically protected surface states (TSSs), which may enable new kinds of high-speed electronics. However, recent reports have outlined the importance of band bending effects within these materials, which results in an additional two-dimensional electron gas (2DEG) with finite mass at the surface. TI surfaces are also known to be highly inhomogeneous on the nanoscale, which is masked in conventional far-field studies. Here, we use near-field microscopy in the mid-infrared spectral range to probe the local surface properties of custom-tailored (Bi0.5Sb0.5)2Te3 structures with nanometer precision in all three spatial dimensions. Applying nanotomography and nanospectroscopy, we reveal a few-nanometer-thick layer of high surface conductivity and retrieve its local dielectric function without assuming any model for the spectral response. This allows us to directly distinguish between different types of surface states. An intersubband transition within the massive 2DEG formed by quantum confinement in the bent conduction band manifests itself as a sharp, surface-bound, Lorentzian-shaped resonance. An additional broadband background in the imaginary part of the dielectric function may be caused by the TSS. Tracing the intersubband resonance with nanometer spatial precision, we observe changes of its frequency, likely originating from local variations of doping or/and the mixing ratio between Bi and Sb. Our results highlight the importance of studying the surfaces of these novel materials on the nanoscale to directly access the local optical and electronic properties via the dielectric function.


Effect of captivity on morphology: negligible changes in external morphology mask significant changes in internal morphology.
Captive breeding programmes are increasingly relied upon for threatened species management. Changes in morphology can occur in captivity, often with unknown consequences for reintroductions. Few studies have examined the morphological changes that occur in captive animals compared with wild animals. Further, the effect of multiple generations being maintained in captivity, and the potential effects of captivity on sexual dimorphism remain poorly understood. We compared external and internal morphology of captive and wild animals using house mouse (Mus musculus) as a model species. In addition, we looked at morphology across two captive generations, and compared morphology between sexes. We found no statistically significant differences in external morphology, but after one generation in captivity there was evidence for a shift in the internal morphology of captive-reared mice; captive-reared mice (two generations bred) had lighter combined kidney and spleen masses compared with wild-caught mice. Sexual dimorphism was maintained in captivity. Our findings demonstrate that captive breeding can alter internal morphology. Given that these morphological changes may impact organismal functioning and viability following release, further investigation is warranted. If the morphological change is shown to be maladaptive, these changes would have significant implications for captive-source populations that are used for reintroduction, including reduced survivorship.


Fracture Resistance of Immature Incisors Following Root Filling with Various Bioactive Endodontic Cements Using an Experimental Bovine Tooth Model.
The aim of this study was to compare the fracture resistance of immature bovine roots when using ProRoot MTA, CEM Cement, and Biodentine as root filling materials. An immature bovine tooth model was developed by removing the coronal and apical portions of 70 bovine incisors 8 mm above and 12 mm below the cementoenamel junction (CEJ). The specimens were then divided into five groups: ProRoot MTA, CEM Cement, Biodentine, gutta-percha/AH26 sealer, and control. All groups received a 5-mm apical plug with a temporary restorative material. Then, the remaining root canal space was filled with one of the afore-mentioned materials. After setting, the specimens were mounted in acrylic resin. Then, 3 mm coronal to the CEJ from the buccal side of the teeth and at a 135°angle to the long axis, the specimens were loaded until fracture. The specimens in the Biodentine (2196 N) and ProRoot MTA (2103 N) groups had significantly greater fracture resistance in comparison to the control group (p = 0.01). No significant difference was found between CEM Cement, gutta-percha and sealer AH26, and control groups. No significant differences occurred between the four experimental groups (p = 0.45). Filling the root canal space with ProRoot MTA and Biodentine contributed to higher fracture resistance values.


Three artificial intelligence data challenges based on CT and MRI.
The second edition of the artificial intelligence (AI) data challenge was organized by the French Society of Radiology with the aim to: (i), work on relevant public health issues; (ii), build large, multicentre, high quality databases; and (iii), include three-dimensional (3D) information and prognostic questions. Relevant clinical questions were proposed by French subspecialty colleges of radiology. Their feasibility was assessed by experts in the field of AI. A dedicated platform was set up for inclusion centers to safely upload their anonymized examinations in compliance with general data protection regulation. The quality of the database was checked by experts weekly with annotations performed by radiologists. Multidisciplinary teams competed between September 11th and October 13th 2019. Three questions were selected using different imaging and evaluation modalities, including: pulmonary nodule detection and classification from 3D computed tomography (CT), prediction of expanded disability status scale in multiple sclerosis using 3D magnetic resonance imaging (MRI) and segmentation of muscular surface for sarcopenia estimation from two-dimensional CT. A total of 4347 examinations were gathered of which only 6% were excluded. Three independent databases from 24 individual centers were created. A total of 143 participants were split into 20 multidisciplinary teams. Three data challenges with over 1200 general data protection regulation compliant CT or MRI examinations each were organized. Future challenges should be made with more complex situations combining histopathological or genetic information to resemble real life situations faced by radiologists in routine practice.


Net nutrient absorption and liver metabolism in lactating dairy cows fed supplemental dietary biotin.
The effect of feeding supplemental biotin on net absorption and metabolism of nutrients by the portal-drained viscera (PDV; the gut, pancreas, spleen and associated fat) and liver of lactating dairy cows was measured. Three cows in early to mid-lactation catheterised for measurements of net nutrient absorption and metabolism by the PDV and liver were fed a total-mixed ration with or without supplemental biotin at 20 mg/day using a switch-back design (ABA v. BAB) with three 2-week periods. There were no effects of feeding biotin on dry matter intake (22.2 kg/day), milk yield (29.5 kg/day) or milk composition. There was also no effect of feeding biotin on net release of glucose by the liver, net liver removal of glucose precursors (propionate, alanine, lactate) or net liver release of β-hydroxybutyrate. Feeding biotin increased net PDV release of ammonia. Reasons for the response are not certain, but a numerical increase in net PDV release of acetate suggests that rumen or hindgut fermentation was altered. Results of the present study do not support the hypothesis that supplemental biotin increases liver glucose production in lactating dairy cows.


Purification and properties of an extracellular blastogen produced by group A streptococci.
An extracellular product of group A streptococci which induces lymphocyte blastogenesis has been purified to homogeneity by DEAE-cellulose and CM-cellulose chromatography. The protein, termed streptococcal blastogen A, has a mol wt of approximately or equal to 17,500 and is inactivated by protease treatment and by heating at 100 degrees C. The purified blastogen gave rise to multiple protein bands on nondenaturing polyacrylamide gel electrophoresis, only two of which possessed blastogenic activity. Treatment of the protein with dithiothreitol before electrophoresis resulted in the apparent conversion of the multiple forms to a single active species. Blastogen A differs in electrophoretic mobility from the streptococcal pyrogenic exotoxins and its lymphocyte stimulating activity is not inhibited by rabbit antisera to the exotoxins. An enzyme immunoassay has been developed to measure human antibodies against blastogen A. A selection of sera with varying levels of anti-DNase B contained antiblastogen A-IgG.


Assessment of strain field in endothelial cells subjected to uniaxial deformation of their substrate.
A stretch chamber has been developed in order to visualize the deformation of cells subjected to controlled uniaxial stretch of their substrate. A rectangular, custom-made, transparent silicone channel is used as a deformable substrate. Bovine aortic endothelial cells are plated at the bottom of the channel whose lateral deformation is controlled by two piezoelectric translators. The system is mounted on the stage of a confocal microscope where three-dimensional (3D) images of the cells can be acquired simultaneously in the three RGB channels. The first channel provides images of 216 nm fluorescent beads embedded in the cytoskeleton (used as internal markers). The second is used to image the shape of the nucleus revealed by live cell nucleic acid staining. The third one provides a transmitted light image of the cell outline. 3D images of the cell are taken before deformation, after uniaxial deformation of the substrate (up to 25%) and after relaxation. Results indicate that: (a) the cell closely follows the deformation imposed by the substrate with no measurable residual strain after relaxation, and (b) there is a clear mechanical coupling between the extracellular matrix and the nucleus, which deforms significantly under the applied substrate stretch. Suggesting that the nucleus can directly sense the mechanical environment of the cell, the latter result has potentially important implications for signal transduction.


Live-cell Imaging with Genetically Encoded Protein Kinase Activity Reporters.
Protein kinases play pivotal roles in intracellular signal transduction, and dysregulation of kinases leads to pathological results such as malignant tumors. Kinase activity has hitherto been measured by biochemical methods such as in vitro phosphorylation assay and western blotting. However, these methods are less useful to explore spatial and temporal changes in kinase activity and its cell-to-cell variation. Recent advances in fluorescent proteins and live-cell imaging techniques enable us to visualize kinase activity in living cells with high spatial and temporal resolutions. Several genetically encoded kinase activity reporters, which are based on the modes of action of kinase activation and phosphorylation, are currently available. These reporters are classified into single-fluorophore kinase activity reporters and Förster (or fluorescence) resonance energy transfer (FRET)-based kinase activity reporters. Here, we introduce the principles of genetically encoded kinase activity reporters, and discuss the advantages and disadvantages of these reporters.Key words: kinase, FRET, phosphorylation, KTR.


Subcellular Organization of the cAMP Signaling Pathway.
The field of cAMP signaling is witnessing exciting developments with the recognition that cAMP is compartmentalized and that spatial regulation of cAMP is critical for faithful signal coding. This realization has changed our understanding of cAMP signaling from a model in which cAMP connects a receptor at the plasma membrane to an intracellular effector in a linear pathway to a model in which cAMP signals propagate within a complex network of alternative branches and the specific functional outcome strictly depends on local regulation of cAMP levels and on selective activation of a limited number of branches within the network. In this review, we cover some of the early studies and summarize more recent evidence supporting the model of compartmentalized cAMP signaling, and we discuss how this knowledge is starting to provide original mechanistic insight into cell physiology and a novel framework for the identification of disease mechanisms that potentially opens new avenues for therapeutic interventions. SIGNIFICANCE STATEMENT: cAMP mediates the intracellular response to multiple hormones and neurotransmitters. Signal fidelity and accurate coordination of a plethora of different cellular functions is achieved via organization of multiprotein signalosomes and cAMP compartmentalization in subcellular nanodomains. Defining the organization and regulation of subcellular cAMP nanocompartments is necessary if we want to understand the complex functional ramifications of pharmacological treatments that target G protein-coupled receptors and for generating a blueprint that can be used to develop precision medicine interventions.


Facilitating congruence between religious beliefs and sexual identity with mindfulness.
With the increasing relevance of sexual minority concerns, including the process of navigating sexual and religious identities, clinical practice has focused on helping sexual minorities address methods of self-expression that are most congruent with the client's values. Sexual Identity Therapy (SIT), (Throckmorton & Yarhouse, 2006) has been developed to assist individuals who are seeking to address potential conflicts between religious and sexual identities by focusing on personal congruence. To facilitate this process, the practice of mindfulness is applied. As an adaptation from its spiritual origins, mindfulness is used to facilitate the treatment of various disorders, such as chronic pain, substance abuse, and depression. It has also been the crux of several different third-wave cognitive and behavioral therapies that consider the "… context and functions of psychological phenomena" (Hayes, 2004, p. 5) for the purpose of helping clients to develop "… broad, flexible and effective repertoires" (p. 6). In this instance, mindfulness is applied to SIT to assist individuals with same-sex attraction to become nonjudgmentally aware of their thoughts and feelings related to same-sex attraction such that they are able to experience their attractions in an open and honest manner without feeling compelled to either dismiss or augment these attractions. Mindful awareness of same-sex attraction facilitates congruence because there is less emphasis on changing behaviors, thoughts or feelings, but rather, changing the relationship the individual has to their experiences of same-sex attraction so that they are experienced as neutral, as opposed to aversive.


Evidence for subclinical H5N1 avian influenza infections among Nigerian poultry workers.
In recent years Nigeria has experienced sporadic incursions of highly pathogenic H5N1 avian influenza among poultry. In 2008, 316 poultry-exposed agricultural workers, and 54 age-group matched non-poultry exposed adults living in the Enugu or Ebonyi States of Nigeria were enrolled and then contacted monthly for 24 months to identify acute influenza-like-illnesses. Annual follow-up sera and questionnaire data were collected at 12 and 24 months. Participants reporting influenza-like illness completed additional questionnaires, and provided nasal and pharyngeal swabs and acute and convalescent sera. Swab and sera specimens were studied for evidence of influenza A virus infection. Sera were examined for elevated antibodies against 12 avian influenza viruses by microneutralization and 3 human viruses by hemagglutination inhibition. Four (3.2%) of the 124 acute influenza-like-illness investigations yielded molecular evidence of influenza, but virus could not be cultured. Serial serum samples from five poultry-exposed subjects had a ≥4-fold change in microneutralization titers against A/CK/Nigeria/07/1132123(H5N1), with three of those having titers ≥1:80 (maximum 1:1,280). Three of the five subjects (60%) reported a preceding influenza-like illness. Hemagglutination inhibition titers were ≥4-fold increases against one of the human viruses in 260 participants. While cross-reactivity from antibodies against other influenza viruses cannot be ruled out as a partial confounder, over the course of the 2-year follow-up, at least 3 of 316 (0.9%) poultry-exposed subjects had evidence for subclinical HPAI H5N1 infections. If these data represent true infections, it seems imperative to increase monitoring for avian influenza among Nigeria's poultry and poultry workers.


Poly (A) Binding Protein Cytoplasmic 1 Is a Novel Co-Regulator of the Androgen Receptor.
The androgen receptor (AR) is a member of the steroid receptor superfamily that regulates gene expression in a ligand-dependent manner. The NTD of the AR plays a key role in AR transactivation including androgen-independent activation of the AR in castration-resistant prostate cancer (CRPC) cells. We recently reported that amino acids (a.a.) 50-250 of the NTD are capable of modulating AR nucleocytoplasmic trafficking. To further explore the mechanism associated with a.a. 50-250, GFP pull-down assays were performed in C4-2 CRPC cells transfected with GFP tagged a.a. 50-250 of the AR. Mass spectrometry analysis of the pulled down proteins identified poly (A) binding protein cytoplasmic 1 (PABPC1) interaction with this region of the AR. In silico analysis of gene expression data revealed PABPC1 up-regulation in prostate cancer tissue specimens and this up-regulation correlates to increased disease recurrence. Co-immunoprecipitation assays confirmed the association of PABPC1 with a.a. 50-250 of the NTD of the AR. Knockdown of PABPC1 decreased nuclear AR protein levels and inhibited androgen activation of the AR target PSA in LNCaP and C4-2 cells. Additionally, knockdown of PABPC1 inhibited transactivation of the PSA promoter by NAR (AR lacking the LBD) and attenuated proliferation of AR-positive prostate cancer cells. These findings suggest that PABPC1 is a novel co-regulator of the AR and may be a potential target for blocking activation of the AR in CRPC.


Droplet-based single cell RNAseq tools: a practical guide.
Droplet based scRNA-seq systems such as Drop-seq, inDrop and Chromium 10X have been the catalyst for the wide adoption of high-throughput scRNA-seq technologies in the research laboratory. In order to understand the capabilities of these systems to deeply interrogate biology; here we provide a practical guide through all the steps involved in a typical scRNA-seq experiment. Through comparing and contrasting these three main droplet based systems (and their derivatives), we provide an overview of all critical considerations in obtaining high quality and biologically relevant data. We also discuss the limitations of these systems and how they fit into the emerging field of Genomic Cytometry.


Spread of Rotylenchulus reniformis in an Arkansas Cotton Field Over a Four-Year Period.
Rotylenchulus reniformis was first detected in a single grid (100 m(2)) in May 2001 in a cotton field in Ashley County, AR, that was being utilized to evaluate the utility of grid-sampling for detection of Meloidogyne incognita. A total of 512 grids were sampled in the 6-ha field in the spring and fall for four years (2001 - 2004), nematode populations were determined for each grid, and nematode population density maps were constructed utilizing Global Positioning Systems and Geographic Information Systems. In May 2001, R. reniformis population density in the single grid where it was detected was 6,364 juveniles and adult reniform nematodes/500 cm(3) soil. By the end of the first year (October 2001), the nematode was found in 17 of the 512 plots with population densities ranging from 682 to 10,909 nematodes/500 cm(3) soil. Over the course of the 4-yr period, reniform nematode incidence increased to 107 of 512 plots, with population density ranging from 227 to 32,727 nematodes/500 cm(3) soil. Reniform nematode spread could be explained by the direction of tillage and water flow in the low end of the field. Highest population densities were observed in the areas of the field with soil types ranging from 54% to 60% silt fraction. In addition to R. reniformis, Meloidogyne incognita was commonly detected in many of the grids, and Tylenchorhynchus spp., Helicotylenchus spp., Paratrichodorus minor and Hoplolaimus magnistylus were detected occasionally.


Neural dynamics of motion processing and speed discrimination.
A neural network model of visual motion perception and speed discrimination is presented. The model shows how a distributed population code of speed tuning, that realizes a size-speed correlation, can be derived from the simplest mechanisms whereby activations of multiple spatially short-range filters of different size are transformed into speed-turned cell responses. These mechanisms use transient cell responses to moving stimuli, output thresholds that covary with filter size, and competition. These mechanisms are proposed to occur in the V1-->MT cortical processing stream. The model reproduces empirically derived speed discrimination curves and simulates data showing how visual speed perception and discrimination can be affected by stimulus contrast, duration, dot density and spatial frequency. Model motion mechanisms are analogous to mechanisms that have been used to model 3-D form and figure-ground perception. The model forms the front end of a larger motion processing system that has been used to simulate how global motion capture occurs, and how spatial attention is drawn to moving forms. It provides a computational foundation for an emerging neural theory of 3-D form and motion perception.


Understanding the role and deployment of volunteers within specialist palliative care services and organisations as they have adjusted to the COVID-19 pandemic: A multi-national EAPC volunteer taskforce survey.
Early indications were of a major decline in specialist palliative care volunteer numbers during COVID-19. It is important that ongoing deployment and role of volunteers is understood, given the dependence of many palliative care services on volunteers for quality care provision. To understand the roles and deployment of volunteers in specialist palliative care services as they have adjusted to the impact of COVID-19. Observational multi-national study, using a cross-sectional online survey with closed and free-text option questions. Disseminated via social media, palliative care networks and key collaborators from May to July 2021. Any specialist palliative care setting in any country, including hospices, day hospices, hospital based or community teams. The person responsible for managing the deployment of volunteers was invited to complete the survey. Valid responses were received from 304 organisations (35 countries, 80.3% Europe). Most cared for adults only (60.9%), provided in-patient care (62.2%) and were non-profit (62.5%). 47.0% had cared for people with COVID-19. 47.7% changed the way they deployed volunteers; the mean number of active volunteers dropped from 203 per organisation to 33, and 70.7% reported a decrease in volunteers in direct patient/family facing roles. There was a shift to younger volunteers. 50.6% said this drop impacted care provision, increasing staff workload and pressure, decreasing patient support, and increasing patient isolation and loneliness. The sustained reduction in volunteer deployment has impacted the provision of specialist palliative care. Urgent consideration must be given to the future of volunteering including virtual modes of delivery, micro-volunteering, and appealing to a younger demographic.


Adapting to climate change on Western public lands: addressing the ecological effects of domestic, wild, and feral ungulates.
Climate change affects public land ecosystems and services throughout the American West and these effects are projected to intensify. Even if greenhouse gas emissions are reduced, adaptation strategies for public lands are needed to reduce anthropogenic stressors of terrestrial and aquatic ecosystems and to help native species and ecosystems survive in an altered environment. Historical and contemporary livestock production-the most widespread and long-running commercial use of public lands-can alter vegetation, soils, hydrology, and wildlife species composition and abundances in ways that exacerbate the effects of climate change on these resources. Excess abundance of native ungulates (e.g., deer or elk) and feral horses and burros add to these impacts. Although many of these consequences have been studied for decades, the ongoing and impending effects of ungulates in a changing climate require new management strategies for limiting their threats to the long-term supply of ecosystem services on public lands. Removing or reducing livestock across large areas of public land would alleviate a widely recognized and long-term stressor and make these lands less susceptible to the effects of climate change. Where livestock use continues, or where significant densities of wild or feral ungulates occur, management should carefully document the ecological, social, and economic consequences (both costs and benefits) to better ensure management that minimizes ungulate impacts to plant and animal communities, soils, and water resources. Reestablishing apex predators in large, contiguous areas of public land may help mitigate any adverse ecological effects of wild ungulates.


Molecular identification of blood-meal sources in Culiseta melanura and Culiseta morsitans from an endemic focus of eastern equine encephalitis virus in New York.
Eastern equine encephalitis (EEE) virus perpetuates in an enzootic cycle involving ornithophilic mosquito vectors, principally Culiseta melanura (Coquillett) and avian amplification hosts. To better understand the role of Cs. melanura and Culiseta morsitans (Theobald) in the epizootiology of EEE virus, we collected blood-fed mosquitoes between 31 May and 15 October 2004 at two sites associated with an EEE virus focus in central New York and identified the source of vertebrate blood by nucleotide sequencing of polymerase chain reaction (PCR) products of the cytochrome b gene. Analysis of 484 Cs. melanura and 122 Cs. morsitans revealed that 94.2% and 86.9%, respectively, acquired blood solely from avian hosts. Blood meals derived exclusively from mammals were detected in 0.8% of Cs. melanura and 1.6% of Cs. morsitans. Individual mosquitoes containing mixed-blood meals from both avian and mammalian hosts were also detected in 5.0% of Cs. melanura and 11.5% of Cs. morsitans. Wood thrush constituted the most common vertebrate host for Cs. melanura (23.6%) and Cs. morsitans (30.9%), followed by American robin, song sparrow, ovenbird, red-eyed vireo, and common yellowthroat. Mammalian-derived blood meals were identified as white-tailed deer, horse, domestic cat, and eastern pipistrelle bat. There were three isolations of EEE virus from Cs. melanura and one from Cs. morsitans. These results suggest that wood thrush and a few other passerine birds may play key roles in supporting EEE virus transmission in the northeast and possibly throughout the geographic range of EEE in North America. The frequency of mammalian feedings also suggests that Cs. melanura and Cs. morsitans may play a role in the transmission of EEE virus to equines, in addition to maintaining enzootic transmission among avian hosts. We report the first isolation of arboviruses from mosquito vectors concomitant with the identifications of their blood meal sources.


Pathways accessory to proteasomal proteolysis are less efficient in major histocompatibility complex class I antigen production.
Degradation of cytosolic proteins depends largely on the proteasome, and a fraction of the cleavage products are presented as major histocompatibility complex (MHC) class I-bound ligands at the cell surface of antigen presenting cells. Proteolytic pathways accessory to the proteasome contribute to protein turnover, and their up-regulation may complement the proteasome when proteasomal proteolysis is impaired. Here we show that reduced reliance on proteasomal proteolysis allowed a reduced efficiency of MHC class I ligand production, whereas protein turnover and cellular proliferation were maintained. Using the proteasomal inhibitor adamantane-acetyl-(6-aminohexanoyl)3-(leucinyl)3-vinyl-(methyl)-sulphone, we show that covalent inhibition of all three types of proteasomal beta-subunits (beta(1), beta(2), and beta(5)) was compatible with continued growth in cells that up-regulate accessory proteolytic pathways, which include cytosolic proteases as well as deubiquitinating enzymes. However, under these conditions, we observed poor assembly of H-2D(b) molecules and inhibited presentation of endogenous tumor antigens. Thus, the tight link between protein turnover and production of MHC class I ligands can be broken by enforcing the substitution of the proteasome with alternative proteolytic pathways.


Advances in understanding and treating ADHD.
Attention deficit hyperactivity disorder (ADHD) is a neurocognitive behavioral developmental disorder most commonly seen in childhood and adolescence, which often extends to the adult years. Relative to a decade ago, there has been extensive research into understanding the factors underlying ADHD, leading to far more treatment options available for both adolescents and adults with this disorder. Novel stimulant formulations have made it possible to tailor treatment to the duration of efficacy required by patients, and to help mitigate the potential for abuse, misuse and diversion. Several new non-stimulant options have also emerged in the past few years. Among these, cognitive behavioral interventions have proven popular in the treatment of adult ADHD, especially within the adult population who cannot or will not use medications, along with the many medication-treated patients who continue to show residual disability.


Distribution and biology of Ornithodoros erraticus in parts of Spain affected by African swine fever.
Ornithodoros erraticus was found in 30.7 per cent, 35.0 per cent and 71.0 per cent of the pig-pens sampled in the provinces of Salamanca, Badajoz and Huelva in which African swine fever is a problem in the rearing of Iberian pigs. Between 38 and 65 per cent of the pig-pens in these areas are now abandoned and their populations of O erraticus are extinct or becoming so because they can no longer feed on pigs, which in Spain are their main hosts. The abandonment of pig-pens has resulted in the elimination of most soft ticks infected with the virus of African swine fever, and means that the distribution of ticks is now irregular and focal. Another factor affecting their distribution is the kind of soil on which the pig-pens are located. In abandoned pig-pens, the adults and large nymphs survive for about five years or longer when animals occasionally enter them. Hungry tick populations may transmit African swine fever when feeding in winter, whereas the populations that have continuous access to pigs do not feed until the pig-pens reach a temperature of 13 to 15 degrees C. In the latter populations, each stage exhibits a single annual peak of activity, which implies that the development from larva to adult takes two to three years. Pigs may die as a result of the bites, but on no occasion were 100 per cent of the fasting ticks seen to feed, even though they had the opportunity of doing so. This may hinder the eradication of this soft tick from infested pig-pens.


Early destruction of tumor vasculature in tumor necrosis factor-alpha-based isolated limb perfusion is responsible for tumor response.
Addition of high-dose tumor necrosis factor-alpha to melphalan-based isolated limb perfusion enhances anti-tumor effects impressively. Unfortunately, the mechanism of action of tumor necrosis factor-alpha is still not fully understood. Here, we investigated the effects of tumor necrosis factor-alpha on the tumor microenvironment and on secondary immunological events during and shortly after isolated limb perfusion in soft-tissue sarcoma-bearing rats. Already during isolated limb perfusion, softening of the tumor was observed. Co-administration of tumor necrosis factor-alpha in the isolated limb perfusion with melphalan induced a six-fold enhanced drug accumulation of melphalan in the tumor compared with isolated limb perfusion with melphalan alone. In addition, directly after perfusion with tumor necrosis factor-alpha plus melphalan, over a time-frame of 30 min, vascular destruction, erythrocyte extravasation and hemorrhage was detected. Interstitial fluid pressure and pH in the tumor, however, were not altered by tumor necrosis factor-alpha and no clear immune effects, cellular infiltration or cytokine expression were observed. Taken together, these results indicate that tumor necrosis factor-alpha induces rapid damage to the tumor vascular endothelial lining resulting in augmented drug accumulation. As other important parameters were not changed (e.g. interstitial fluid pressure and pH), we speculate that the tumor vascular changes, and concurrent hemorrhage and drug accumulation are the key explanations for the observed synergistic anti-tumor response.


Influence of bifurcations on forced oscillations in an airway model.
Forced oscillations is a technique to determine respiratory input impedance from small amplitude sinusoidal pressure excursions introduced at the airway opening. Models used to predict respiratory input impedance typically ignore the direct effect of bifurcations on the flow, and treat airway branches as individual straight tubes placed appropriately in parallel and series. The flow within the individual tubes is assumed equivalent to that which would occur in infinitely long tubes. In this study we examined the influence of bifurcations on impedance for conditions of the forced oscillatory technique. We measured input impedance using forced oscillations in straight tubes and in an anatomically-relevant, four generation physical model of a human airway network. The input impedance measured experimentally compared well to that obtained theoretically using model predictions. The predictive scheme was based on appropriate parallel and series combinations of theoretically computed individual tube impedances, which were computed from solutions to oscillatory flow of a compressible gas in an infinitely long rigid tube. The agreement between experimental measurements and predictions indicates that bifurcations play a relatively minor direct role on the flow impedance for conditions of the forced oscillations technique. These results are explained in terms of the small tidal volumes used, whereby the axial distance traveled by a fluid particle during an oscillation cycle is appreciably smaller than branch segment lengths. Accordingly, only a small fraction of fluid particles travel through the bifurcation region, and the remainder experience an environment approaching flow in an infinite straight tube. The relevance of the study to the prediction of impedances in the human lung during forced oscillations is discussed.


Changing a urethral or suprapubic catheter: the patient's perspective.
Understanding the patient's perspective in any area of health care is now recognised as being crucial to good practice, but little is known about patients' views on living with an indwelling urinary catheter. In this study, 36 people living with either a urethral or suprapubic catheter across the UK were interviewed. After thematic analysis, the authors interpreted what they learned. Many issues were raised, including the need to know who was responsible for changing the catheter, the importance of continuity of care, 24-hour access to services, and good hygiene. Patients valued nurses with training and experience, with plenty of time, who took care and who listened to what mattered to the patient. The patient's perspective is important at all levels, not only when designing and planning an integrated continence service, but also in delivering services in the community.


Control of daily transcript oscillations in Drosophila by light and the circadian clock.
The transcriptional circuits of circadian clocks control physiological and behavioral rhythms. Light may affect such overt rhythms in two ways: (1) by entraining the clock circuits and (2) via clock-independent molecular pathways. In this study we examine the relationship between autonomous transcript oscillations and light-driven transcript responses. Transcript profiles of wild-type and arrhythmic mutant Drosophila were recorded both in the presence of an environmental photocycle and in constant darkness. Systematic autonomous oscillations in the 12- to 48-h period range were detectable only in wild-type flies and occurred preferentially at the circadian period length. However, an extensive program of light-driven expression was confirmed in arrhythmic mutant flies. Many light-responsive transcripts are preferentially expressed in the compound eyes and the phospholipase C component of phototransduction, NORPA (no receptor potential), is required for their light-dependent regulation. Although there is evidence for the existence of multiple molecular clock circuits in cyanobacteria, protists, plants, and fungi, Drosophila appears to possess only one such system. The sustained photic expression responses identified here are partially coupled to the circadian clock and may reflect a mechanism for flies to modulate functions such as visual sensitivity and synaptic transmission in response to seasonal changes in photoperiod.