Functional and genetic interactions of TOR in the budding yeast Saccharomyces cerevisiae with myosin type II-deficiency (myo1Δ).
Yeast has numerous mechanisms to survive stress. Deletion of myosin type II (myo1Δ) in Saccharomyces cerevisiae results in a cell that has defective cytokinesis. To survive this genetically induced stress, this budding yeast up regulates the PKC1 cell wall integrity pathway (CWIP). More recently, our work indicated that TOR, another stress signaling pathway, was down regulated in myo1Δ strains. Since negative signaling by TOR is known to regulate PKC1, our objectives in this study were to understand the cross-talk between the TOR and PKC1 signaling pathways and to determine if they share upstream regulators for mounting the stress response in myo1Δ strains. Here we proved that TORC1 signaling was down regulated in the myo1Δ strain. While a tor1Δ mutant strain had increased viability relative to myo1Δ, a combined myo1Δtor1Δ mutant strain showed significantly reduced cell viability. Synthetic rescue of the tor2-21(ts) lethal phenotype was observed in the myo1Δ strain in contrast to the chs2Δ strain, a chitin synthase II null mutant that also activates the PKC1 CWIP and exhibits cytokinesis defects very similar to myo1Δ, where the rescue effect was not observed. We observed two pools of Slt2p, the final Mitogen Activated Protein Kinase (MAPK) of the PKC1 CWIP; one pool that is up regulated by heat shock and one that is up regulated by the myo1Δ stress. The cell wall stress sensor WSC1 that activates PKC1 CWIP under other stress conditions was shown to act as a negative regulator of TORC1 in the myo1Δ mutant. Finally, the repression of TORC1 was inversely correlated with the activation of PKC1 in the myo1Δ strain. Regulated expression of TOR1 was important in the activation of the PKC1 CWIP in a myo1Δ strain and hence its survival. We found evidence that the PKC1 and TORC1 pathways share a common upstream regulator associated with the cell wall stress sensor WSC1. Surprisingly, essential TORC2 functions were not required in the myo1Δ strain. By understanding how yeast mounts a concerted stress response, one can further design pharmacological cocktails to undermine their ability to adapt and to survive.


The illusion of increasing loudness in brief steady tones: variation with carrier frequency.
A brief tone of steady intensity is heard as growing louder; to be heard as steady, intensity must be decreasing. The present report concerns the influence of carrier frequency on this illusion. Stimuli each lasted 1.5 s, during which time intensity was increasing, decreasing, or remaining steady; the initial intensity was 40 dB SPL (sound pressure level relative to 0.0002 dynes/cm2). Carrier frequencies were between 0.0625 and 8.0 kHz. Four listeners made forced binary responses of "growing louder" or "growing softer" to stimuli. Values of changing intensity that elicited equal numbers of each type of response were computed. As expected, these values were negative. The illusion was most pronounced for the lowest and highest frequencies. In contrast to the results of a recent study, the findings were that sensitivity to changing intensity did not vary systematically with the size of the illusion. The illusion might arise because many sounds slowly decay in intensity and because of the importance of detecting approaching sound sources, analogous to "looming" in the visual modality. Overall loudness of the sound source should assist in altering the listener; the increase in the illusion at the extremities of the frequency range might compensate for the reduced overall loudness at such frequencies.


Multicentre research and the WISECARE experience. Workflow Information Systems for European Nursing Care.
The benefits of collaborative research are becoming recognized increasingly within the profession of nursing due to the associated increased likelihood of funding, variety of practice settings and increased access to resources that collaboration brings. While such benefits have made collaborative research one of the most desirable strategies for achieving the goals of research, the potential problems of language and communication, culturally sensitive instruments, access to subjects, availability of technology and lack of research resources have complicated collaborative international research initiatives. Review of the literature, although encouraging such initiatives, does little to provide information regarding the processes involved in multinational collaboration or the associated advantages and disadvantages to guide those embarking on such large scale, multinational, cross-cultural studies. The diverse meanings of collaboration within research initiatives further hamper this understanding. Positive definitions focus on aspects such as sharing expertise, making a valuable contribution to the research and ultimately shared ownership of the accomplishments of the research. One such research project led by nurses was the WISECARE project. WISECARE (Workflow Information Systems for European Nursing Care) was funded by the European Commission and aimed to improve cancer nursing practice and ultimately patient outcomes through the integration and utilization of state of the art information technology. Such a project was developed as a result of nursing's apparent invisibility within health care delivery and the problems experienced by nurses in articulating their worth within an increasingly cost-conscious health care system. Oncology care was selected as the domain for the project not only because this speciality of nursing already has an established network of nurses throughout Europe in the European Oncology Nursing Society (EONS) but also because the practice of cancer nursing encompasses all aspects of nursing care. This paper will address the advantages and disadvantages associated with collaboration, using the WISECARE project as an example.


Epigenomics: sequencing the methylome.
DNA methylation patterns are increasingly surveyed through methods that utilize massively parallel sequencing. Sequence-based assays developed to detect DNA methylation can be broadly divided into those that depend on affinity enrichment, chemical conversion, or enzymatic restriction. The DNA fragments resulting from these methods are uniformly subjected to library construction and massively parallel sequencing. The sequence reads are subsequently aligned to a reference genome and subjected to specialized analytical tools to extract the underlying methylation signature. This chapter will outline these emerging techniques.


Complex consultations in primary care: a tool for assessing the range of health problems and issues addressed in general practice consultations.
There is an increasing recognition that many consultations in general practice involve several problems covering multiple disease domains. However there is a paucity of reliable tools and techniques to understand and quantify this phenomenon. The objective was to develop a tool that can be used to measure the number and type of problems discussed in primary care consultations. Thirteen consultations between general practitioners and patients were initially videoed and reviewed to identify the problems and issues discussed. An iterative process involving a panel of clinicians and researchers and repeated cycles of testing and development was used to develop a measurement proforma and coding manual for assessment of video recorded consultations. The inter-rater reliability of this tool was assessed in 60 consultations. The problems requiring action were usually readily identified. However the different dimensions of the problem and how they were addressed required the identification and definition of 'issues'. A coding proforma was developed that allowed quantification of the numbers and types of health problems and issues discussed. Ten categories of issues were identified and defined. At the consultation level, inter-rater agreements for the number of problems discussed (within ± 1), types of problems and issues were 98.3%, 96.5% and 90% respectively. The tool has subsequently been used to analyse 229 consultations. The iterative approach to development of the tool reflected the complexity of doctor-patient interactions. A reliable tool has been developed that can be used to analyse the number and range of problems managed in primary care consultations.


Complete cytogenetic and molecular responses to interferon-alpha-based therapy for chronic myelogenous leukemia are associated with excellent long-term prognosis.
Little is known regarding long-term prognosis among patients with Philadelphia chromosome (Ph)-positive chronic myelogenous leukemia (CML) who achieve a complete cytogenetic response (0% Ph-positive cells) after treatment with interferon-alpha. The authors analyzed 512 patients with Ph-positive, early chronic-phase CML who were treated with interferon-based therapies between 1981-1995 for the incidence and durability of complete cytogenetic response, and in relation to long-term prognosis. One hundred forty patients (27%) achieved a complete cytogenetic response. Their 10-year survival rate was 78%. At the time of last follow-up, 44 patients (31%, 9% of the total) were alive, 21 in first and 23 in second durable complete cytogenetic response (median, 127 months; range, 88-191 months); 39 patients had not received any therapy for a median of 50 months (range, 11-139 months). Analysis by reverse transcriptase-polymerase chain reaction in 78 patients during complete cytogenetic response showed 46 who had achieved at least 1 complete molecular response. Five of these 78 patients had died by the time of last follow-up, but only 2 had died of disease-specific causes. Recurrence rates were significantly lower and cytogenetic response durations were significantly longer among patients who achieved at least one complete molecular response. Achieving a complete cytogenetic or molecular response after therapy with interferon-alpha appears to be associated with excellent long-term prognosis. Approximately 10% of patients reportedly can achieve durable complete cytogenetic response, with or without continuation of interferon. This finding emphasizes the potential of long-term event-free survival in CML patients outside the context of allogeneic stem cell transplantation, which may be improved with new therapies such as imatinib mesylate.


Stabilization of a Virus-Like Particle and Its Application as a Nanoreactor at Physiological Conditions.
Virus-like particles are very interesting tools for application in bionanotechnology, due to their monodisperse features and biocompatibility. In particular, the cowpea chlorotic mottle virus (CCMV) capsid has been studied extensively as it can be assembled and disassembled reversibly, facilitating cargo encapsulation. CCMV is, however, only stable at physiological conditions when its endogenous nucleic acid cargo is present. To gain more flexibility in the type of cargo encapsulated and to broaden the window of operation, it is interesting to improve the stability of the empty virus-like particles. Here, a method is described to utilize the CCMV capsid at close to physiological conditions as a stable, enzyme-filled nanoreactor. As a proof-of-principle, the encapsulation of T4 lysozyme (T4L) was chosen; this enzyme is a promising antibiotic, but its clinical application is hampered by, for example, its cationic character. It was shown that four T4L molecules can successfully be encapsulated inside CCMV capsids, while remaining catalytically active, which could thus improve the enzyme's application potential.


Humidity adsorption kinetics of a trypsin gel film.
This study focuses on the humidity adsorption kinetics of an isopropanol-induced and pH-triggered bovine pancreatic trypsin gel (BPTG). The BPTG was adsorbed on a gold coated Quartz Crystal Microbalance (QCM) substrate with a thickness of 376 nm. The morphology of the film was characterized using Atomic Force Microscopy (AFM). QCM was used to investigate the humidity sensing properties of the BPTG film. The response of the humidity sensor was explained using the Langmuir model. The average values of adsorption and desorption rates between 11% RH (relative humidity) and 97% RH were calculated as 2482.5 M(-1) s(-1) and 0.02 s(-1), respectively. The equilibrium constant and average Gibbs Free Energy of humidity adsorption and desorption cycles were obtained as 133,000 and -11.8 kJ/mol, respectively.


Estimation of Lipid Hydroperoxide Levels in Tropical Marine Macroalgae.
The incipient levels of lipid hydroperoxides (LHPOs) were determined in selected green, brown, and red macroalgae by the FOX assay using hydroperoxy HPLC mix. The LHPOs contents varied between the investigated species and showed relatively low values in this study. Among the greens, it varied from 12 ± 6.2 μg · g(-1) (Codium sursum) to 31.5 ± 2.8 μg · g(-1) (Ulva lactuca), whereas in reds, from 5.7 ± 1.6 μg · g(-1) (Gracilaria corticata) to 46.2 ± 6 μg · g(-1) (Sarconema filiforme), and in browns, from 4.6 ± 4.4 μg · g(-1) (Dictyota bartayresiana) to 79 ± 5.0 μg · g(-1) (Sargassum tenerrimum), on fresh weight basis. These hydroperoxides represented a minor fraction of total lipids and ranged from 0.04% (S. swartzii) to 1.1% (S. tenerrimum) despite being a rich source of highly unsaturated fatty acids. The susceptibility of peroxidation was assessed by specific lipid peroxidazibility (SLP) values for macroalgal tissues. The LHPO values were found to be independent of both the PUFAs contents and their degree of unsaturation (DBI), as evident from the PCA analysis. SLP values were positively correlated with the LHPOs and negatively with DBI. The FOX assay gave ≥20-fold higher values for LHPOs as compared to the TBARS method for all the samples investigated in this study. Furthermore, U. lactuca cultured in artificial seawater (ASW) enriched with nutrients (N, P, and NP) showed a sharp decline in LHPOs contents relative to those cultured in ASW alone P ≤ 0.05. It is inferred from this study that the FOX assay is an efficient, rapid, sensitive, and inexpensive technique for detecting the incipient lipid peroxidation in macroalgal tissues.


Functional analysis of killer Ig-like receptor-expressing cytomegalovirus-specific CD8+ T cells.
Killer Ig-like receptors (KIR) are expressed by human NK cells and T cells. Although Ag-specific cytolytic activity and cytokine production of KIR(+) T cells can be inhibited by KIR ligation, the effect of KIR on proliferation is unclear. KIR(+) T cells have been reported to have a general proliferative defect. To investigate whether KIR(+) T cells represent end-stage dysfunctional T cells, we characterized KIR(+) CMV-specific T cells in allogeneic stem cell transplantation patients and healthy donors. In both patients and healthy donors, a significant percentage KIR(+) T cells was detected at various time points. All stem cell transplantation patients studied showed KIR expression on CMV-specific T cells, while not all donors had KIR-expressing CMV-specific T cells. From two of the patients and one donor KIR(+) CMV-specific T clones were isolated and analyzed functionally. T cells were detected that expressed KIR that could not encounter their corresponding KIR ligands in vivo, illustrating that KIR expression by these T cells was not based on functional selection but a random process. Our data demonstrate that KIR(+) T cells are fully functional T cells that are only restricted in effector functions and proliferation upon KIR ligation. The level of KIR-mediated inhibition of the effector functions and proliferation depended on the strength of TCR stimulation. We observed no diminished general proliferative capacity and therefore we conclude that these T cells do not represent end-stage dysfunctional T cells.


Pain in Patients with Dystrophic Epidermolysis Bullosa: Association with Anxiety and Depression.
We investigate the presence and the quality of pain in patients with dystrophic epidermolysis bullosa (DEB), and its correlation with the level of anxiety and depression. We collected data from 27 DEB patients and 26 healthy individuals. DEB patients and controls completed 1 scale for the quality of pain, and 1 scale for anxiety and depression. Pain was assessed with the short form of the McGill Pain Questionnaire, whereas anxiety and depression were assessed with the Hamilton rating scale for anxiety and depression. DEB patients and healthy control individuals were homogeneous for age and gender (p>0.05). A statistically significant difference in the two groups was seen for sensory pain rating scale (p<0.001), affective pain rating scale (p=0.029), total pain rating scale (p<0.001), visual analogue scale (p=0.012) and present pain intensity (p=0.001), but not for anxiety (p=0.169) and depression (p=0.530). The characteristics of pain that showed a significant difference between DEB patients and healthy controls were shooting, splitting, tender and throbbing (p<0.05). In DEB patients pain was not correlated with anxiety or depression (p>0.05), whereas a slight correlation between pain and anxiety was found in healthy controls (p<0.05). No difference was found between quality of pain and anxiety-depression in DEB patients (p>0.05), but was between the DEB dominant and the recessive form of DEB (p=0.025). The perception of pain in DEB patients appears greater than in healthy individuals, with splitting and tender characteristics being the most significant ones, but was not associated with anxious and/or depressive symptoms.


Miscoding, misclassification and misdiagnosis of diabetes in primary care.
To determine the effectiveness of self-audit tools designed to detect miscoding, misclassification and misdiagnosis of diabetes in primary care. We developed six searches to identify people with diabetes with potential classification errors. The search results were automatically ranked from most to least likely to have an underlying problem. Eight practices with a combined population of 72,000 and diabetes prevalence 2.9% (n = 2340) completed audit forms to verify whether additional information within the patients' medical record confirmed or refuted the problems identified. The searches identified 347 records, mean 42 per practice. Pre-audit 20% (n = 69) had Type 1 diabetes, 70% (n = 241) had Type 2 diabetes, 9% (n = 30) had vague codes that were hard to classify, 2% (n = 6) were not coded and one person was labelled as having gestational diabetes. Of records, 39.2% (n = 136) had important errors: 10% (n = 35) had coding errors; 12.1% (42) were misclassified; and 17.0% (59) misdiagnosed as having diabetes. Thirty-two per cent (n = 22) of people with Type 2 diabetes (n = 69) were misclassified as having Type 1 diabetes; 20% (n = 48) of people with Type 2 diabetes (n = 241) did not have diabetes; of the 30 patients with vague diagnostic terms, 50% had Type 2 diabetes, 20% had Type 1 diabetes and 20% did not have diabetes. Examples of misdiagnosis were found in all practices, misclassification in seven and miscoding in six. Volunteer practices successfully used these self-audit tools. Approximately 40% of patients identified by computer searches (5.8% of people with diabetes) had errors; misdiagnosis is commonest, misclassification may affect treatment options and miscoding in omission from disease registers and the potential for reduced quality of care.


Vestibular Rehabilitation After Traumatic Brain Injury: Case Series.
There has been an increasing focus on vestibular rehabilitation (VR) after traumatic brain injury (TBI) in recent years. However, detailed descriptions of the content of and patient responses to VR after TBI are limited. The purposes of this case series are (1) to describe a modified, group-based VR intervention and (2) to examine changes in self-reported and performance-based outcome measures. Two women and 2 men (aged 24-45 years) with mild TBI, dizziness, and balance problems participated in an 8-week intervention consisting of group sessions with guidance, individually modified VR exercises, a home exercise program, and an exercise diary. Self-reported and performance-based outcome measures were applied to assess the impact of dizziness and balance problems on functions related to activity and participation. The intervention caused no adverse effects. Three of the 4 patients reported reduced self-perceived disability because of dizziness, diminished frequency and severity of dizziness, improved health-related quality of life, reduced psychological distress, and improved performance-based balance. The change scores exceeded the minimal detectable change, indicating a clinically significant change or improvement in the direction of age-related norms. The fourth patient did not change or improve in most outcome measures. A modified, group-based VR intervention was safe and appeared to be viable and beneficial when addressing dizziness and balance problems after TBI. However, concurrent physical and psychological symptoms, other neurological deficits, and musculoskeletal problems might influence the course of central nervous system compensation and recovery. The present case series may be useful for tailoring VR interventions to patients with TBI. Future randomized controlled trials are warranted to evaluate the short- and long-term effects of VR after TBI.


Suspected Pediatric Influenza Risk-Stratification Algorithm: A Clinical Decision Tool.
Influenza causes significant annual burden among children. Current guidelines recommend empiric treatment for a broadly defined group of children at high risk for influenza complications, resulting in overtreatment or costly viral testing. This study creates an algorithm for clinicians to risk stratify children with influenza-like illness (ILI) according to likelihood of influenza infection. A retrospective analysis was performed on 818 children seen in the emergency department from November 2012 to April 2013 for ILI. We reviewed medical records for symptoms, influenza risk factors, and viral assay results. Classification and regression tree analyses were performed separately for children older and younger than 2 years. In children younger than 2 years, populations likely to test positive were those with an influenza-positive contact, unimmunized children, and those presenting in high-incidence influenza periods. In this subgroup, immunized patients in low-incidence seasons and those with absence of cough are low risk for influenza infection. For children 2 years and older, high-risk populations were unimmunized children, those presenting in high-incidence influenza periods and those with myalgia or absence of diarrhea. These risk-stratification analyses were summarized into Suspected Pediatric Influenza Risk-Stratification Algorithm (SPIRA). For those in whom influenza infection is likely, clinicians may consider empiric treatment. Conversely, patients whom SPIRA identifies as unlikely to be infected with influenza are candidates for viral testing and targeted treatment. In assessing children with ILI, SPIRA aids clinicians in determining who to test versus treat empirically, saving children from costly viral testing or unnecessary antiviral exposure.


Estimating physical activity energy expenditure, sedentary time, and physical activity intensity by self-report in adults.
Few questionnaires that assess usual physical activity have been reported to be valid for all different subdimensions of physical activity. The objective was to assess the validity and reliability of the Recent Physical Activity Questionnaire (RPAQ), which assesses usual physical activity (PA) in 4 domains (work, travel, recreation, and domestic life). Total energy expenditure (TEE) was measured for 14 d by using the doubly labeled water technique combined with a measure of resting metabolic rate to yield PA energy expenditure (PAEE) in 25 men and 25 women. Simultaneously, intensity of activity was measured by using combined heart rate and movement sensing for 11 d. Repeatability of the RPAQ was assessed in an independent sample of 71 women and 60 men aged 31-57 y. Estimated TEE and PAEE were significantly associated with criterion measures (TEE: r = 0.67; PAEE: r = 0.39) with mean (+/-SD) biases of -3452 +/- 2025 kJ/d and -13 +/- 24 kJ d(-1) kg(-1). The correlation between self-reported and measured time spent was significant for vigorous PA (r = 0.70) and marginally insignificant for sedentary time (r = 0.27, P = 0.06). The mean biases were relatively small for sedentary time and vigorous PA: 0.7 +/- 2.8 h/d and -12 +/- 24 min/d, respectively. The intraclass correlation coefficient for repeatability of total PAEE (kJ/d) was 0.76 (P < 0.0001). The RPAQ is the first questionnaire with demonstrated validity for ranking individuals according to their time spent at vigorous-intensity activity and overall energy expenditure.


Regulation of lymphocyte development and function by RNA-binding proteins.
Lymphocyte development requires cells to progress through a series of stages, each associated with changes in gene expression. Intense effort has been invested into characterising the dynamic networks of transcription factors underlying these regulated changes. Whilst transcription factors determine the tempo at which mRNA is produced, recent results highlight the importance of the selective regulation of mRNA decay and translation in regulating gene expression. These processes are regulated by sequence-specific RNA-binding proteins (RBP) as well as noncoding RNA such as microRNAs. RNA-binding proteins are emerging as important regulators of cell fate and function in both developing and mature lymphocytes. At the molecular level the function of RNA-binding proteins is integrated with signal transduction pathways that also govern gene transcription.


Cardiovascular complications of radiation therapy for thoracic malignancies: the role for non-invasive imaging for detection of cardiovascular disease.
Radiation exposure to the thorax is associated with substantial risk for the subsequent development of cardiovascular disease. Thus, the increasing role of radiation therapy in the contemporary treatment of cancer, combined with improving survival rates of patients undergoing this therapy, contributes to a growing population at risk of cardiovascular morbidity and mortality. Associated cardiovascular injuries include pericardial disease, coronary artery disease, valvular disease, conduction disease, cardiomyopathy, and medium and large vessel vasculopathy-any of which can occur at varying intervals following irradiation. Higher radiation doses, younger age at the time of irradiation, longer intervals from the time of radiation, and coexisting cardiovascular risk factors all predispose to these injuries. The true incidence of radiation-related cardiovascular disease remains uncertain due to lack of large multicentre studies with a sufficient duration of cardiovascular follow-up. There are currently no consensus guidelines available to inform the optimal approach to cardiovascular surveillance of recipients of thoracic radiation. Therefore, we review the cardiovascular consequences of radiation therapy and focus on the potential role of non-invasive cardiovascular imaging in the assessment and management of radiation-related cardiovascular disease. In doing so, we highlight characteristics that can be used to identify individuals at risk for developing post-radiation cardiovascular disease and propose an imaging-based algorithm for their clinical surveillance.


An Event Related Field Study of Rapid Grammatical Plasticity in Adult Second-Language Learners.
The present study used magnetoencephalography (MEG) to investigate how Spanish adult learners of Basque respond to morphosyntactic violations after a short period of training on a small fragment of Basque grammar. Participants (n = 17) were exposed to violation and control phrases in three phases (pretest, training, generalization-test). In each phase participants listened to short Basque phrases and they judged whether they were correct or incorrect. During the pre-test and generalization-test, participants did not receive any feedback. During the training blocks feedback was provided after each response. We also ran two Spanish control blocks before and after training. We analyzed the event-related magnetic- field (ERF) recorded in response to a critical word during all three phases. In the pretest, classification was below chance and we found no electrophysiological differences between violation and control stimuli. Then participants were explicitly taught a Basque grammar rule. From the first training block participants were able to correctly classify control and violation stimuli and an evoked violation response was present. Although the timing of the electrophysiological responses matched participants' L1 effect, the effect size was smaller for L2 and the topographical distribution differed from the L1. While the L1 effect was bilaterally distributed on the auditory sensors, the L2 effect was present at right frontal sensors. During training blocks two and three, the violation-control effect size increased and the topography evolved to a more L1-like pattern. Moreover, this pattern was maintained in the generalization test. We conclude that rapid changes in neuronal responses can be observed in adult learners of a simple morphosyntactic rule, and that native-like responses can be achieved at least in small fragments of second language.


Effect of particle size and deformation behaviour on water ingress into tablets.
Drug release performance of tablets is often highly dependent on disintegration, and water ingress is typically the rate-limiting step of the disintegration process. Water ingress into tablets is known to be highly influenced by the microstructure of the tablet, particularly tablet porosity. Initial particle size distribution of the formulation and the predominant powder deformation behaviour during compression are expected to impact such microstructure, making both factors important to investigate in relation to water ingress into tablets. Two size fractions (<125 and 355-500 µm) of plastically deforming microcrystalline cellulose (MCC) and fragmenting di-calcium phosphate (DCP) were compressed into tablets with porosities ranging from 5 to 30% (with 5% increments). The total porosity of the tablets was measured using terahertz time-domain spectroscopy and liquid transport into these tablets was quantified using a flow cell coupled to terahertz pulsed imaging. It was found that tablets compressed from large MCC particles resulted in slower water ingress compared to tablets prepared from small MCC particles. In contrast, no difference in liquid transport kinetics was observed for tablets prepared across both size fractions of DCP particles. These results highlight the complex interplay between material characteristics, the process induced microstructure, and the liquid transport process that ultimately determines the drug release performance of the tablets.


Calpain inhibition but not reticulum endoplasmic stress preconditioning protects rat kidneys from p-aminophenol toxicity.
p-Aminophenol (pAP, 225 mg/kg) administration to rats induced renal failure and has been associated with markers of endoplasmic reticulum (ER) stress, as well as calpain and caspase-12 activation in kidneys. To determine the importance of ER stress and calpain during pAP-induced nephrotoxicity, rats were pretreated with low, nontoxic, doses of ER stress inducers or with the selective calpain inhibitor PD150606 (3 mg/kg). Prior ER stress induced by tunicamycin and oxidized dithiothreitol did not result in protection against renal failure, but PD150606 administration was protective and decreased significantly the rise in creatinine and blood urea nitrogen observed after 24-h post-pAP administration. pAP-induced XBP1 upregulation was not modified by calpain inhibition, but a trend to lower GRP94 induction was determined, suggesting that pAP-induced ER stress was mostly calpain independent. In contrast, pAP-induced caspase-12 cleavage products were significantly decreased with PD150606 pretreatment, demonstrating that caspase-12 activation was calpain dependent. This study reveals the importance of calpain in pAP-induced renal failure. Further research with other nephrotoxicants needs to be performed to determine if calpain activation is a common feature of drug-induced renal failure.


HIV-negative gay men's perceived HIV risk hierarchy: imaginary or real?
HIV-related risk perceptions and risk practices among gay men have changed over time. We revisited perceived HIV risk and engagement in anal intercourse with casual partners among HIV-negative gay men who participated in one of the Sydney Gay Community Periodic Surveys (GCPS). Perceived HIV risk was assessed by a range of anal intercourse practices combined with pre-specified casual partners' HIV status and viral load levels. Perceived HIV risk forms a potential hierarchy, broadly reflecting differences in the probability of HIV transmission through various anal intercourse practices. To a lesser extent, it also varies by casual partners' HIV status and viral load. Men who had unprotected anal intercourse with casual partners (UAIC) perceived lower HIV risk than those who used condoms consistently in the 6 months prior to survey. Recognising the complex associations between risk perceptions and risk practices helps to better address challenges arising from the 'Treatment as Prevention' (TasP).


Dose-dependent Smad1, Smad5 and Smad8 signaling in the early mouse embryo.
Three closely related mammalian R-Smads, namely Smad1, Smad5 and Smad8, are activated by BMP receptors. Here we have taken a genetic approach to further dissect their possibly unique and/or shared roles during early mouse development. A Smad8.LacZ reporter allele was created to visualize Smad8 expression domains. Smad8 is initially expressed only in the visceral yolk sac (VYS) endoderm and shows a highly restricted pattern of expression in the embryo proper at later stages. In addition, Smad8 conditional and null alleles were engineered. All alleles clearly demonstrate that adult Smad8 homozygous mutants are viable and fertile. To elucidate gene dosage effects, we manipulated expression ratios of the three BMP R-Smads. Smad8 homozygotes also lacking one copy of Smad1 or Smad5 did not exhibit overt phenotypes, and the tissue disturbances seen in Smad1 or Smad5 null embryos were not exacerbated in the absence of Smad8. However, we discovered a profound genetic interaction between Smad1 and Smad5. Thus, as for Smad1 and Smad5 mutant embryos, Smad1+/-:Smad5+/- double heterozygotes die by E10.5 and display defects in allantois morphogenesis, cardiac looping and primordial germ cell (PGC) specification. These experiments demonstrate for the first time that Smad1 and Smad5 function cooperatively to govern BMP target gene expression in the early mammalian embryo.


Are brief or recurrent transient global amnesias of epileptic origin?
To evaluate if short (less than one hour) or recurrent, or both, episodes of transient global amnesia (TGA) have an epileptic origin or carry a subsequent risk of epilepsy a group of patients with these types of TGA attacks was studied. The group was selected from a prospective series of 103 patients with TGA. Sixteen patients had an episode lasting less than one hour, 13 had more than one episode, and five patients had both short and recurrent attacks. For each patient the number of recurrences was small (four or less) and they were separated by months or years. During short attacks of TGA many subjects showed other typical features of TGA including repeated questioning (12 subjects) and performance of purposeful complex acts (eight subjects). Twelve short attacks were closely related to a characteristic precipitating event. During follow up only one patient had a seizure (partial motor). No other association between either short or repeated attacks of TGA and past history of epilepsy or paroxysmal discharges were seen on the EEG. Short or recurrent, or both, attacks of TGA are not epileptic and do not carry a relevant risk of subsequent seizures.


Muscarinic Receptor-Dependent Long Term Depression in the Perirhinal Cortex and Recognition Memory are Impaired in the rTg4510 Mouse Model of Tauopathy.
Neurodegenerative diseases affecting cognitive dysfunction, such as Alzheimer's disease and fronto-temporal dementia, are often associated impairments in the visual recognition memory system. Recent evidence suggests that synaptic plasticity, in particular long term depression (LTD), in the perirhinal cortex (PRh) is a critical cellular mechanism underlying recognition memory. In this study, we have examined novel object recognition and PRh LTD in rTg4510 mice, which transgenically overexpress tauP301L. We found that 8-9 month old rTg4510 mice had significant deficits in long- but not short-term novel object recognition memory. Furthermore, we also established that PRh slices prepared from rTg4510 mice, unlike those prepared from wildtype littermates, could not support a muscarinic acetylcholine receptor-dependent form of LTD, induced by a 5 Hz stimulation protocol. In contrast, bath application of the muscarinic agonist carbachol induced a form of chemical LTD in both WT and rTg4510 slices. Finally, when rTg4510 slices were preincubated with the acetylcholinesterase inhibitor donepezil, the 5 Hz stimulation protocol was capable of inducing significant levels of LTD. These data suggest that dysfunctional cholinergic innervation of the PRh of rTg4510 mice, results in deficits in synaptic LTD which may contribute to aberrant recognition memory in this rodent model of tauopathy.


Update on Data Reuse in Health Care.
Objective: Reuse of clinical data has broad use in clinical, research, governmental, and business settings. This summary provides an update on the benefits, barriers to use with large clinical databases, policy frameworks that have been formulated, and challenges. Methods: This report highlights some recent publications on the diverse uses of clinical data and some policy initiatives to promote reuse. It also contains the opinions of the author. Results: Although many examples of the benefits of data reuse have been documented, this summary also reviews why the quality of clinical data needs to be the focus of future informatics work. Conclusion: The promise of reusing data outweighs potential risks, but concerns about privacy and the need to modernize our legal framework will be necessary to realize the full benefits of real-world evidence.


Degradation kinetic study of ZIF-8 microcrystals with and without the presence of lactic acid.
The zeolitic imidazolate framework ZIF-8 (Zn(mim)2, mim = 2-methylimidazolate) has recently been proposed as a drug delivery platform for anticancer therapy based on its capability of decomposing in acidic media. The concept presumes a targeted release of encapsulated drug molecules in the vicinity of tumor tissues that typically produce secretions with elevated acidity. Due to challenges of in vivo and in vitro examination, many studies have addressed the kinetics of ZIF-8 decomposition and subsequent drug release in phosphate buffered saline (PBS) with adjusted acidity. However, the presence of hydrogen phosphate anions [HPO4]2- in PBS may also affect the stability of ZIF-8. As yet, no separate analysis has been performed comparing the dissolving capabilities of PBS and various acidification agents used for regulating pH. Here, we provide a systematic study addressing the effects of phosphate anions with and without lactic acid on the degradation rate of ZIF-8 microcrystals. Lactic acid has been chosen as an experimental acidification agent, since it is particularly secreted by tumor cells. Interestingly, the effect of a lactic acid solution with pH 5.0 on ZIF-8 degradation is shown to be weaker compared to a PBS solution with pH 7.4. However, as an additive, lactic acid is able to enhance the decomposition efficacy of other solutions by 10 to 40 percent at the initial stage, depending on the presence of other ions. Additionally, we report mild toxicity of ZIF-8 and its decomposition products, as examined on HDF and A549 cell lines.


Advancing Diagnostics and Therapy to Reach Universal Cure in Childhood ALL.
Systemic combination chemotherapy and intrathecal chemotherapy markedly increased the survival rate of children with ALL. In the past two decades, the use of minimal (measurable) residual disease (MRD) measurements early in therapy improved risk group stratification with subsequent treatment intensifications for patients at high risk of relapse, and enabled a reduction of treatment for low-risk patients. The recent development of more sensitive MRD technologies may further affect risk stratification. Molecular genetic profiling has led to the discovery of many new subtypes and their driver genetic alterations. This increased our understanding of the biological basis of ALL, improved risk classification, and enabled implementation of precision medicine. In the past decade, immunotherapies, including bispecific antibodies, antibody-drug conjugates, and cellular therapies directed against surface proteins, led to more effective and less toxic therapies, replacing intensive chemotherapy courses and allogeneic stem-cell transplantation in patients with relapsed and refractory ALL, and are now being tested in newly diagnosed patients. It has taken 50-60 years to increase the cure rate in childhood ALL from 0% to 90% by stepwise improvements in chemotherapy. This review provides an overview of how the developments over the past 10-15 years mentioned above have significantly changed the diagnostic and treatment approach in ALL, and discusses how the integrated use of molecular and immunotherapeutic insights will very likely direct efforts to cure those children with ALL who are not cured today, and improve the quality of life for survivors who should have decades of life ahead. Future efforts must focus on making effective, yet very expensive, new technologies and therapies available to children with ALL worldwide.


Regulation of mouse podocyte process dynamics by protein tyrosine phosphatases rapid communication.
Effacement of podocyte foot processes occurs early in many glomerular diseases associated with proteinuria and is accompanied by a reorganization of the actin cytoskeleton. The molecular mechanisms regulating these structural changes are poorly understood. To address these questions, we analyzed the effect of the polycation, protamine sulfate (PS), and puromycin aminonucleoside (PA) on the morphology, cytoskeleton, and tyrosine phosphorylation of differentiated process-bearing cultured podocytes. PS and PA induced similar profound morphological alterations, including retraction and detachment of podocyte processes from the extracellular matrix (ECM). The effects of PS occurred within six hours, whereas PA showed its most severe effects after 72 hours. Structural changes included reorganization of the actin cytoskeleton and focal contacts and were accompanied by an increase in tyrosine phosphorylation. The same effects were induced by application of vanadate, an inhibitor of protein tyrosine phosphatases (PTPs), suggesting that PTPs regulate podocyte process structure. Since disruption of the actin cytoskeleton with cytochalasin B protected the cells from PS-induced effacement and detachment, cytoplasmic PTPs were implicated in these events. Using reverse transcription-polymerase chain reaction (RT-PCR), we demonstrated the expression of four cytoplasmic PTPs in podocytes: SHP-2, PTP-PEST, PTP-1B, and PTP-36. These studies indicate an important role for cytoplasmic PTPs as regulators of podocyte process dynamics. Future studies will aim at restoring the normal foot process architecture of podocytes in glomerular diseases associated with proteinuria by modulating the activity of cytoplasmic PTPs.


Glycoinositol phospholipid anchor and protein C-terminus of bovine erythrocyte acetylcholinesterase: analysis by mass spectrometry and by protein and DNA sequencing.
Purified bovine erythrocyte acetylcholinesterase (AChE) was radiomethylated on its amine groups and incubated with bacterial phosphatidylinositol-specific phospholipase C to remove the lipid portion of the AChE glycoinositol phospholipid (GPI) anchor, and a C-terminal tryptic fragment that contained the residual GPI glycan was isolated by HPLC. Analysis by electrospray-ionization mass spectrometry revealed a parent ion of m/z 3798. The fragmentation patterns produced by collision-induced dissociation mass spectrometry of the +4 and +5 states of the parent ion indicated a 23-amino acid peptide in amide linkage to ethanolamine-P04-Hex-Hex-Hex(PO4-ethanolamine)(HexNAc)-Hex N(Me)2-inositol phosphate. The glycan structure is completely consistent with that obtained previously for the GPI anchor of human erythrocyte AChE except for the addition of the HexNAc substituent. A nearly complete peptide sequence was deduced from the fragmentation patterns, although four assignments were based only on single fragments of very low abundance. To resolve this uncertainty, a segment of bovine genomic DNA corresponding to the C-terminal AChE sequence was amplified by PCR. DNA sequencing established the 23-amino acid peptide sequence to be FLPKLLSATASEAPCTCSGPAHG, in agreement with the MS data and consistent with results from Edman protein sequencing. Dimerization of AChE polypeptides is mediated by intersubunit disulphide bonding in this C-terminal segment, but the bovine AChE contained two cysteine residues in a ...CTC... motif, in contrast with human AChE which contains only a single cysteine in this segment. Although bovine AChE contained no free thiol groups reactive with iodo[14C]acetamide, partial reduction and alkylation with iodo[14C]acetamide revealed that conversion into monomers occurred with an overall incorporation of only one alkyl group per monomer. An identical level of alkylation was observed when dimeric human AChE was converted into monomers by partial reduction. The question of whether the bovine AChE contains one or two intersubunit disulphide linkages is considered.


The active center of a mammalian alpha-amylase. Structure of the complex of a pancreatic alpha-amylase with a carbohydrate inhibitor refined to 2.2-A resolution.
An X-ray structure analysis of a crystal of pig pancreatic alpha-amylase (EC 18.104.22.168) that was soaked with acarbose (a pseudotetrasaccharide alpha-amylase inhibitor) showed electron density corresponding to five fully occupied subsites in the active site. The crystal structure was refined to an R-factor of 15.3%, with a root mean square deviation in bond distances of 0.015 A. The model includes all 496 residues of the enzyme, one calcium ion, one chloride ion, 393 water molecules, and five bound sugar rings. The pseudodisaccharide acarviosine that is the essential structural unit responsible for the activity of all inhibitors of the acarbose type was located at the catalytic center. The carboxylic oxygens of the catalytically competent residues Glu233 and Asp300 form hydrogen bonds with the "glycosidic" NH group of the acarviosine group. The third residue of the catalytic triad Asp197 is located on the opposite side of the inhibitor binding cleft with one of its carbonyl oxygens at a 3.3-A distance from the anomeric carbon C-1 of the inhibitor center. Binding of inhibitor induces structural changes at the active site of the enzyme. A loop region between residues 304 and 309 moves in toward the bound saccharide, the resulting maximal mainchain movement being 5 A for His305. The side chain of residue Asp300 rotates upon inhibitor binding and makes strong van der Waals contacts with the imidazole ring of His299. Four histidine residues (His101, His201, His299, and His305) are found to be hydrogen-bonded with the inhibitor. Many protein-inhibitor hydrogen bond interactions are observed in the complex structure, as is clear hydrophobic stacking of aromatic residues with the inhibitor surface. The chloride activator ion and structural calcium ion are hydrogen-bonded via their ligands and water molecules to the catalytic residues.


Developmentally regulated expression of adrenal 17 alpha-hydroxylase cytochrome P450 in the mouse embryo.
Corticosterone is the major circulating glucocorticoid in adult mice and rats, and this is explained, in part, by the absence of 17 alpha-hydroxylase cytochrome P450 (P450c17) in adrenal glands of these rodents. During embryonic development, however, we discovered transient expression of P450c17 in a subset of adrenocortical cells in the fetal mouse adrenal. This differs from cholesterol side-chain cleavage cytochrome P450 and adrenodoxin, which are expressed continuously in most fetal adrenocortical cells from onset of expression at embryonic days 11-12 (E11-12) until term. Adrenal P450c17 transcripts are detectable in situ at E12.5 and increase in abundance from E12.5 to E14.5. Transcripts are then lost between E16.5 and term (E18.5) and are undetectable in situ in adrenal glands of adult mice. These results are consistent with the presence of pregnenolone 17 alpha-hydroxylase activity in adrenal homogenates of fetal but not adult mice. By using polymerase chain reaction, we determined that murine fetal (E14.5-15.5) adrenal glands contain messenger RNAs (mRNAs) encoding all of the steroid hydroxylases required to produce cortisol and corticosterone but little aldosterone synthase mRNA. Adrenal glands from adult mice contain mRNAs encoding steroid hydroxylases required to produce corticosterone and aldosterone but not cortisol (little P450c17 mRNA). The spatial and temporal expression patterns of P450c17 and aldosterone synthase mRNA, which differ from those of cholesterol side-chain cleavage cytochrome P450 and adrenodoxin, suggest that multiple factors must be required to program cell type- and species-specific expression of these steroid hydroxylases during embryonic development.


Predictors of disability among midlife men and women: differences by severity of impairment.
This study examines the influences on disability among men and women controlling for the degree of functional incapacity imposed by chronic illness or impairment. The associations between demographic, attitude, social role characteristics and disability are separately examined for persons with mild and severe levels of limitation in order to determine whether different factors are associated with disability when impairment is mild as opposed to severe. Separate logit regressions are estimated for four sex/severity groups using data from the 1978 Survey of Disability and Work. It is hypothesized that among persons with severe impairments, only demographic variables will be significantly associated with disability because the severity of limitations would make other factors unimportant, while among persons who are mildly impaired attitudes and social roles will be important influences. Contrary to these expectations, the factors associated with disability are similar among the four sex/severity groups. In all four groups the presence of pain and fatigue strongly influence disability while gender roles are generally not associated with disability. Attitudes regarding the importance of work for self-esteem are also found to be associated with reports of disability. An unexpected outcome of this analysis is an indication of what predicts the absence of disability among severely impaired persons. In particular, persons who report that their sense of self-esteem is tied to work are least likely to report disability. These findings suggest which non-medical factors may contribute to the maintenance of usual work roles and life activities among persons with severe levels of impairment. They also identify variables which may link impairment and disability across all levels of severity.


The impact of social disadvantage in moderate-to-severe chronic kidney disease: an equity-focused systematic review.
It is unclear whether a social gradient in health outcomes exists for people with moderate-to-severe chronic kidney disease (CKD). We critically review the literature for evidence of social gradients in health and investigate the 'suitability' of statistical analyses in the primary studies. In this equity-focused systematic review among adults with moderate-to-severe CKD, factors of disadvantage included gender, race/ethnicity, religion, education, socio-economic status or social capital, occupation and place of residence. Outcomes included access to healthcare, kidney disease progression, cardiovascular events, all-cause mortality and suitability of analyses. Twenty-four studies in the pre-dialysis population and 34 in the dialysis population representing 8.9 million people from 10 countries were included. In methodologically suitable studies among pre-dialysis patients, a significant social gradient was observed in access to healthcare for those with no health insurance and no home ownership. Low income and no home ownership were associated with higher cardiovascular event rates and higher mortality [HR 1.94, 95% confidence interval (CI) 1.27-2.98; HR 1.28, 95% CI 1.04-1.58], respectively. In methodologically suitable studies among dialysis patients, females, ethnic minorities, those with low education, no health insurance, low occupational level or no home ownership were significantly less likely to access cardiovascular healthcare than their more advantaged dialysis counterparts. Low education level and geographic remoteness were associated with higher cardiovascular event rates and higher mortality (HR 1.54, 95% CI 1.01-2.35; HR 1.21, 95% CI 1.08-1.37), respectively. Socially disadvantaged pre-dialysis and dialysis patients experience poorer access to specialist cardiovascular health services, and higher rates of cardiovascular events and mortality than their more advantaged counterparts.


Clinical relevance and IgG subclass determination of non-HLA antibodies identified using endothelial cell precursors isolated from donor blood.
ABO and human leukocyte antigen (HLA) alloantibodies provide major immunologic barriers to successful transplantation; however, there is increasing recognition for the role of anti-endothelial cell antibodies (AECAs) in allograft rejection. We investigated the relationship between AECAs identified using donor-derived endothelial cell precursors (ECPs) and kidney allograft rejection and function. Sixty live donor kidney recipients were tested pretransplant for AECAs and HLA-antibodies using flow cytometric crossmatch tests and solid-phase bead immunoassays. Renal allograft function was assessed by serum creatinine (SCr) values collected at early (mean, 50 days) and late (mean, 815 days) time points posttransplant and by incidence and type of rejection. Immunoglobulin G (IgG) subtype determination of both AECAs and HLA antibodies bound to ECPs was performed using flow cytometry. Fourteen patients (23%) tested positive for donor-reactive IgG AECAs and had statistically higher SCr values and incidences of cellular rejection early posttransplant compared with 46 patients who tested negative (P=0.014 and P<0.05). SCr values were not statistically different late posttransplant. IgG subclass determination showed AECAs to be enriched for IgG2 and IgG4, subclasses that do not activate complement effectively. Detection of donor-reactive immunoglobulin M (IgM) AECAs did not correlate with increased SCr or incidence of rejection. Crossmatch tests performed using donor-derived ECPs allow for the identification of alloantibodies that are associated with cellular rejection and are distinct from alloantibodies detected using lymphocytes.


Human Ecstasy use is associated with increased cortical excitability: an fMRI study.
The serotonergic neurotoxin, 3,4-methylenedioxymethamphetamine (MDMA/Ecstasy), is a highly popular recreational drug. Human recreational MDMA users have neurocognitive and neuropsychiatric impairments, and human neuroimaging data are consistent with animal reports of serotonin neurotoxicity. However, functional neuroimaging studies have not found consistent effects of MDMA on brain neurophysiology in human users. Several lines of evidence suggest that studying MDMA effects in visual system might reveal the general cortical and subcortical neurophysiological consequences of MDMA use. We used 3 T functional magnetic resonance imaging during visual stimulation to compare visual system lateral geniculate nucleus (LGN) and Brodmann Area (BA) 17 and BA 18 activation in 20 long abstinent (479.95±580.65 days) MDMA users and 20 non-MDMA user controls. Lifetime quantity of MDMA use was strongly positively correlated with blood oxygenation level-dependent (BOLD) signal intensity in bilateral LGN (r(s)=0.59; p=0.007), BA 17 (r(s)=0.50; p=0.027), and BA 18 (r(s)=0.48; p=0.031), and with the spatial extent of activation in BA 17 (r(s)=0.059; p=0.007) and BA 18 (r(s)=0.55; p=0.013). There were no between-group differences in brain activation in any region, but the heaviest MDMA users showed a significantly greater spatial extent of activation than controls in BA 17 (p=0.031) and BA 18 (p=0.049). These results suggest that human recreational MDMA use may be associated with a long-lasting increase in cortical excitability, possibly through loss of serotonin input to cortical and subcortical regions. When considered in the context of previous results, cortical hyper-excitability may be a biomarker for MDMA-induced serotonin neurotoxicity.


PCR-based assays for the fish pathogen Aeromonas salmonicida. II. Further evaluation and validation of three PCR primer sets with infected fish.
Two Aeromonas salmonicida-specific polymerase chain reaction (PCR) tests and 1 A. salmonicida subsp. salmonicida-specific PCR test were used to screen salmonid populations that were either overtly or covertly infected with A. salmonicida subsp. salmonicida. It was demonstrated that these PCR assays could be used to replace the biochemical testing currently employed to confirm the identity of A. salmonicida isolates cultured from infected fish. The AP and PAAS PCR assays were also capable of direct detection of A. salmonicida in overtly infected fish, with mucus, gill and kidney samples most likely to yield a positive result. Culture was a more reliable method for the direct detection of A. salmonicida in covertly infected salmonids than was the direct PCR testing of tissue samples, with the AP and PAAS PCRs having a lower detection limit (LDL) of approximately 4 x 10(5) colony-forming units (CFU) g(-1) sample.


Recent extinctions disturb path to equilibrium diversity in Caribbean bats.
Islands are ideal systems to model temporal changes in biodiversity and reveal the influence of humans on natural communities. Although theory predicts biodiversity on islands tends towards an equilibrium value, the recent extinction of large proportions of island biotas complicates testing this model. The well-preserved subfossil record of Caribbean bats-involving multiple insular radiations-provides a rare opportunity to model diversity dynamics in an insular community. Here, we reconstruct the diversity trajectory in noctilionoid bats of the Greater Antilles by applying a dynamic model of colonization, extinction and speciation to phylogenetic and palaeontological data including all known extinct and extant species. We show species richness asymptotes to an equilibrium value, a demonstration of natural equilibrium dynamics across an entire community. However, recent extinctions-many caused by humans-have wiped out nearly a third of island lineages, dragging diversity away from equilibrium. Using a metric to measure island biodiversity loss, we estimate it will take at least eight million years to regain pre-human diversity levels. Our integrative approach reveals how anthropogenic extinctions can drastically alter the natural trajectory of biological communities, resulting in evolutionary disequilibrium.


Adhesion phenotype manifests an altered metabolic profile favoring glycolysis.
To determine whether metabolic markers are differentially expressed in normal and adhesion fibroblasts with and without hypoxia exposure. Prospective experimental study. University research laboratory. Fibroblasts established from normal peritoneum and adhesion tissues from the same patients. In vitro experiments on normal peritoneal and adhesion fibroblasts under normal and hypoxic (2% O2) conditions. Expression of metabolic markers, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), glucose transporter 1 (GLUT1), hypoxia inducible factor (HIF)-1α, hexokinase 2 (HK2), lactose dehydrogenase A (LDHA), and pyruvate dehydrogenase alpha 1 (PDHA1) were measured using real-time reverse transcription polymerase chain reaction; adenosine triphosphate (ATP), HIF-1α, and lactate levels were assessed with ELISAs. Baseline mRNA levels of GAPDH and HIF-1α were increased, while GLUT1 and PDHA1 were decreased in adhesion as compared with in normal peritoneal fibroblasts. There was no change in baseline levels of HK2 or LDHA between the cell lines. Hypoxia increased protein levels of HIF-1α and mRNA levels of GAPDH, GLUT1, and HK2 and decreased levels of PDHA1 in both cell lines. Hypoxia increased LDHA mRNA levels in normal peritoneal fibroblasts. Baseline levels of lactate and ATP were lower in adhesion as compared with in normal peritoneal fibroblasts. In response to hypoxia, there was an increase in lactate in both cell lines and a decrease in ATP in normal fibroblasts. Adhesion fibroblasts manifested an altered metabolic profile, which favors the glycolytic pathway, and is further altered by hypoxia. Targeting these specific metabolic markers during surgery can be an important therapeutic intervention minimizing the development of postoperative adhesions.


Neurogenic mechanisms underlying the rapid onset of sympathetic responses to intermittent hypoxia.
Sleep apnea (SA) leads to metabolic abnormalities and cardiovascular dysfunction. Rodent models of nocturnal intermittent hypoxia (IH) are used to mimic arterial hypoxemias that occur during SA. This mini-review focuses on our work examining central nervous system (CNS) mechanisms whereby nocturnal IH results in increased sympathetic nerve discharge (SND) and hypertension (HTN) that persist throughout the 24-h diurnal period. Within the first 1-2 days of IH, arterial pressure (AP) increases even during non-IH periods of the day. Exposure to IH for 7 days biases nucleus tractus solitarius (NTS) neurons receiving arterial chemoreceptor inputs toward increased discharge, providing a substrate for persistent activation of sympathetic outflow. IH HTN is blunted by manipulations that reduce angiotensin II (ANG II) signaling within the forebrain lamina terminalis suggesting that central ANG II supports persistent IH HTN. Inhibition of the hypothalamic paraventricular nucleus (PVN) reduces ongoing SND and acutely lowers AP in IH-conditioned animals. These findings support a role for the PVN, which integrates information ascending from NTS and descending from the lamina terminalis, in sustaining IH HTN. In summary, our findings indicate that IH rapidly and persistently activates a central circuit that includes the NTS, forebrain lamina terminalis, and the PVN. Our working model holds that NTS neuromodulation increases transmission of arterial chemoreceptor inputs, increasing SND via connections with PVN and rostral ventrolateral medulla. Increased circulating ANG II sensed by the lamina terminalis generates yet another excitatory drive to PVN. Together with adaptations intrinsic to the PVN, these responses to IH support rapid onset neurogenic HTN.


Effect of 1 year daily treatment with 400 microg budesonide (Pulmicort Turbuhaler) in newly diagnosed asthmatics.
The aim of this study was to investigate whether treatment with a low daily dose of 400 microg inhaled budesonide (Pulmicort Turbuhaler) in newly diagnosed asthmatics could influence the course of asthma. Seventy five adult patients, mostly with mild asthma, diagnosed during the previous year and bronchial hyperresponsiveness, participated in a double-blind, randomized, parallel-group multicentre study. They were treated with budesonide 200 microg b.i.d. or placebo, delivered via Turbuhaler for 12 months and followed-up for another 6 months without inhaled steroid treatment. Airway function, symptom scores, reactivity to histamine and inflammatory indices in blood were assessed. The mean increase in morning peak expiratory flow (PEF) was 28 L x min(-1) after budesonide treatment compared with no increase in the placebo group (p=0.011). The provocative dose of histamine causing a 20% fall in forced expiratory volume in one second (PD20) (geometric mean) increased in the budesonide group by approximately two doubling dose steps, but not in the placebo group (p=0.0003). The difference between groups with regard to improvement in asthma symptom scores and inflammatory indices did not reach statistical significance. During the 6 month follow-up, the PEF values of the patients who had previously been treated with budesonide decreased by 18 L x min(-1) while the PD20 decreased by approximately one doubling dose step. In conclusion, early treatment with a low dose of budesonide improves airway function and decreases bronchial responsiveness, but the improvements are short-lasting without continued treatment.


Caregivers of frail elders: updating a national profile.
Family caregivers comprise the backbone of long-term-care provision in the United States, yet little is known about how the composition and experience of family caregiving has changed over time. Data are drawn from the 1989 and 1999 National Long-Term Care Survey and Informal Caregiver Survey to develop nationally representative profiles of disabled older adults and their primary informal caregivers at two points in time. The proportion of chronically disabled community-dwelling older adults who were receiving informal assistance from family or friends declined over the period of interest, whereas the proportion receiving no human help increased. On average, recipients of informal care were older and more disabled in 1999 than in 1989. Primary caregivers were children (41.3%), spouses (38.4%), and other family or friends (20.4%); children were more likely and others less likely to serve as primary caregivers in 1999 relative to 1989. Primary caregivers provided frequent and high levels of help at both points in time. A striking increase was found (from 34.9% to 52.8%) in the proportion of primary caregivers working alone, without secondary caregiver involvement. In the context of projected demographic trends and budgetary constraints to public health insurance programs, these data underscore the importance of identifying viable strategies to monitor and support family caregivers in the coming years.


Hepatitis E: prevention and treatment.
Infection with the hepatitis E virus (HEV) is very common worldwide. The epidemiology, viral genotypes, and transmission routes differ between low-resource countries and economically developed countries. These differences have resulted in the design of diverse prevention and treatment strategies to combat HEV. The population seroprevalence of HEV immunoglobulin G varies between 5 and 50%. However, the diagnosis of acute hepatitis from HEV has not been common in the United States or Western Europe. Chronic progressive HEV infections have been reported among patients who are immunocompromised. Successful treatment of patients with chronic hepatitis from HEV infection with antiviral agents, such as ribavirin or interferon-α, has been reported. Extrahepatic manifestations of HEV infection are common. Large epidemics of hundreds or thousands of cases continue to be reported among populations in Asia and Africa. A subunit peptide HEV vaccine has been found to be highly efficacious in a large clinical trial. However, the vaccine has not been evaluated in populations of pregnant women or other risk groups and is only available in China. Although HEV infections are increasingly recognized as a global public health problem, there are few methods for prevention and treatment that are widely available.


Optimal ambulance location with random delays and travel times.
We describe an ambulance location optimization model that minimizes the number of ambulances needed to provide a specified service level. The model measures service level as the fraction of calls reached within a given time standard and considers response time to be composed of a random delay (prior to travel to the scene) plus a random travel time. In addition to modeling the uncertainty in the delay and in the travel time, we incorporate uncertainty in the ambulance availability in determining the response time. Models that do not account for the uncertainty in all three of these components may overestimate the possible service level for a given number of ambulances and underestimate the number of ambulances needed to provide a specified service level. By explicitly modeling the randomness in the ambulance availability and in the delays and the travel times, we arrive at a more realistic ambulance location model. Our model is tractable enough to be solved with general-purpose optimization solvers for cities with populations around one Million. We illustrate the use of the model using actual data from Edmonton.


Linking single-cell measurements of mass, growth rate, and gene expression.
Mass and growth rate are highly integrative measures of cell physiology not discernable via genomic measurements. Here, we introduce a microfluidic platform enabling direct measurement of single-cell mass and growth rate upstream of highly multiplexed single-cell profiling such as single-cell RNA sequencing. We resolve transcriptional signatures associated with single-cell mass and growth rate in L1210 and FL5.12 cell lines and activated CD8+ T cells. Further, we demonstrate a framework using these linked measurements to characterize biophysical heterogeneity in a patient-derived glioblastoma cell line with and without drug treatment. Our results highlight the value of coupled phenotypic metrics in guiding single-cell genomics.


Cockayne syndrome-derived neurons display reduced synapse density and altered neural network synchrony.
Cockayne syndrome (CS) is a rare genetic disorder in which 80% of cases are caused by mutations in the Excision Repair Cross-Complementation group 6 gene (ERCC6). The encoded ERCC6 protein is more commonly referred to as Cockayne Syndrome B protein (CSB). Classical symptoms of CS patients include failure to thrive and a severe neuropathology characterized by microcephaly, hypomyelination, calcification and neuronal loss. Modeling the neurological aspect of this disease has proven difficult since murine models fail to mirror classical neurological symptoms. Therefore, a robust human in vitro cellular model would advance our fundamental understanding of the disease and reveal potential therapeutic targets. Herein, we successfully derived functional CS neural networks from human CS induced pluripotent stem cells (iPSCs) providing a new tool to facilitate studying this devastating disease. We identified dysregulation of the Growth Hormone/Insulin-like Growth Factor-1 (GH/IGF-1) pathway as well as pathways related to synapse formation, maintenance and neuronal differentiation in CSB neurons using unbiased RNA-seq gene expression analyses. Moreover, when compared to unaffected controls, CSB-deficient neural networks displayed altered electrophysiological activity, including decreased synchrony, and reduced synapse density. Collectively, our work reveals that CSB is required for normal neuronal function and we have established an alternative to previously available models to further study neural-specific aspects of CS.


An evolutionary model-based algorithm for accurate phylogenetic breakpoint mapping and subtype prediction in HIV-1.
Genetically diverse pathogens (such as Human Immunodeficiency virus type 1, HIV-1) are frequently stratified into phylogenetically or immunologically defined subtypes for classification purposes. Computational identification of such subtypes is helpful in surveillance, epidemiological analysis and detection of novel variants, e.g., circulating recombinant forms in HIV-1. A number of conceptually and technically different techniques have been proposed for determining the subtype of a query sequence, but there is not a universally optimal approach. We present a model-based phylogenetic method for automatically subtyping an HIV-1 (or other viral or bacterial) sequence, mapping the location of breakpoints and assigning parental sequences in recombinant strains as well as computing confidence levels for the inferred quantities. Our Subtype Classification Using Evolutionary ALgorithms (SCUEAL) procedure is shown to perform very well in a variety of simulation scenarios, runs in parallel when multiple sequences are being screened, and matches or exceeds the performance of existing approaches on typical empirical cases. We applied SCUEAL to all available polymerase (pol) sequences from two large databases, the Stanford Drug Resistance database and the UK HIV Drug Resistance Database. Comparing with subtypes which had previously been assigned revealed that a minor but substantial (approximately 5%) fraction of pure subtype sequences may in fact be within- or inter-subtype recombinants. A free implementation of SCUEAL is provided as a module for the HyPhy package and the Datamonkey web server. Our method is especially useful when an accurate automatic classification of an unknown strain is desired, and is positioned to complement and extend faster but less accurate methods. Given the increasingly frequent use of HIV subtype information in studies focusing on the effect of subtype on treatment, clinical outcome, pathogenicity and vaccine design, the importance of accurate, robust and extensible subtyping procedures is clear.


Different APC genotypes in proximal and distal sporadic colorectal cancers suggest distinct WNT/β-catenin signalling thresholds for tumourigenesis.
Biallelic protein-truncating mutations in the adenomatous polyposis coli (APC) gene are prevalent in sporadic colorectal cancer (CRC). Mutations may not be fully inactivating, instead producing WNT/β-catenin signalling levels 'just-right' for tumourigenesis. However, the spectrum of optimal APC genotypes accounting for both hits, and the influence of clinicopathological features on genotype selection remain undefined. We analysed 630 sporadic CRCs for APC mutations and loss of heterozygosity (LOH) using sequencing and single-nucleotide polymorphism microarrays, respectively. Truncating APC mutations and/or LOH were detected in 75% of CRCs. Most truncating mutations occurred within a mutation cluster region (MCR; codons 1282-1581) leaving 1-3 intact 20 amino-acid repeats (20AARs) and abolishing all Ser-Ala-Met-Pro (SAMP) repeats. Cancers commonly had one MCR mutation plus either LOH or another mutation 5' to the MCR. LOH was associated with mutations leaving 1 intact 20AAR. MCR mutations leaving 1 vs 2-3 intact 20AARs were associated with 5' mutations disrupting or leaving intact the armadillo-repeat domain, respectively. Cancers with three hits had an over-representation of mutations upstream of codon 184, in the alternatively spliced region of exon 9, and 3' to the MCR. Microsatellite unstable cancers showed hyper-mutation at MCR mono- and di-nucleotide repeats, leaving 2-3 intact 20AARs. Proximal and distal cancers exhibited different preferred APC genotypes, leaving a total of 2 or 3 and 0 to 2 intact 20AARs, respectively. In conclusion, APC genotypes in sporadic CRCs demonstrate 'fine-tuned' interdependence of hits by type and location, consistent with selection for particular residual levels of WNT/β-catenin signalling, with different 'optimal' thresholds for proximal and distal cancers.


Preparing bulk ultrafine-microstructure high-entropy alloys via direct solidification.
In the past three decades, nanostructured (NS) and ultrafine-microstructure (UFM) materials have received extensive attention due to their excellent mechanical properties such as high strength. However, preparing low-cost and bulk NS and UFM materials remains to be a challenge, which limits their industrial applications. Here, we report a new strategy to prepare bulk UFM alloys via the direct solidification of high-entropy alloys (HEAs). As a proof of concept, we designed AlCoCrxFeNi (1.8 ≤ x ≤ 2.0) HEAs and achieved a complete UFM in bulk materials. The compositional requirements for obtaining the formation of the UFM are highly demanding, necessitating the coupling of near eutectic alloy composition and the high temperature decomposition of supersaturated primary and secondary phases. Our strategy provides a low-cost and highly efficient method to prepare bulk UFM alloys, with great potential to accelerate the engineering application of these materials.


Posterior crossbite--treatment and stability.
Posterior crossbite is defined as an inadequate transversal relationship of maxillary and mandibular teeth. Even when eliminating the etiologic factors, this malocclusion does not have a spontaneous correction, and should be treated with maxillary expansion as early as possible. This treatment aims at providing a better tooth/skeletal relationship, thereby improving masticatory function, and establishing a symmetrical condyle/fossa relationship. Should posterior crossbite not be treated early, it may result in skeletal changes, demanding a more complex approach. Additionally, an overcorrection expansion protocol should be applied in order to improve the treatment stability. Although the literature has reported a high rate of relapse after maxillary expansion, the goal of this study was to demonstrate excellent stability of the posterior crossbite correction 21 years post treatment.


Plasmodium hemozoin formation mediated by histidine-rich proteins.
The digestive vacuole of Plasmodium falciparum is the site of hemoglobin degradation, heme polymerization into crystalline hemozoin, and antimalarial drug accumulation. Antibodies identified histidine-rich protein II (HRP II) in purified digestive vacuoles. Recombinant or native HRP II promoted the formation of hemozoin, and chloroquine inhibited the reaction. The related HRP III also polymerized heme, and an additional HRP was identified in vacuoles. It is proposed that after secretion by the parasite into the host erythrocyte cytosol, HRPs are brought into the acidic digestive vacuole along with hemoglobin. After hemoglobin proteolysis, HRPs bind the liberated heme and mediate hemozoin formation.


The Risk of Gout Among Patients With Sleep Apnea: A Matched Cohort Study.
Obstructive sleep apnea (OSA) is associated with a range of serious comorbidities. This study was undertaken to investigate whether people with OSA are more likely to develop gout, in the short and long term, compared to those without OSA. A matched retrospective cohort study was undertaken using the UK Clinical Practice Research Datalink. Individuals age ≥18 years who received a diagnosis of OSA between 1990 and 2010 were identified and matched on age, sex, and practice with up to 4 individuals without OSA; follow-up was until the end of 2015. Hazard ratios (HRs) were estimated using Cox regression adjusted for general health, lifestyle, and comorbidity characteristics. The risk of developing gout was assessed at different time points, and the body mass index (BMI) category-specific results were presented. The study sample included 15,879 patients with OSA and 63,296 without. The median follow-up was 5.8 years. We found that 4.9% of patients with OSA and 2.6% of patients without the disorder developed gout. The incidence rate per 1,000 person-years was 7.83 (95% confidence interval [95% CI] 7.29-8.40) and 4.03 (95% CI 3.84-4.23) among those with and without OSA, respectively. The adjusted HR was 1.42 (95% CI 1.29-1.56). The risk of developing gout among OSA patients compared to those without was highest 1-2 years after the index date (HR 1.64 [95% CI 1.30-2.06]). This finding persisted among those who were overweight and obese. For those with normal BMI, the highest significant HR (2.02 [95% CI 1.13-3.62]) was observed at 2-5 years after the index date. In this study, patients with OSA continued to be at higher risk of developing gout beyond the first year following the diagnosis. Our results further indicate that peak incidences of gout vary according to BMI.


Mesenchymal regulation of epithelial gene expression in developing avian stomach: 5'-flanking region of pepsinogen gene can mediate mesenchymal influence on its expression.
The expression of a gene encoding an embryonic chick pepsinogen was investigated in developing avian gut. Expression is restricted to the epithelial layer of the embryonic proventriculus (glandular stomach). We can therefore regard this gene as a marker gene for proventricular epithelial differentiation. There is some considerable evidence in favour of epithelial-mesenchymal interactions being important during the development of the gastrointestinal system; for example, pepsinogen expression is induced in proventricular and gizzard (muscular stomach) epithelial by the proventricular mesenchyme but is suppressed by the gizzard mesenchyme. In the present paper, we studied how the mesenchymes influence this gene expression pattern. For this we produced constructs containing various portions of the 5'-flanking region of the embryonic chick pepsinogen gene, driving reporter sequences (beta-galactocidase or luciferase), and these constructs were transfected into dissociated epithelial cells either from the proventriculus or gizzard. We then recombined these cells with mesenchymal cells and cultured them as cell aggregates. In this way, we were able to dissect the timing and other requirements of the epithelial-mesenchymal interactions for expression of embryonic chick pepsinogen gene. We also report that 1.1 kb of 5'-flanking sequence is sufficient to drive correct expression of embryonic chick pepsinogen gene, although further enhancement was seen if the constructs contained 3.2 kb of upstream sequence.


Muscle Activation Patterns Are More Constrained and Regular in Treadmill Than in Overground Human Locomotion.
The use of motorized treadmills as convenient tools for the study of locomotion has been in vogue for many decades. However, despite the widespread presence of these devices in many scientific and clinical environments, a full consensus on their validity to faithfully substitute free overground locomotion is still missing. Specifically, little information is available on whether and how the neural control of movement is affected when humans walk and run on a treadmill as compared to overground. Here, we made use of linear and non-linear analysis tools to extract information from electromyographic recordings during walking and running overground, and on an instrumented treadmill. We extracted synergistic activation patterns from the muscles of the lower limb via non-negative matrix factorization. We then investigated how the motor modules (or time-invariant muscle weightings) were used in the two locomotion environments. Subsequently, we examined the timing of motor primitives (or time-dependent coefficients of muscle synergies) by calculating their duration, the time of main activation, and their Hurst exponent, a non-linear metric derived from fractal analysis. We found that motor modules were not influenced by the locomotion environment, while motor primitives were overall more regular in treadmill than in overground locomotion, with the main activity of the primitive for propulsion shifted earlier in time. Our results suggest that the spatial and sensory constraints imposed by the treadmill environment might have forced the central nervous system to adopt a different neural control strategy than that used for free overground locomotion, a data-driven indication that treadmills could induce perturbations to the neural control of locomotion.


White matter hyperintensities and imaging patterns of brain ageing in the general population.
White matter hyperintensities are associated with increased risk of dementia and cognitive decline. The current study investigates the relationship between white matter hyperintensities burden and patterns of brain atrophy associated with brain ageing and Alzheimer's disease in a large populatison-based sample (n = 2367) encompassing a wide age range (20-90 years), from the Study of Health in Pomerania. We quantified white matter hyperintensities using automated segmentation and summarized atrophy patterns using machine learning methods resulting in two indices: the SPARE-BA index (capturing age-related brain atrophy), and the SPARE-AD index (previously developed to capture patterns of atrophy found in patients with Alzheimer's disease). A characteristic pattern of age-related accumulation of white matter hyperintensities in both periventricular and deep white matter areas was found. Individuals with high white matter hyperintensities burden showed significantly (P < 0.0001) lower SPARE-BA and higher SPARE-AD values compared to those with low white matter hyperintensities burden, indicating that the former had more patterns of atrophy in brain regions typically affected by ageing and Alzheimer's disease dementia. To investigate a possibly causal role of white matter hyperintensities, structural equation modelling was used to quantify the effect of Framingham cardiovascular disease risk score and white matter hyperintensities burden on SPARE-BA, revealing a statistically significant (P < 0.0001) causal relationship between them. Structural equation modelling showed that the age effect on SPARE-BA was mediated by white matter hyperintensities and cardiovascular risk score each explaining 10.4% and 21.6% of the variance, respectively. The direct age effect explained 70.2% of the SPARE-BA variance. Only white matter hyperintensities significantly mediated the age effect on SPARE-AD explaining 32.8% of the variance. The direct age effect explained 66.0% of the SPARE-AD variance. Multivariable regression showed significant relationship between white matter hyperintensities volume and hypertension (P = 0.001), diabetes mellitus (P = 0.023), smoking (P = 0.002) and education level (P = 0.003). The only significant association with cognitive tests was with the immediate recall of the California verbal and learning memory test. No significant association was present with the APOE genotype. These results support the hypothesis that white matter hyperintensities contribute to patterns of brain atrophy found in beyond-normal brain ageing in the general population. White matter hyperintensities also contribute to brain atrophy patterns in regions related to Alzheimer's disease dementia, in agreement with their known additive role to the likelihood of dementia. Preventive strategies reducing the odds to develop cardiovascular disease and white matter hyperintensities could decrease the incidence or delay the onset of dementia.


Adherence of patients to long-term medication: a cross-sectional study of antihypertensive regimens in Austria.
The objective of this study was to evaluate adherence and causes for non-adherence to antihypertensive therapy in Austrian patients. A special focus was placed on social parameters and behavioural theories. Patients were invited via advertisements in community pharmacies in Austria to complete an online survey. Inclusion criteria were an age of 18 years or older, a diagnosis of arterial hypertension and a current prescription of antihypertensive medication. Adherence was measured by the four-item Morisky scale. Non-adherence was defined by at least one point in the Morisky scale. Several demographic, social and behavioural parameters were analysed as potential co-variables associated with adherence. A total of 323 patients completed the online survey, of which 109 (33.7%) met the criteria for non-adherence. In a multivariable model, self-efficacy and age were associated with adherence, whereas intention and barriers were linked to non-adherence; 56 patients (17.3%) were classified as intentionally non-adherent. This study demonstrates that non-adherence affects an important proportion of patients in the treatment of arterial hypertension. Young age was a particularly important risk factor for non-adherence, and this patient population is, therefore, in need of special attention. Modifiable risk factors were identified that could help improving the treatment of arterial hypertension and potentially other chronic conditions.


Bayesian modeling of embryonic growth using latent variables.
In a growth model, individuals move progressively through a series of states in which each state is indicative of developmental status. Interest lies in estimating the rate of progression through each state while incorporating covariates that might affect the transition rates. We develop a Bayesian discrete-time multistate growth model for inference from cross-sectional data with unknown initiation times. For each subject, data are collected at only one time point at which we observe the state as well as covariates that measure developmental progress. We link the developmental progress variables to an underlying latent growth variable that can also affect the state transition rates. A subject with slow latent growth will then have relatively small developmental progress covariates and move through state transitions slowly. We then examine the association between latent growth and the probability of future events in a novel study of embryonic development and pregnancy loss. Using a Markov chain Monte Carlo (MCMC) algorithm for posterior computation, we found evidence in favor of a previously hypothesized but unproven association between slow growth early in pregnancy and increased risk of future spontaneous abortion.


Is the Relationship Between Pornography Consumption Frequency and Lower Sexual Satisfaction Curvilinear? Results From England and Germany.
Several studies using different methods have found that pornography consumption is associated with lower sexual satisfaction. The language used by media-effects scholars in discussions of this association implies an expectation that lowered satisfaction is primarily due to frequent-but not infrequent-consumption. Actual analyses, however, have assumed linearity. Linear analyses presuppose that for each increase in the frequency of pornography consumption there is a correspondingly equivalent decrease in sexual satisfaction. The present brief report explored the possibility that the association is curvilinear. Survey data from two studies of heterosexual adults, one conducted in England and the other in Germany, were employed. Results were parallel in each country and were not moderated by gender. Quadratic analysis indicated a curvilinear relationship, in the form of a predominantly negative, concave downward curve. Simple slope analyses suggested that when the frequency of consumption reaches once a month, sexual satisfaction begins to decrease, and that the magnitude of the decrease becomes larger with each increase in the frequency of consumption. The observational nature of the data employed precludes any causal inferences. However, if an effects perspective was adopted, these results would suggest that low rates of pornography consumption have no impact on sexual satisfaction and that adverse effects initiate only after consumption reaches a certain frequency.


Avoidance of posterior tectal membranes by temporal retinal axons.
Membrane carpets consisting of alternating membrane stripes were prepared from plasma membranes of anterior and posterior chick optic tectum. Axons from retinal explants extend neurites on these carpets. Axons of the nasal retina do not distinguish between the stripes. Axons of the temporal retina prefer to extend neurites on anterior tectal membranes. Treatment of the membrane fragments with high temperature interferes with the pattern of neurite outgrowth from temporal axons. When growing on carpets consisting of treated anterior and posterior tectal membranes, temporal retinal axons no longer distinguish between the stripes. Treatment of posterior membranes alone is sufficient to abolish the preference of temporal axons to extend neurites on anterior tectal membranes. Treatment of the anterior membranes alone has no effect. This result is best explained by a repulsive component in the posterior tectal membranes. Temporal, but not nasal, axons specifically recognize and avoid that component, with the result that they do not extend neurites on posterior tectal membrane stripes. Once the repulsive component is destroyed, temporal axons are able to extend neurites on posterior tectal membranes.


Natural resistance to Clover yellow vein virus in beans controlled by a single recessive locus.
We characterized the resistance of the common bean cv. Jolanda to Clover yellow vein virus no. 30 (ClYVV). After inoculation, the virus was detected in neither inoculated nor upper leaves, suggesting that the resistance operates at either the viral replication or cell-to-cell movement level. To analyze the mechanism of resistance, we developed a green fluorescent protein (GFP)-tagged ClYVV, and monitored GFP fluorescence at sites of infection on ClYVV-inoculated leaves. No GFP fluorescence was detected in Jolanda, whereas its expression in single cells and spread on inoculated leaves were observed clearly in susceptible cultivars. ClYVV-introduced Jolanda cells were found to be still viable; therefore, it is unlikely that the restriction of multiplication was due to rapid cell death. Genetic analysis indicated that a single recessive locus controlled the resistant phenotype of Jolanda. We designated this locus desc (determinant of susceptibility to ClYVV). Meanwhile, a spontaneous mutant virus that overcomes the resistance (ClYVV-Br) was isolated. Inoculation assays using chimeric viruses suggested that a viral genome-linked protein (VPg) might be the avirulence determinant. The resistance mechanism may be associated with the role of VPg in the viral infection cycle.


Free-flow zone electrophoresis and isoelectric focusing using a microfabricated glass device with ion permeable membranes.
This paper describes a microfabricated free-flow electrophoresis device with integrated ion permeable membranes. In order to obtain continuous lanes of separated components an electrical field is applied perpendicular to the sample flow direction. This sample stream is sandwiched between two sheath flow streams, by hydrodynamic focusing. The separation chamber has two open side beds with inserted electrodes to allow ventilation of gas generated during electrolysis. To hydrodynamically isolate the separation compartment from the side electrodes, a photo-polymerizable monomer solution is exposed to UV light through a slit mask for in situ membrane formation. These so-called salt-bridges resist the pressure driven fluid, but allow ion transport to enable electrical connection. In earlier devices the same was achieved by using open side channel arrays. However, only a small fraction of the applied voltage was effectively utilized across the separation chamber during free-flow electrophoresis and free-flow isoelectric focusing. Furthermore, the spreading of the carrier ampholytes into the side channels resulted in a very restricted pH gradient inside the separation chamber. The chip presented here allows at least 10 times more efficient use of the applied potential and a nearly linear pH gradient from pH 3 to 10 during free-flow isoelectric focusing could be established. Furthermore, the application of hydrodynamic focusing in combination with free-flow electrophoresis can be used for guiding the separated components to specific chip outlets. As a demonstration, several standard fluorescent markers were separated and focused by free-flow zone electrophoresis and by free-flow isoelectric focusing employing a transversal voltage of up to 150 V across the separation chamber.


Does influenza A virus infection affect movement behaviour during stopover in its wild reservoir host?
The last decade has seen a surge in research on avian influenza A viruses (IAVs), in part fuelled by the emergence, spread and potential zoonotic importance of highly pathogenic virus subtypes. The mallard (Anas platyrhynchos) is the most numerous and widespread dabbling duck in the world, and one of the most important natural hosts for studying IAV transmission dynamics. In order to predict the likelihood of IAV transmission between individual ducks and to other hosts, as well as between geographical regions, it is important to understand how IAV infection affects the host. In this study, we analysed the movements of 40 mallards equipped with GPS transmitters and three-dimensional accelerometers, of which 20 were naturally infected with low pathogenic avian influenza virus (LPAIV), at a major stopover site in the Northwest European flyway. Movements differed substantially between day and night, as well as between mallards returning to the capture site and those feeding in natural habitats. However, movement patterns did not differ between LPAIV infected and uninfected birds. Hence, LPAIV infection probably does not affect mallard movements during stopover, with high possibility of virus spread along the migration route as a consequence.


Motivating medical students by utilizing dermatology-oriented online quizzes.
Online quiz competitions can facilitate extra-classroom interactions between faculty and medical students. Owing to decreased class attendance nationwide, teaching faculty might revamp their approach to medical education by providing online resources and methods for communication. To explore if the use of online quizzes and social media can result in improved interactions between faculty and students. A pilot study conducted from April 7th, 2015 to June 11th, 2015 at Northeast Ohio Medical University (NEOMED) among participants from the second year medical school class. Ten one-question quizzes created using Google Forms were announced over Twitter and email at the rate of one quiz per week. The first correct responder of each quiz chose a five-dollar coffee shop gift card, movie ticket, or a meeting with a NEOMED faculty person as their prize. An average of 23.8% of the second year medical student class at NEOMED participated per quiz. A total of 80 individuals (55.9%) submitted 340 responses during the competition. This is a single-center study with a limited sample size. This study presents a process evaluation for the use of online quiz competitions amongst medical students. Optional online quizzes with small incentives may foster motivational competition among medical students, increase online interactions with faculty, and serve as study material for exams.