[Health technology assessment report: Computer-assisted Pap test for cervical cancer screening].
HEALTH PROBLEM: Cervical cancer is a disease which is highly preventable by means of Pap test screening for the precancerous lesions, which can be easily treated. Furthermore, in the near future, control of the disease will be enhanced by the vaccination which prevents the infection of those human papillomavirus types that cause the vast majority of cervical cancers. The effectiveness of screening in drastically reducing cervical cancer incidence has been clearly demonstrated. The epidemiology of cervical cancer in industrialised countries is now determined mostly by the Pap test coverage of the female population and by the ability of health systems to assure appropriate follow up after an abnormal Pap test. Today there are two fully automated systems for computer-assisted Pap test: the BD FocalPoint and the Hologic Imager. Recently, the Hologic Integrated Imager, a semi-automated system, was launched. The two fully automated systems are composed of a central scanner, where the machine examines the cytologic slide, and of one or more review stations, where the cytologists analyze the slides previously centrally scanned. The softwares used by the two systems identify the fields of interest so that the cytologists can look only at those points, automatically pointed out by the review station. Furthermore, the FocalPoint system classifies the slides according to their level of risk of containing signs of relevant lesions. Those in the upper classes--about one fifth of the slides--are labelled as « further review », while those in the lower level of risk, i.e. slides that have such a low level of risk that they can be considered as negative with no human review, are labelled as « no further review ». The aim of computer-assisted Pap test is to reduce the time of slide examination and to increase productivity. Furthermore, the number of errors due to lack of attention may decrease. Both the systems can be applied to liquidbased cytology, while only the BD Focal Point can be used on conventional smears. Cytology screening has some critical points: there is a shortage of cytologists/cytotechnicians; the quality strongly depends on the experience and ability of the cytologist; there is a subjective component in the cytological diagnosis; in highly screened populations, the prevalence of lesions is very low and the activity of cytologists is very monotonous. On the other hand, a progressive shift to molecular screening using HPV-DNA test as primary screening test is very likely in the near future; cytology will be used as triage test, dramatically reducing the number of slides to process and increasing the prevalence of lesions in those Pap tests. In this Report we assume that the diagnostic accuracy of computer-assisted Pap test is equal to the accuracy of manual Pap test and, consequently, that screening using computer-assisted Pap test has the same efficacy in reducing cervical cancer incidence and mortality. Under this assumption, the effectiveness/ benefit/utility is the same for the two screening modes, i.e. the economic analysis will be a cost minimization study. Furthermore, the screening process is identical for the two modalities in all the phases except for slide interpretation. The cost minimization analysis will be limited to the only phase differing between the two modes, i.e. the study will be a differential cost analysis between a labour-intensive strategy (traditional Pap test) and a technology-intensive strategy (the computer-assisted Pap test). Briefly, the objectives of this HTA Report are: to determine the break even point of computer-assisted Pap test systems, i.e. the volume of slides processed per year at which putting in place a computer-assisted Pap test system becomes economically convenient; to quantify the cost per Pap test in different scenarios according to screening centre activity volume, productivity of cytologist, type of cytology (conventional smear or liquid-based, fully automated or semi-automated computer-assisted); to analyse the computer-assisted Pap test in the Italian context, through a survey of the centres using the technology, collecting data useful for the sensitivity analysis of the economic evaluation; to evaluate the acceptability of the technology in the screening services; to evaluate the organizational and financial impact of the computer-assisted Pap test in different scenarios; to illustrate the ideal organization to implement computer-assisted Pap test in terms of volume of activity, productivity, and human and technological resources. to produce this Report, the following process was adopted: application to the Ministry of health for a grant « Analysis of the impact of professional involvement in evidence generation for the HTA process »; within this project, the sub-project « Cost effectiveness evaluation of the computer-assisted Pap test in the Italian screening programmes » was financed; constitution of the Working Group, which included the project coordinator, the principal investigator, and the health economist; identification of the centres using the computer-assisted Pap test and which had published scientific reports on the subject; identification of the Consulting Committee (stakeholder), which included screening programmes managers, pathologists, economists, health policy-makers, citizen organizations, and manufacturers. Once the evaluation was concluded, a plenary meeting with Working Group and Consulting Committee was held. The working group drafted the final version of this Report, which took into account the comments received. the fully automated computer-assisted Pap test has an important financial and organizational impact on screening programmes. The assessment of this health technology reached the following conclusions: according to the survey results, after some distrust, cytologists accepted the use of the machine and appreciated the reduction in interpretation time and the reliability in identifying the fields of interest; from an economic point of view, the automated computer-assisted Pap test can be convenient only with conventional smears if the screening centre has a volume of more than 49,000 slides/year and the cytologist productivity increases about threefold. It must be highlighted that it is not sufficient to adopt the automated Pap test to reach such an increase in productivity; the laboratory must be organised or re-organised to optimise the use of the review stations and the person time. In the case of liquid-based cytology, the adoption of automated computer- assisted Pap test can only increase the costs. In fact, liquid-based cytology increases the cost of consumable materials but reduces the interpretation time, even in manual screening. Consequently, the reduction of human costs is smaller in the case of computer-assisted screening. Liquid-based cytology has other implications and advantages not linked to the use of computer-assisted Pap test that should be taken into account and are beyond the scope of this Report; given that the computer-assisted Pap test reduces human costs, it may be more advantageous where the cost of cytologists is higher; given the relatively small volume of activity of screening centres in Italy, computer-assisted Pap test may be reasonable for a network using only one central scanner and several remote review stations; the use of automated computer-assisted Pap test only for quality control in a single centre is not economically sustainable. In this case as well, several centres, for example at the regional level, may form a consortium to reach a reasonable number of slides to achieve the break even point. Regarding the use of a machine rather than human intelligence to interpret the slides, some ethical issues were initially raised, but both the scientific community and healthcare professionals have accepted this technology. The identification of fields of interest by the machine is highly reproducible, reducing subjectivity in the diagnostic process. The Hologic system always includes a check by the human eye, while the FocalPoint system identifies about one fifth of the slides as No Further Review. Several studies, some of which conducted in Italy, confirmed the reliability of this classification. There is still some resistance to accept the practice of No Further Review. A check of previous slides and clinical data can be useful to make the cytologist and the clinician more confident. Computer-assisted automated Pap test may be introduced only if there is a need to increase the volume of slides screened to cover the screening target population and sufficient human resources are not available. Switching a programme using conventional slides to automatic scanning can only lead to a reduction in costs if the volume of slides per year exceeds 49,000 slides/annum and cytologist productivity is optimised to more than 20,000 slides per year. At a productivity of 15,000 or fewer, the automated computer-assisted Pap test cannot be convenient. Switching from manual screening with conventional slides to automatic scanning with liquid-based cytology cannot generate any economic saving, but the system could increase output with a given number of staff. The transition from manual to computer assisted automated screening of liquid based cytology will not generate savings and the increase in productivity will be lower than that of the switch from manual/conventional to automated/conventional. The use of biologists or pathologists as cytologists is more costly than the use of cytoscreeners. Given that the automated computer-assisted Pap test reduces human resource costs, its adoption in a model using only biologists and pathologists for screening is more economically advantageous. (ABSTRACT TRUNCATED)


Image analysis of peripheral compression artefacts of ThinPrep(®) liquid-based cytology preparations.
ThinPrep (TP), one of the Food and Drug Administration-approved liquid-based cytology (LBC) preparations, is widely used for gynaecological and non-gynaecological cytology samples. A unique physical artefact caused by the compression at the periphery in TP slides has not been adequately evaluated to date. We processed four established tumour cell lines (MKN28, MKN45, KG-1 and NB4) and mononuclear cells isolated from whole blood over Ficoll-Plaque for TP preparations. For this part of the study, we included five normal cervical LBC preparations. We then auto-counted and auto-measured the area, mean grey value and Feret's diameter in both the inner disc and peripheral rim of the preparations by image morphometry. In addition, we compared the distribution of atypical cell groups in the peripheral rim and inner disc of 132 lung aspirates, 80 thyroid aspirates, 212 cerebrospinal fluids (CSFs) and 50 gynaecological samples. The areas and Feret's diameters of the cytoplasm in the peripheral compressed rim area were statistically larger than those of cells in the inner disc. The mean grey values of cells (cytoplasm and nucleus) in the peripheral compression rim were also smaller than those in the inner disc cells, leading to decreases in nuclear and cytoplasmic chromatism. Except for the mean grey values, the differences were not significant in the cervical samples. Cellular morphology may be markedly distorted in the peripheral rim, regardless of cell malignancy, which may lead to the misinterpretation of cells during the screening. Accordingly, cytological diagnosis based on the findings within the peripheral rim should take this phenomenon into account. Compressed cells found in the peripheral rim should be interpreted with caution when TP slides are used for cytopathological diagnosis.


Relationships between nutrition-related knowledge, self-efficacy, and behavior for fifth grade students attending Title I and non-Title I schools.
The Social Cognitive Theory (SCT) is a widely used theory for nutrition education programming. Better understanding the relationships between knowledge, self-efficacy, and behavior among children of various income levels can help to form and improve nutrition programs, particularly for socioeconomically disadvantaged youth. The purpose of this study was to determine the relationships between knowledge, self-efficacy, and behavior among fifth grade students attending Title I (≥40% of students receiving free or reduced school meals) and non-Title I schools (<40% of students receiving free or reduced school meals). A validated survey was completed by 55 fifth grade students from Title I and 122 from non-Title I schools. Differences in knowledge, self-efficacy, and behavior scores between groups were assessed using t test and adjusted for variations between participating schools. Regression analysis was used to determine the relationships between knowledge, self-efficacy, and behavior. In adjusted models, the Title I group had significantly lower scores on several knowledge items and summary knowledge (P = 0.04). The Title I group had significantly lower scores on several behavior variables including intakes of fruits (P = 0.02), vegetables (P = 0.0005), whole grains (P = 0.0003), and lean protein (P = 0.047), physical activity (P = 0.002) and summary behavior (P = 0.001). However the Title I group scored higher on self-efficacy for meal planning (P = 0.04) and choosing healthy snacks (P = 0.036). Both self-efficacy (β = 0.70, P < 0.0001) and knowledge (β = 0.35, P = 0.002) strongly predicted behavior; however, only self-efficacy remained significant in the Title I group (self-efficacy, β = 0.82, P = 0.0003; knowledge, β = 0.11, P = 0.59). Results demonstrate disparities in nutrition knowledge and behavior outcomes between students surveyed from Title I and non-Title I schools, suggesting more resources may be necessary for lower income populations. Findings suggest that future nutrition interventions should focus on facilitating the improvement of children's self-efficacy.


Phenotype MicroArrays as a complementary tool to next generation sequencing for characterization of tree endophytes.
There is an increasing need to calibrate microbial community profiles obtained through next generation sequencing (NGS) with relevant taxonomic identities of the microbes, and to further associate these identities with phenotypic attributes. Phenotype MicroArray (PM) techniques provide a semi-high throughput assay for characterization and monitoring the microbial cellular phenotypes. Here, we present detailed descriptions of two different PM protocols used in our recent studies on fungal endophytes of forest trees, and highlight the benefits and limitations of this technique. We found that the PM approach enables effective screening of substrate utilization by endophytes. However, the technical limitations are multifaceted and the interpretation of the PM data challenging. For the best result, we recommend that the growth conditions for the fungi are carefully standardized. In addition, rigorous replication and control strategies should be employed whether using pre-configured, commercial microwell-plates or in-house designed PM plates for targeted substrate analyses. With these precautions, the PM technique is a valuable tool to characterize the metabolic capabilities of individual endophyte isolates, or successional endophyte communities identified by NGS, allowing a functional interpretation of the taxonomic data. Thus, PM approaches can provide valuable complementary information for NGS studies of fungal endophytes in forest trees.


Magnetic resonance imaging of the time course of hyperpolarized 129Xe gas exchange in the human lungs and heart.
To perform magnetic resonance imaging (MRI), human lung imaging, and quantification of the gas-transfer dynamics of hyperpolarized xenon-129 (HPX) from the alveoli into the blood plasma. HPX MRI with iterative decomposition of water and fat with echo asymmetry and least-square estimation (IDEAL) approach were used with multi-interleaved spiral k-space sampling to obtain HPX gas and dissolved phase images. IDEAL time-series images were then obtained from ten subjects including six normal subjects and four patients with pulmonary emphysema to test the feasibility of the proposed technique for capturing xenon-129 gas-transfer dynamics (XGTD). The dynamics of xenon gas diffusion over the entire lung was also investigated by measuring the signal intensity variations between three regions of interest, including the left and right lungs and the heart using Welch's t test. The technique enabled the acquisition of HPX gas and dissolved phase compartment images in a single breath-hold interval of 8 s. The y-intersect of the XGTD curves were also found to be statistically lower in the patients with lung emphysema than in the healthy group (p < 0.05). This time-series IDEAL technique enables the visualization and quantification of inhaled xenon from the alveoli to the left ventricle with a clinical gradient strength magnet during a single breath-hold, in healthy and diseased lungs. • The proposed hyperpolarized xenon-129 gas and dissolved magnetic resonance imaging technique can provide regional and temporal measurements of xenon-129 gas-transfer dynamics. • Quantitative measurement of xenon-129 gas-transfer dynamics from the alveolar to the heart was demonstrated in normal subjects and pulmonary emphysema. • Comparison of gas-transfer dynamics in normal subjects and pulmonary emphysema showed that the proposed technique appears sensitive to changes affecting the alveoli, pulmonary interstitium, and capillaries.


The functional interplay between protein kinase CK2 and CCA1 transcriptional activity is essential for clock temperature compensation in Arabidopsis.
Circadian rhythms are daily biological oscillations driven by an endogenous mechanism known as circadian clock. The protein kinase CK2 is one of the few clock components that is evolutionary conserved among different taxonomic groups. CK2 regulates the stability and nuclear localization of essential clock proteins in mammals, fungi, and insects. Two CK2 regulatory subunits, CKB3 and CKB4, have been also linked with the Arabidopsis thaliana circadian system. However, the biological relevance and the precise mechanisms of CK2 function within the plant clockwork are not known. By using ChIP and Double-ChIP experiments together with in vivo luminescence assays at different temperatures, we were able to identify a temperature-dependent function for CK2 modulating circadian period length. Our study uncovers a previously unpredicted mechanism for CK2 antagonizing the key clock regulator CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1). CK2 activity does not alter protein accumulation or subcellular localization but interferes with CCA1 binding affinity to the promoters of the oscillator genes. High temperatures enhance the CCA1 binding activity, which is precisely counterbalanced by the CK2 opposing function. Altering this balance by over-expression, mutation, or pharmacological inhibition affects the temperature compensation profile, providing a mechanism by which plants regulate circadian period at changing temperatures. Therefore, our study establishes a new model demonstrating that two opposing and temperature-dependent activities (CCA1-CK2) are essential for clock temperature compensation in Arabidopsis.


Developmental signature, synaptic connectivity and neurotransmission are conserved between vertebrate hair cells and tunicate coronal cells.
In tunicates, the coronal organ represents a sentinel checking particle entrance into the pharynx. The organ differentiates from an anterior embryonic area considered a proto-placode. For their embryonic origin, morphological features and function, coronal sensory cells have been hypothesized to be homologues to vertebrate hair cells. However, vertebrate hair cells derive from a posterior placode. This contradicts one of the principle historical criteria for homology, similarity of position, which could be taken as evidence against coronal cells/hair cells homology. In the tunicates Ciona intestinalis and C. robusta, we found that the coronal organ expresses genes (Atoh, Notch, Delta-like, Hairy-b, and Musashi) characterizing vertebrate neural and hair cell development. Moreover, coronal cells exhibit a complex synaptic connectivity pattern, and express neurotransmitters (Glu, ACh, GABA, 5-HT, and catecholamines), or enzymes for their synthetic machinery, involved in hair cell activity. Lastly, coronal cells express the Trpa gene, which encodes an ion channel expressed in hair cells. These data lead us to hypothesize a model in which competence to make secondary mechanoreceptors was initially broadly distributed through placode territories, but has become confined to different placodes during the evolution of the vertebrate and tunicate lineages.


Structural Revision of Baulamycin A and Structure-Activity Relationships of Baulamycin A Derivatives.
Total synthesis of the proposed structure of baulamycin A was performed. The spectral properties of the synthetic compound differ from those reported for the natural product. On the basis of comprehensive NMR study, we proposed two other possible structures for natural baulamycin A. Total syntheses of these two substances were performed, which enabled assignment of the correct structure of baulamycin A. Key features of the convergent and fully stereocontrolled route include Evans Aldol and Brown allylation reactions to construct the left fragment, a prolinol amide-derived alkylation/desymmetrization to install the methyl-substituted centers in the right fragment, and finally, a Carreira alkynylation to join both fragments. In addition, we have determined the inhibitory activities of novel baulamycin A derivatives against the enzyme SbnE. This SAR study provides useful insight into the design of novel SbnE inhibitors that overcome the drug resistance of pathogens, which cause life-threatening infections.


Public attitudes towards smoking and tobacco control policy in Russia.
Since the political transition in 1991, Russia has been targeted intensively by the transnational tobacco industry. Already high smoking rates among men have increased further; traditionally low rates among women have more than doubled. The tobacco companies have so far faced little opposition as they shape the discourse on smoking in Russia. This paper asks what ordinary Russians really think about possible actions to reduce smoking. A representative sample of the Russian population (1600 respondents) was interviewed face to face in November 2007. Only 14% of respondents considered tobacco control in Russia adequate, while 37% thought that nothing was being done at all. There was support for prices keeping pace with or even exceeding inflation. Over 70% of all respondents favoured a ban on sales from street kiosks, while 56% believed that existing health warnings (currently 4% of front and back of packs) were inadequate. The current policy of designating a few tables in bars and restaurants as non-smoking was supported by less than 10% of respondents, while almost a third supported a total ban, with 44% supporting provision of equal space for smokers and non-smokers. Older age, non-smoking status and living in a smaller town all emerged as significantly associated with the propensity to support antismoking measures. The tobacco companies were generally viewed as behaving like most other companies in Russia, with three-quarters of respondents believing that these companies definitely or maybe bribe politicians. Knowledge of impact of smoking on health was limited with significant underestimation of dangers and addictive qualities of tobacco. A third believed that light cigarettes are safer than normal cigarettes. The majority of the Russian population would support considerable strengthening of tobacco control policies but there is also a need for effective public education campaigns.


Mindfulness meditation improves cognition: evidence of brief mental training.
Although research has found that long-term mindfulness meditation practice promotes executive functioning and the ability to sustain attention, the effects of brief mindfulness meditation training have not been fully explored. We examined whether brief meditation training affects cognition and mood when compared to an active control group. After four sessions of either meditation training or listening to a recorded book, participants with no prior meditation experience were assessed with measures of mood, verbal fluency, visual coding, and working memory. Both interventions were effective at improving mood but only brief meditation training reduced fatigue, anxiety, and increased mindfulness. Moreover, brief mindfulness training significantly improved visuo-spatial processing, working memory, and executive functioning. Our findings suggest that 4days of meditation training can enhance the ability to sustain attention; benefits that have previously been reported with long-term meditators.


How Habitat Edges Change Species Interactions.
Traditionally, ecologists interested in habitat edges have focused on edge-related gradients in patterns of species richness or abiotic variables. Here, however, we take a different perspective, attempting to synthesize recent empirical results concerning the effects of habitat edges on population dynamics with contemporary theoretical developments to outline the ways in which species interactions, and the dynamics of the communities in which they are embedded, can be changed by habitat edges. We find a striking convergence between empirical notions of a patch's core area and analytical results from partial differential equation models. A review of both empirical and theoretical studies suggests four general classes of mechanisms through which habitat edges can have similar impacts on dissimilar types of species interactions. Specifically, we focus on edges' roles as dispersal barriers or filters, edges' influences on mortality, edges' involvement in spatial subsidies (in which dispersers' intrapatch impacts are maintained by their activities in other habitats), and edges' roles as generators of novel interactions. For each class of edge-mediated effects, we provide examples of how one can use spatial modeling to address the relevant questions on these topics, which together form a key link between community dynamics and landscape structure.


Self-Organization in Pattern Formation.
Self-organization is pervasive in development, from symmetry breaking in the early embryo to tissue patterning and morphogenesis. For a few model systems, the underlying molecular and cellular processes are now sufficiently characterized that mathematical models can be confronted with experiments, to explore the dynamics of pattern formation. Here, we review selected systems, ranging from cyanobacteria to mammals, where different forms of cell-cell communication, acting alone or together with positional cues, drive the patterning of cell fates, highlighting the insights that even very simple models can provide as well as the challenges on the path to a predictive understanding of development.


Mortality differences among organisms causing septicemia in hemodialysis patients.
Septicemia is a serious problem in hemodialysis patients because it can lead to life-threatening complications and a persistently elevated risk of death. Most analyses have not examined whether there are differences in mortality risk among the organisms that cause these episodes of septicemia. This study was a retrospective cohort analysis of first septicemia hospitalizations during the first year of hemodialysis. Time to death (both in-hospital and within 12 weeks post-discharge) was compared among the different septicemia-causing organisms based on discharge diagnoses in Medicare billing data from 1996 to 2001. The effect of various complications on mortality risk was also evaluated. There were 22,130 septicemia hospitalizations identified. The most common organism identified was Staphylococcus aureus (27%), with no other organism having an incidence >10%. The overall unadjusted death rate from admission through 12 weeks of follow-up was 34%. During the first hospitalization, the death rate was 14%, and during the 12-week period after the hospitalization it was 20%. In adjusted analyses, S. aureus was associated with a 20% higher risk of death both during the in-hospital period and the 12-week post-discharge period, when compared with all other specified organisms. Hospitalizations complicated by meningitis, stroke, or endocarditis were also associated with increased risk of mortality, independent of the organism causing septicemia. Septicemia hospitalizations are associated with a high mortality rate--both during the initial hospitalization and after discharge. Meningitis, stroke, and endocarditis represent particularly serious complications. Overall, septicemia hospitalizations (especially for S. aureus) are serious events, and patients would benefit from better treatment and prevention.


Repeatability of Measuring Corneal Nerve Migration Rate in Individuals With and Without Diabetes.
To assess the repeatability of measuring the corneal nerve migration rate in individuals with and without neuropathy. Wide-field montages of the subbasal corneal nerve plexus were generated at baseline and after 3 weeks for 14 participants. Montages were manually examined side by side to identify a referent landmark in the inferior whorl region (origin) and throughout each montage. A software program was developed to measure nerve migration of all identified points relative to the origin. Repeatability was determined by measurement of nerve migration for within observer (one researcher on 2 occasions, 5 days apart) and between observers (2 observers) within 4 different zones based on the distance from the origin and in the vertical section of the wide-field montage. The impact of images being montaged with fully automated software on repeatability was also investigated. The mean difference between observations 1 and 2 for observer 1 was 0.02 ± 1.3 μm/wk (P = 0.94), with an intraclass correlation coefficient (ICC) of 0.99 [95% confidence interval (CI) = 0.99-1.00], and the mean difference between observer 1 and 2 was 0.3 ± 1.2 μm/wk (P = 0.41), with an ICC of 0.99 (95% CI = 0.99-1.00). The mean difference between observations 1 (images montaged by semiautomated software) and 2 (images montaged by fully automated software) was 1.2 ± 4.9 μm/wk (P = 0.41), with an ICC of 0.96 (95% CI = 0.87-1.00). Measuring corneal nerve migration rate is highly repeatable for within and between observers and when using different methods of image montaging.


Ultrasonically actuated silicon microprobes for cardiac signal recording.
In this paper, we report on ultrasonically actuated silicon thin microprobes that successfully penetrated canine cardiac tissue in vitro, and recorded the electrophysiological signals from multiple sites simultaneously within the heart wall. The penetration force--maximum force encountered by the probe during penetration--is found to reduce with increasing ultrasonic driving voltage, on both excised canine right ventricular muscle and chicken breast muscle. The rate of force decrease varies with tissue type and microprobe dimension. With ultrasonic actuation, the silicon microprobes are inserted into isolated perfused canine heart without breakage or significant buckling, under 10Vpp actuating voltage. Recordings were obtained from isolated perfused canine heart during pacing, following the induction of ventricular tachycardia, and during the transition from ventricular tachycardia to ventricular fibrillation. Local conduction velocity of 0.60 +/- 0.03 m/s was observed from the multichannel recordings from the canine right ventricular wall under epicardial pacing. The application of the ultrasonic microprobes in cardiac electrophysiology study can provide information for reconstruction of electrical wave propagation within the heart, which is important to understanding the mechanisms of cardiac arrhythmias.


Polymersomes: Breaking the Glass Ceiling?
Polymer vesicles, also known as polymersomes, have garnered a lot of interest even before the first report of their fabrication in the mid-1990s. These capsules have found applications in areas such as drug delivery, diagnostics and cellular models, and are made via the self-assembly of amphiphilic block copolymers, predominantly with soft, rubbery hydrophobic segments. Comparatively, and despite their remarkable impermeability, glassy polymersomes (GPs) have been less pervasive due to their rigidity, lack of biodegradability and more restricted fabrication strategies. GPs are now becoming more prominent, thanks to their ability to undergo stable shape-change (e.g., into non-spherical morphologies) as a response to a predetermined trigger (e.g., light, solvent). The basics of block copolymer self-assembly with an emphasis on polymersomes and GPs in particular are reviewed here. The principles and advantages of shape transformation of GPs as well as their general usefulness are also discussed, together with some of the challenges and opportunities currently facing this area.


Functional subclasses of T lymphocytes bearing different Ly antigens. II. Cooperation between subclasses of Ly+ cells in the generation of killer activity.
Lymphocytes from BALB/c or C57B6/6 mice that develop killer activity to alloantigens belong to the numerically small Ly-23 subclass of peripheral T cells, distinguished by selective expression of Ly-23 determinants on their surfaces. The maturation of these cells to killer cells can be amplified by Ly-1+ cells, which do not themselves contribute to the killer cell pool. This amplification was abolished by escluding Ia("Beta")+ cells from the stimulator population during mixed lymphocyte culture (MCL), suggesting that amplification is due to selective recognition of I region antigens by L-1+ cells, a conclusion already drawn from our previous evidence that Ia differences activate Ly-1+ cells but not Ly-23+ cells. These and other experiments indicate that amplification of killer cell production in vitro by Ly-1+ cells is not due to their conversion to Ly-23+ cells during MLC, but to their ability to recognize major histocompatibility complex determinants not recognized by Ly-23+ cells.


Zinc protoporphyrin inhibition of lipopolysaccharide-, lipoteichoic acid-, and peptidoglycan-induced nitric oxide production through stimulating iNOS protein ubiquitination.
In the present study, zinc protoporphyrin (ZnPP), but not ferric protoporphyrin (FePP), tin protoporphyrin (SnPP), or zinc chloride (ZnCl(2)), at the doses of 0.5, 1, and 2 microM, dose-dependently inhibited lipopolysaccharide- (LPS), lipoteichoic acid (LTA), and peptidoglycan (PGN)-induced inducible nitric oxide (iNOS) and nitric oxide (NO) production with an increase in heme oxygenase 1 (HO-1) protein in RAW264.7 macrophages in a serum-free condition. NO inhibition and HO-1 induction by ZnPP were blocked by the separate addition of fetal bovine serum (FBS) and bovine serum albumin (BSA). A decrease in the iNOS/NO ratio and an increase in HO-1 protein by ZnPP were identified in three different conditions including ZnPP pretreatment, ZnPP co-treatment, and ZnPP post-treatment with LPS and LTA. Activation of c-Jun N-terminal kinases (JNKs) and extracellular regulated kinases (ERKs) were detected in LPS-, LTA-, and PGN-treated RAW264.7 cells, and iNOS/NO production was blocked by adding the JNK inhibitor, SP600125, but not the ERK inhibitor, PD98059. However, ZnPP addition potentiated ERK and JNK protein phosphorylation stimulated by LPS, LTA, and PGN. Increases in total protein ubiquitination and ubiquitinated iNOS proteins were detected in ZnPP-treated macrophages elicited by LPS according to Western and immunoprecipitation/Western blotting assays, respectively. The decrease in LPS-induced iNOS protein by ZnPP was reversed by adding the proteasome inhibitors MG132 and lactacystin. The reduction in HO-1 protein induced by ZnPP via transfection of HO-1 small interfering RNA did not affect the inhibitory effect of ZnPP against LPS-induced iNOS/NO production and protein ubiquitination induced by ZnPP in macrophages. Data of the present study provide the first evidence to support ZnPP effectively inhibiting inflammatory iNOS/NO production through activation of protein ubiquitination in a HO-1-independent manner in macrophages.


Trimethoprim resistance in Haemophilus influenzae is due to altered dihydrofolate reductase(s).
We characterized a highly purified preparation of the chromosomally encoded dihydrofolate reductase (DHFR) from a trimethoprim-susceptible (Tmp8; strain MAP) and two trimethoprim-resistant (TmpR) strains (MAP/47 and MAP/42) of Haemophilus influenzae. The enzymes were purified between 650- and 3000-fold by gel-filtration and dye-ligand chromatography. The apparent molecular mass of the three proteins was 18400 Da by PAGE under denaturing and nondenaturing conditions. Total enzyme activity was greater in all fractions from the TmpR strains compared with the Tmp8 isolate. The three enzymes had a similar Km for dihydrofolate (7, 9 and 5 microM) and NADPH (2, 5 and 6 microM). However, the Tmp IC50 (the concentration necessary for 50% inhibition of DHFR activity) for the Tmp8 strain MAP was 0.001 microM, whereas DHFR from the TmpR strains MAP/47 and MAP/42 had values of 0.1 microM and 0.3 microM respectively. The methotrexate IC50 of the MAP/42 DHFR was 0.06 microM in comparison with the enzyme from MAP (0.008 microM) and MAP/47 (0.007 microM). Isoelectric focusing indicated that the DHFR from MAP/42 had a different isoelectric point (pI 7.6) compared with the enzymes from MAP and MAP/47 (pI 7.3). Peptide mapping after digestion with trypsin revealed one major peptide fragment (7.9 kDa) in the DHFR of MAP and MAP/47 and three major tryptic fragments (7.9, 9.6 and 12.5 kDa) in DHFR from MAP/42. We conclude that trimethoprim resistance in H. influenzae results from overproduction of structurally altered DHFR(s).


Speciality preferences in Dutch medical students influenced by their anticipation on family responsibilities.
Physician gender is associated with differences in the male-to-female ratio between specialities and with preferred working hours. We explored how graduating students' sex or full-time or part-time preference influences their speciality choice, taking work-life issues into account. Graduating medical students at Radboud University Medical Centre, the Netherlands participated in a survey (2008-2012) on career considerations. Logistic regression tested the influence of sex or working hour preference on speciality choice and whether work-life issues mediate. Of the responding students (N = 1,050, response rate 83, 73.3 % women), men preferred full-time work, whereas women equally opted for part time. More men chose surgery, more women family medicine. A full-time preference was associated with a preference for surgery, internal medicine and neurology, a part-time preference with psychiatry and family medicine. Both male and female students anticipated that foremost the career of women will be negatively influenced by family life. A full-time preference was associated with an expectation of equality in career opportunities or with a less ambitious partner whose career would affect family life. This increased the likelihood of a choice for surgery and reduced the preference for family medicine among female students. Gender specifically plays an important role in female graduates' speciality choice making, through considerations on career prospects and family responsibilities.


Cognitive deficits in Machado-Joseph disease correlate with hypoperfusion of visual system areas.
Cognitive and olfactory impairments have previously been demonstrated in patients with spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph disease (MJD)-SCA3/MJD. We investigated changes in regional cerebral blood flow (rCBF) using single-photon emission computed tomography (SPECT) imaging in a cohort of Brazilian patients with SCA3/MJD. The aim of the present study was to evaluate the correlation among rCBF, cognitive deficits, and olfactory dysfunction in SCA3/MJD. Twenty-nine genetically confirmed SCA3/MJD patients and 25 control subjects were enrolled in the study. The severity of cerebellar symptoms was measured using the International Cooperative Ataxia Rating Scale and the Scale for the Assessment and Rating of Ataxia. Psychiatric symptoms were evaluated by the Hamilton Anxiety Scale and Beck Depression Inventory. The neuropsychological assessment consisted of Spatial Span, Symbol Search, Picture Completion, the Stroop Color Word Test, Trail Making Test (TMT), and Phonemic Verbal Fluency. Subjects were also submitted to odor identification evaluation using the 16-item Sniffin' Sticks. SPECT was performed using ethyl cysteine dimer labeled with technetium-99m. SCA3/MJD patients showed reduced brain perfusion in the cerebellum, temporal, limbic, and occipital lobes compared to control subjects (pFDR <0.001). A significant positive correlation was found between the Picture Completion test and perfusion of the left parahippocampal gyrus and basal ganglia in the patient group as well as a negative correlation between the TMT part A and bilateral thalamus perfusion. The visuospatial system is affected in patients with SCA3/MJD and may be responsible for the cognitive deficits seen in this disease.


A haemagglutination test for rapid detection of antibodies to SARS-CoV-2.
Serological detection of antibodies to SARS-CoV-2 is essential for establishing rates of seroconversion in populations, and for seeking evidence for a level of antibody that may be protective against COVID-19 disease. Several high-performance commercial tests have been described, but these require centralised laboratory facilities that are comparatively expensive, and therefore not available universally. Red cell agglutination tests do not require special equipment, are read by eye, have short development times, low cost and can be applied at the Point of Care. Here we describe a quantitative Haemagglutination test (HAT) for the detection of antibodies to the receptor binding domain of the SARS-CoV-2 spike protein. The HAT has a sensitivity of 90% and specificity of 99% for detection of antibodies after a PCR diagnosed infection. We will supply aliquots of the test reagent sufficient for ten thousand test wells free of charge to qualified research groups anywhere in the world.


[Association between body mass index and coronary heart disease in Qingdao: a prospective study].
Objective: To analyze the association between body mass index (BMI) and coronary heart disease. Methods: The data for the present study were from the prospective cohort study of China Kadoorie Biobank (CKB) in Qingdao, a total of 33 355 participants aged 30-79 years were included in the study. Cox regression analyses were performed to evaluate the association between BMI and coronary heart disease. Results: During the follow-up for an average 9.2 years, a total of 2 712 cases of ischemic heart disease (IHD) and 420 cases of major coronary events (MCE) were found. Multivariate Cox regression analysis showed that, compared with participants with normal BMI, the participants who were overweight had a 41% and 87% higher risk of IHD and MCE, the adjusted HR were 1.41 (95%CI: 1.27-1.56) and 1.87 (95%CI: 1.43-2.44), respectively. The participants who were obesity had 91% and 143% higher risk of IHD and MCE, the adjusted HR were 1.91 (95%CI: 1.72-2.13) and 2.43 (95%CI: 1.82-3.24), respectively. Conclusion: Overweight and obesity might increase the risk for IHD and MCE.


From Data to Action: Neuroepidemiology Informs Implementation Research for Global Stroke Prevention and Treatment.
As a scientific field of study, neuroepidemiology encompasses more than just the descriptive study of the frequency, distribution, determinants and outcomes of neurologic diseases in populations. It also includes experimental aspects that span the full spectrum of clinical and population science research. As such, neuroepidemiology has a strong potential to inform implementation research for global stroke prevention and treatment. This review begins with an overview of the progress that has been made in descriptive and experimental neuroepidemiology over the past quarter century with emphasis on standards for evidence generation, critical appraisal of that evidence and impact on clinical and public health practice at the national, regional and global levels. Specific advances made in high-income countries as well as in low- and middle-income countries are presented. Gaps in implementation as well as evidence gaps in stroke research, stroke burden, clinical outcomes and disparities between developed and developing countries are then described. The continuing need for high quality neuroepidemiologic data in low- and middle-income countries is highlighted. Additionally, persisting disparities in stroke burden and care by sex, race, ethnicity, income and socioeconomic status are discussed. The crucial role that national stroke registries have played in neuroepidemiologic research is also addressed. Opportunities presented by new directions in comparative effectiveness and implementation research are discussed as avenues for turning neuroepidemiological insights into action to maximize health impact and to guide further biomedical research on neurological diseases.


[Opinion of primary care physicians from Ourense on various features of the pharmaceutical prescription].
To learn the opinion of the primary care Physicians of Ourense (Spain) with respect to certain aspects of their prescription of medicines, such as their awareness of the price of drugs, the induced prescribing perceived, their relationship with the pharmaceutical industry and their opinions concerning possible measures for reducing the expenditure on medicines. In this transversal descriptive study, all of the primary care physicians in the province of Ourense (243) were surveyed by means of a previously-approved questionnaire sent to them by post. The questionnaire included demographic characteristics of the physicians, the influence of cost when prescribing medicines, their estimate of the price of 15 drugs routinely prescribed and their opinion on different aspects relating to induced prescription, the pharmaceutical industry and different strategies in order to curtail the expenditure on medicines. In order to analyze the results, the chi 2 and Student's t tests and an analysis of the variance were used, together with the Spearman correlation coefficient with alpha = 0.05. The level of participation of the physicians surveyed was 42.8% (104). The average daily duration of visits by pharmaceutical company representatives was 13.6 minutes, with new specialities being the topic of greatest interest. Close to 27% admitted to having participated in clinical testing sponsored by the industry. 23% considered the price to be a priority when prescribing. Induced prescription accounted for 39.7% of the total. The majority of physicians chose co-payment as the means for reducing expenditure on medicines. In the estimate of prices, the overall percentage of error was 45.7%, underestimating the more expensive medicines and overestimating the less expensive. There is a significant lack of awareness of the price of drugs among the primary care physicians. Most of the physicians do not feel that the price of a drug should be a priority when prescribing medicines. There is a high percentage of induced prescription perceived. With regard to the expenditure on pharmaceuticals, the majority of those surveyed feel that financial measures, specifically co-payment, are the most effective in order to curtail rising costs.


Comparison of a label-free quantitative proteomic method based on peptide ion current area to the isotope coded affinity tag method.
Recently, several research groups have published methods for the determination of proteomic expression profiling by mass spectrometry without the use of exogenously added stable isotopes or stable isotope dilution theory. These so-called label-free, methods have the advantage of allowing data on each sample to be acquired independently from all other samples to which they can later be compared in silico for the purpose of measuring changes in protein expression between various biological states. We developed label free software based on direct measurement of peptide ion current area (PICA) and compared it to two other methods, a simpler label free method known as spectral counting and the isotope coded affinity tag (ICAT) method. Data analysis by these methods of a standard mixture containing proteins of known, but varying, concentrations showed that they performed similarly with a mean squared error of 0.09. Additionally, complex bacterial protein mixtures spiked with known concentrations of standard proteins were analyzed using the PICA label-free method. These results indicated that the PICA method detected all levels of standard spiked proteins at the 90% confidence level in this complex biological sample. This finding confirms that label-free methods, based on direct measurement of the area under a single ion current trace, performed as well as the standard ICAT method. Given the fact that the label-free methods provide ease in experimental design well beyond pair-wise comparison, label-free methods such as our PICA method are well suited for proteomic expression profiling of large numbers of samples as is needed in clinical analysis.


Thiazolidinedione ameliorates renal injury in experimental diabetic rats through anti-inflammatory effects mediated by inhibition of NF-kappaB activation.
Thiazolidinedione (TZD), a ligand for peroxisome proliferator-activated receptor-gamma (PPAR-gamma), exerts anti-inflammatory effects independently of the insulin-sensitizing effect. In the present study, we tested the hypothesis that TZD prevents the progression of diabetic nephropathy by modulating the inflammatory process. Five-week-old Sprague-Dawley rats were divided into three groups: 1) nondiabetic control rats (non-DM), 2) diabetic rats (DM), and 3) diabetic rats treated with pioglitazone (DM+pio). Diabetes was induced by injection with streptozotocin (STZ). The DM+pio group received 0.0002% pioglitazone mixed in chow for 8 wk after induction of diabetes. Blood glucose and HbA1c were elevated in diabetic rats but did not change by treatment with pioglitazone. Pioglitazone reduced urinary albumin excretion and glomerular hypertrophy, suppressed the expression of transforming growth factor (TGF)-beta, type IV collagen, and ICAM-1, and infiltration of macrophages in the kidneys of diabetic rats. Furthermore, renal NF-kappaB activity was increased in diabetic rats and reduced by pioglitazone. PPAR-gamma was expressed in glomerular endothelial cells in the diabetic kidney and in cultured glomerular endothelial cells. High-glucose conditions increased the expression of ICAM-1 and the activation of NF-kappaB in cultured glomerular endothelial cells. These changes were reduced by pioglitazone, ciglitazone, and pyrrolidine dithiocarbamate, an inhibitor of NF-kappaB. However, pioglitazone did not show the changes in the presence of PPAR-gamma antagonist GW9662. Our results suggest that the preventive effects of pioglitazone may be mediated by its anti-inflammatory actions, including inhibition of NF-kappaB activation, ICAM-1 expression, and macrophage infiltration in the diabetic kidney.


Adverse events in apheresis: An update of the WAA registry data.
Apheresis with different procedures and devices are used for a variety of indications that may have different adverse events (AEs). The aim of this study was to clarify the extent and possible reasons of various side effects based on data from a multinational registry. The WAA-apheresis registry data focus on adverse events in a total of 50846 procedures in 7142 patients (42% women). AEs were graded as mild, moderate (need for medication), severe (interruption due to the AE) or death (due to AE). More AEs occurred during the first procedures versus subsequent (8.4 and 5.5%, respectively). AEs were mild in 2.4% (due to access 54%, device 7%, hypotension 15%, tingling 8%), moderate in 3% (tingling 58%, urticaria 15%, hypotension 10%, nausea 3%), and severe in 0.4% of procedures (syncope/hypotension 32%, urticaria 17%, chills/fever 8%, arrhythmia/asystole 4.5%, nausea/vomiting 4%). Hypotension was most common if albumin was used as the replacement fluid, and urticaria when plasma was used. Arrhythmia occurred to similar extents when using plasma or albumin as replacement. In 64% of procedures with bronchospasm, plasma was part of the replacement fluid used. Severe AEs are rare. Although most reactions are mild and moderate, several side effects may be critical for the patient. We present side effects in relation to the procedures and suggest that safety is increased by regular vital sign measurements, cardiac monitoring and by having emergency equipment nearby.


A polymorphism in the 3' untranslated region of the gene encoding prostaglandin endoperoxide synthase 2 is not associated with an increase in breast cancer risk: a nested case-control study.
Prostaglandins are integral components in the cellular response to inflammation, promoting cellular proliferation and angiogenesis. The enzyme responsible for the conversion of arachidonic acid to prostaglandins in response to inflammation is prostaglandin endoperoxide synthase 2/cyclo-oxygenase 2 (PTGS2/COX2). Polymorphisms in the PTGS2 gene have been associated with various diseases, including inflammatory bowel disease and cancer of the lung, colorectum, and breast. We genotyped the five most common polymorphisms (rs20417, rs5277, rs20432, rs5275, and rs4648298) in the Nurses' Health Study (1,270 cases, 1,762 controls) to test the hypothesis that polymorphisms in PTGS2 are associated with breast cancer risk, using logistic regression analyses. The Nurses' Health Study 2 (317 cases, 634 controls) and Harvard Women's Health Study (702 cases, 703 controls) were used to further examine putative associations. The rs5275 polymorphism in the 3' untranslated region of the PTGS2 gene was associated with a decrease in breast cancer risk. We therefore genotyped this single-nucleotide polymorphism in the Nurses' Health Study 2 and Harvard Women's Health Study. Similar results were observed in these subsequent analyses, with no statistically significant heterogeneity in risk estimates between studies. In pooled analyses, women homozygous for the T allele at rs5275 had a 20% lower risk of breast cancer than those homozygous for the C allele (odds ratio 0.80, 95% confidence interval 0.66 to 0.97). Although this polymorphism may be associated with a decrease in breast cancer risk among Caucasian women, we provide strong evidence that it is not associated with an increased risk of breast cancer.


Determining elasticity from single polymer dynamics.
The ability to determine polymer elasticity and force-extension relations from polymer dynamics in flow has been challenging, mainly due to difficulties in relating equilibrium properties such as free energy to far-from-equilibrium processes. In this work, we determine polymer elasticity from the dynamic properties of polymer chains in fluid flow using recent advances in statistical mechanics. In this way, we obtain the force-extension relation for DNA from single molecule measurements of polymer dynamics in flow without the need for optical tweezers or bead tethers. We further employ simulations to demonstrate the practicality and applicability of this approach to the dynamics of complex fluids. We investigate the effects of flow type on this analysis method, and we develop scaling laws to relate the work relation to bulk polymer viscometric functions. Taken together, our results show that nonequilibrium work relations can play a key role in the analysis of soft material dynamics.


Multimodality imaging for characterization, classification, and staging of malignant pleural mesothelioma.
Malignant pleural mesothelioma (MPM) is the most common primary malignancy of the pleura and is associated with asbestos exposure in approximately 80% of patients. The patient prognosis is poor, with a median survival of 9-17 months after diagnosis. However, improved survival and decreased morbidity and mortality have been demonstrated when the diagnosis is made in the early stages of disease and specific treatment strategies are implemented. A staging system that focuses on the extent of primary tumor (T), lymph node involvement (N), and metastatic disease (M) has been devised by the International Mesothelioma Interest Group and emphasizes factors related to overall survival. Radiologists should recognize the manifestations of MPM across multiple imaging modalities, translate these findings into the updated staging system, and understand the effects of appropriate staging on treatment and survival. Computed tomography (CT) remains the primary imaging modality used to evaluate MPM and efficiently demonstrates the extent of primary tumor, intrathoracic lymphadenopathy, and extrathoracic spread. However, additional imaging modalities, such as magnetic resonance (MR) imaging of the thorax and positron emission tomography (PET)/CT with fluorodeoxyglucose, have emerged in recent years and are complementary to CT for disease staging and evaluation of patients with MPM. Thoracic MR imaging is particularly useful for identifying invasion of the chest wall, mediastinum, and diaphragm, and PET/CT can accurately demonstrate intrathoracic and extrathoracic lymphadenopathy and metastatic disease.


A joint Bayesian approach for the analysis of response measured at a primary endpoint and longitudinal measurements.
Joint mixed modeling is an attractive approach for the analysis of a scalar response measured at a primary endpoint and longitudinal measurements on a covariate. In the standard Bayesian analysis of these models, measurement error variance and the variance/covariance of random effects are a priori modeled independently. The key point is that these variances cannot be assumed independent given the total variation in a response. This article presents a joint Bayesian analysis in which these variance terms are a priori modeled jointly. Simulations illustrate that analysis with multivariate variance prior in general lead to reduced bias (smaller relative bias) and improved efficiency (smaller interquartile range) in the posterior inference compared with the analysis with independent variance priors.


Coinfection with Bartonella clarridgeiae and Bartonella henselae and with different Bartonella henselae strains in domestic cats.
Bartonella clarridgeiae and several strains of Bartonella henselae, the agent of cat scratch disease, with variations in the 16S rRNA gene have been found to infect the blood of cats. An epidemiologic study of Bartonella infection in domestic French cats revealed that of 436 cats sampled, 5 cats (1.1%) were coinfected with B. henselae and B. clarridgeiae and 2 cats (0.5%) were coinfected with two strains of B. henselae with variations in the 16S rRNA gene, B. henselae type I and type II. In an indirect immunofluorescence assay, coinfected cats tested positive for both Bartonella species at titers of > or = 128. Identification of the colonies was achieved by preformed enzyme analysis, PCR-restriction fragment length polymorphism analysis of the citrate synthase gene, and 16S rRNA gene sequencing. Colony size differences in mixed culture allowed differentiation of the Bartonella species. The coinfection of cats with two Bartonella species or variants of the same species raises concern about the possibility of dual infection in humans. The development of a polyvalent vaccine targeted against the most pathogenic or invasive strains may be a means of protecting cats and man from infection.


Non-invasive positive pressure ventilation for the treatment of severe stable chronic obstructive pulmonary disease: a prospective, multicentre, randomised, controlled clinical trial.
Evidence is weak for the ability of long-term non-invasive positive pressure ventilation (NPPV) to improve survival in patients with stable hypercapnic chronic obstructive pulmonary disease (COPD). Previous prospective studies did not target a reduction in hypercapnia when adjusting ventilator settings. This study investigated the effect of long-term NPPV, targeted to markedly reduce hypercapnia, on survival in patients with advanced, stable hypercapnic COPD. This investigator-initiated, prospective, multicentre, randomised, controlled clinical trial enrolled patients with stable GOLD stage IV COPD and a partial carbon dioxide pressure (PaCO2) of 7 kPa (51.9 mm Hg) or higher and pH higher than 7.35. NPPV was targeted to reduce baseline PaCO2 by at least 20% or to achieve PaCO2 values lower than 6.5 kPa (48.1 mm Hg). Patients were randomly assigned (in a 1:1 ratio) via a computer-generated randomisation sequence with a block size of four, to continue optimised standard treatment (control group) or to receive additional NPPV for at least 12 months (intervention group). The primary outcome was 1-year all-cause mortality. Analysis was by intention to treat. The intervention was unblinded, but outcome assessment was blinded to treatment assignment. This study is registered with ClinicalTrials.gov, number NCT00710541. Patients were recruited from 36 respiratory units in Germany and Austria, starting on Oct 29, 2004, and terminated with a record of the vital status on July 31, 2011. 195 patients were randomly assigned to the NPPV group (n=102) or to the control group (n=93). All patients from the control group and the NPPV group were included in the primary analysis. 1-year mortality was 12% (12 of 102 patients) in the intervention group and 33% (31 of 93 patients) in the control group; hazard ratio 0.24 (95% CI 0.11-0.49; p=0.0004). 14 (14%) patients reported facial skin rash, which could be managed by changing the type of the mask. No other intervention-related adverse events were reported. The addition of long-term NPPV to standard treatment improves survival of patients with hypercapnic, stable COPD when NPPV is targeted to greatly reduce hypercapnia. German Lung Foundation; ResMed, Germany; Tyco Healthcare, Germany; and Weinmann, Germany.


Identification of pathogenesis-associated genes by T-DNA-mediated insertional mutagenesis in Botrytis cinerea: a type 2A phosphoprotein phosphatase and an SPT3 transcription factor have significant impact on virulence.
Agrobacterium tumefaciens-mediated transformation (ATMT) was used to generate an insertional mutant library of the gray mold fungus Botrytis cinerea. From a total of 2,367 transformants, 68 mutants showing significant reduction in virulence on tomato and bean plants were analyzed in detail. As reported for other fungal ATMT libraries, integrations were mostly single copy, occurred preferentially in noncoding (regulatory) regions, and were frequently accompanied by small deletions of the target sequences and loss of parts of the border sequence. Two T-DNA integration events that were found to be linked to virulence were characterized in more detail: a catalytic subunit of a PP2A serine/threonine protein phosphatase (BcPP2Ac) and the SPT3 subunit of a Spt-Ada-Gcn5-acetyltransferase (SAGA-like) transcriptional regulator complex. Gene replacement and silencing approaches revealed that both Bcpp2Ac and SPT3 are crucial for virulence, growth, and differentiation as well as for resistance to H(2)O(2) in B. cinerea.


Synchrotron X-ray diffraction and histochemical studies of normal and myopic chick eyes.
Synchrotron X-ray diffraction patterns were obtained from the cornea and posterior sclera of control and myopic chicks. No significant differences was found in the interfibrillar or in the intermolecular spacing of the collagen fibrils from the corneas of control and myopic chicks. The intermolecular spacing of myopic sclera was shown to be significantly (p < 0.01) higher than in controls. Sclera and cornea from normal and myopic chicks were stained for proteoglycans using the 'critical electrolyte' method of Scott and Orford (1981). In the sclera, two morphologically distinct types of proteoglycans could be distinguished; one small and usually elongated (approximately 20 nm), the other larger and irregularly shaped. The small proteoglycans were seen binding preferentially to the 'd' and 'e' bands of the collagen fibrils. Small proteoglycans were also present within the fibrils, these were usually approximately 10 nm in diameter although sizes up to 30 nm were observed. Longitudinal sections of fibrils revealed that these intrafibrillar proteoglycans were chiefly orientated parallel to the axis of the collagen fibrils, and preferentially located along the gap region of the fibrils. No difference was observed in the binding sites of the proteoglycans between normal and myopic sclera. The larger proteoglycans were also seen aggregated into clumps, which were usually associated with spaces between collagen bundles. The differences between control and myopic sclera appear to be quantitative rather than qualitative suggesting that the scleral expansion in myopia is an enhanced form of normal scleral growth.


ATP-dependent regulation of a G protein-coupled K+ channel (GIRK1/GIRK4) expressed in oocytes.
Recent studies suggest that activation of the atrial muscarinic K+ current by acetylcholine (ACh) involves an ATP-dependent process that is then inhibited by a cytosolic protein to result in the rapid desensitization. To obtain further evidence in support of such a dually regulated process, we studied the properties of GIRK1 and GIRK4, which, when coexpressed in oocytes, form a heteromultimer that closely resembles the muscarinic K+ channel. ACh activated an inwardly rectifying K+ current that desensitized slowly. In cell-attached patches with ACh in the pipette, the mean open times (tau zero) of GIRK1/GIRK4 were 1.2 +/- 0.1 (28%) and 6.7 +/- 0.8 ms (72%) and did not change significantly with time. However, in inside-out patches, the tau zero of GIRK1/GIRK4 activated with guanosine 5'-O-(3-thiotriphosphate) was 1.3 +/- 0.1 ms (100%), and the channel activity (NP0) was almost fivefold lower. These changes in channel kinetics did not occur in the presence of sodium orthovanadate (3 mM), an inhibitor of phosphatase. Addition of 1 mM ATP, but not adenylimidodiphosphate, to inside-out patches resulted in increases in NP zero (4.8-fold) and the open-time duration of GIRK1/GIRK4, such that tau zero were 1.2 +/- 0.2 (32%) and 6.2 +/- 0.6 ms (68%). Single channel conductances were unchanged (34 +/- 1 pS). Cytosolic extract from atria, but not oocytes, could reverse these effects of ATP. These results provide further evidence that the antagonistic modulation of G protein-gated K+ channels by ATP and the atrial cytosolic protein produces the early rapid desensitization in atrial cells. In oocytes the ATP-dependent step is dominant and thus provides a major component of the total GIRK1/GIRK4 current activated by ACh.


Long-acting depot formulation of luprolide acetate as a method of hypothalamic down regulation for controlled ovarian hyperstimulation and oocyte production in Macaca fascicularis.
Reproductive function in some nonhuman primate species parallels that of the human. As a result, studies addressing aspects of reproductive function primarily involve the use of nonhuman primate models. The objective of the present study was to assess the efficiency of two hypothalamic down-regulation techniques combined with a single controlled ovarian hyperstimulation protocol for mature oocyte production in the cynomolgus macaque (Macaca fascicularis). Hypothalamic GnRH down regulation was first induced using the clinical long protocol of the short-acting GnRH-agonist luprolide acetate combined with controlled ovarian hyperstimulation and oocyte retrieval. Resulting oocyte yield and maturity with this regimen was insufficient for further evaluation of oocyte competency. Hypothalamic down regulation was induced in the second experiment using the long-acting depot formulation of luprolide acetate in conjunction with controlled ovarian hyperstimulation. This regimen allowed for the consistently efficient production of oocytes (15.5 oocytes per oocyte retrieval) and an oocyte maturity rate of 56%. Oocyte competence, as determined by the ability to undergo fertilization or parthenogenic activation and to reach specific cleavage stages at appropriate time intervals, was evaluated. Intracytoplasmic sperm injection resulted in a 59% fertilization rate and a 91% cleavage rate. Parthenogenic activation resulted in a 70% activation rate and an 86% cleavage rate. These data suggest that use of the long-acting form of luprolide acetate in conjunction with controlled ovarian hyperstimulation results in the production of competent, mature oocytes and allows the efficient use of nonhuman primate resources in studies of reproductive function in cynomolgus macaques.


Antimalarial efficacy of a quantified extract of Nauclea pobeguinii stem bark in human adult volunteers with diagnosed uncomplicated falciparum malaria. Part 1: a clinical phase IIA trial.
The aim of this phase IIA clinical trial was to assess the efficacy of an 80 % ethanolic quantified extract (containing 5.6 % strictosamide as the putative active constituent) from Nauclea pobeguinii stem bark denoted as PR 259 CT1 in a small group of adult patients diagnosed with uncomplicated falciparum malaria. Results obtained from a phase I clinical trial on healthy male volunteers indicated that the oral administration during meals of two 500 mg capsules three times daily (each eight hours) during seven days was well tolerated and showed only mild and self-resolving adverse effects. This PR 259 CT1 drug regimen was obtained by mathematical conversion of animal doses obtained in several in vivo studies in mice to human equivalent doses as in falciparum malaria patients. The phase IIA study was an open cohort study in eleven appraisable adult patients suffering from proven Plasmodium falciparum malaria. The study was specifically designed to assess the efficacy of PR 259 CT1 administered with a dose regimen of two 500 mg capsules three times daily for three days, followed by outpatient treatment of one 500 mg capsule three times daily for the next four days, in order to prove that this therapeutic dose, which was calculated from animal doses, was effective to treat adult malaria patients and consequently useful for a future Phase IIB clinical trial. This study would then substitute a dose-escalating trial, which in general is used to find the appropriate dose for clinical studies. The phase IIA clinical trial was carried out according to the WHO 2003 14-day test, and the results revealed that all eleven patients were completely cleared of parasitemia and fever on days 3, 7, and 14 except for one patient, who experienced a recurrence of parasitemia at days 7 until 14. Besides this adequate clinical and parasitological response (ACPR), this trial also demonstrated that PR 259 CT1 was well tolerated with only mild and self-resolving adverse effects including fatigue and headache, which were in accordance with those found in the phase I clinical trial. Moreover, all symptoms progressively disappeared, and no symptoms were observed on day 14. Although the number of patients included in this study was rather limited, the statistical analysis nevertheless suggested the efficacy and tolerability of PR 259 CT1, which indicated that this herbal medicinal product might be considered as a putative candidate for a large scale clinical trial.


Reliability and Generalizability of Similarity-Based Fusion of MEG and fMRI Data in Human Ventral and Dorsal Visual Streams.
To build a representation of what we see, the human brain recruits regions throughout the visual cortex in cascading sequence. Recently, an approach was proposed to evaluate the dynamics of visual perception in high spatiotemporal resolution at the scale of the whole brain. This method combined functional magnetic resonance imaging (fMRI) data with magnetoencephalography (MEG) data using representational similarity analysis and revealed a hierarchical progression from primary visual cortex through the dorsal and ventral streams. To assess the replicability of this method, we here present the results of a visual recognition neuro-imaging fusion experiment and compare them within and across experimental settings. We evaluated the reliability of this method by assessing the consistency of the results under similar test conditions, showing high agreement within participants. We then generalized these results to a separate group of individuals and visual input by comparing them to the fMRI-MEG fusion data of Cichy et al (2016), revealing a highly similar temporal progression recruiting both the dorsal and ventral streams. Together these results are a testament to the reproducibility of the fMRI-MEG fusion approach and allows for the interpretation of these spatiotemporal dynamic in a broader context.


Activity of Boscalid, Fenhexamid, Fluazinam, Fludioxonil, and Vinclozolin on Growth of Sclerotinia minor and S. sclerotiorum and Development of Lettuce Drop.
Sclerotinia drop is a major disease of lettuce caused by two soilborne fungi, Sclerotinia minor and S. sclerotiorum. Fungicides such as dicloran (Botran), iprodione (Rovral), and vinclozolin (Ronilan) are currently available in the United States to manage this disease. Studies were conducted to investigate the relative effect of some new fungicides, including boscalid, fenhexamid, fluazinam, and fludioxonil, in comparison with vinclozolin, on growth of S. minor and S. sclerotiorum in agar plate tests as well as control of lettuce drop in the field. At a rate of 0.001 μg/ml, all tested compounds only suppressed mycelial growth of either pathogen from 0 to 20%. At 0.01 μg/ml, mycelial growth of S. minor was reduced 82 to 84% by fludioxonil and fluazinam and only 1 to 16% by boscalid, fenhexamid, and vinclozolin. At the same rate, mycelial growth of S. sclerotiorum was reduced 78% by fluazinam and from 0 to 12% by boscalid, fludioxonil, fenhexamid, and vinclozolin. At 0.1 μg/ml, all tested chemistries except vinclozolin inhibited mycelial growth of S. minor from 70 to 98%, whereas growth of S. sclerotiorum was suppressed 95 to 99% by fludioxonil and fluazinam, significantly less (40 to 47%) by boscalid and fenhexamid, and not at all by vinclozolin. At a rate of 1.0 μg/ml, all tested fungicides reduced mycelial growth of S. minor and S. sclerotiorum from 87 to 100% and 77 to 100%, respectively. Mycelial growth emerging from sclerotia of S. minor was reduced from 98 to 100% by all fungicides tested at a rate of 1.0 μg/ml, whereas growth from sclerotia of S. sclerotiorum was suppressed from 90 to 96% by fenhexamid, fludioxonil, fluazinam, and vinclozolin. In lettuce plots infested with S. minor, boscalid and fluazinam provided the highest level of disease control, significantly greater than that achieved with fenhexamid, fludioxonil, and vinclozolin. In the presence of S. sclerotiorum, the highest degree of disease suppression occurred with application of fluazinam, fludioxonil, and vinclozolin, whereas the least effective compound was fenhexamid. Boscalid and fluazinam were more effective against lettuce drop caused by S. minor than disease caused by S. sclerotiorum.


Cooperation in USDA studies and evaluations, and full use of federal funds in nutrition assistance programs nondiscretionary provisions of the Healthy, Hunger-Free Kids Act of 2010, Public Law 111-296. Final Rule.
This final rule incorporates into the regulations governing the Programs authorized under the Richard B. Russell National School Lunch Act (NSLA) and the Child Nutrition Act of 1966 (CNA) two nondiscretionary provisions of the Healthy, Hunger-Free Kids Act of 2010 (HHFK Act). The HHFK Act requires State and local cooperation in Department of Agriculture studies and evaluations related to Programs authorized under the NSLA and the CNA. The HHFK Act also amends the NSLA to stipulate that Federal funds must not be subject to State budget restrictions or limitations, including hiring freezes, work furloughs, and travel restrictions. This final rule amends regulations for the National School Lunch Program; the Special Milk Program for Children; the School Breakfast Program; the Summer Food Service Program; the Child and Adult Care Food Program; State Administrative Expense Funds ; the Special Supplemental Nutrition Program for Women, Infants and Children; and the WIC Farmers' Market Nutrition Program. These provisions will strengthen program integrity by ensuring that sufficient data is made available for studies and evaluations. Additionally, exempting Federal funds from State budgetary restrictions or limitations is intended to increase the ability of State agencies to administer USDA's nutrition assistance programs effectively.


N-palmitoylethanolamide Prevents Parkinsonian Phenotypes in Aged Mice.
Parkinson's disease (PD) is a neurodegenerative disease characterized by degeneration of dopaminergic neurons. Aging is a major risk factor for idiopathic PD. Several prior studies examined the neuroprotective effects of palmitoylethanolamide (PEA), alone or combined with antioxidants, in a model of PD induced by the dopaminergic toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Here, we analyzed the pretreatment effect of micronized PEA (PEAm) on neuroinflammation and neuronal cell death in the MPTP model. Male CD mice (21 months of age) were pre-treated for 60 days with PEAm. After this time, they received four intraperitoneal injections of MPTP over a 24-h period and were killed 7 days later. On the 8th day, brains were processed. Pretreatment with PEAm ameliorated behavioral deficits and the reductions in expression of tyrosine hydroxylase and dopamine transporter, while blunting the upregulation of α-synuclein and β3-tubulin in the substantia nigra after MPTP induction. Moreover, PEAm reduced proinflammatory cytokine expression and showed a pro-neurogenic effect in hippocampus. These findings propose this strategy as a valid approach to prevent neurodegenerative diseases associated with old age.


Epigallocatechin gallate acts synergistically in combination with cisplatin and designed trans-palladiums in ovarian cancer cells.
In this study, synergism in activity from the sequenced combinations of three trans-palladiums (denoted as TH5, TH6 and TH7) with green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG), as well as that with cisplatin, was investigated in a number of human ovarian tumour models as a function of sequence of administration. Cellular accumulation of platinum and palladium, and the levels of platinum-DNA and palladium-DNA binding were also determined for the 0/4 h and 0/0 h sequences of administration. The results of the study show that co-administration of cisplatin with EGCG (0/0 h) produces weak synergism in both cisplatin-sensitive (A2780) and cisplatin-resistant (A2780(cisR)) cell lines whereas (0/4 h) administration produces pronounced synergism in both. In contrast, bolus administration of EGCG with TH5, TH6 and TH7 produces marked antagonism except that with TH5, in the A2780(cisR) cell line, where a mild synergism is observed. In the case of TH5, TH6 and TH7, administration of drugs with a time gap (0/4 h or 4/0 h combinations) produces sequence-dependent synergism in both A2780 and A2780(cisR) cell lines, whereas in the case of cisplatin, marked antagonism is observed with the 4/0 h sequence of administration in the A2780 cell line. Whereas the highly synergistic 0/4 h sequence of combination of cisplatin with EGCG is found to be associated with pronounced cellular accumulation of platinum and a high level of platinum-DNA binding, no such clear trend can be seen for any of the combinations of TH5, TH6 and TH7 with EGCG. The results of the present study provide support to the idea that sequenced combinations of platinum drugs and tumour-active palladium compounds with selected phytochemicals such as EGCG may provide a means of overcoming drug resistance.


Mechanism of photogenerated reactive oxygen species and correlation with the antibacterial properties of engineered metal-oxide nanoparticles.
Oxidative stress induced by reactive oxygen species (ROS) is one of the most important antibacterial mechanisms of engineered nanoparticles (NPs). To elucidate the ROS generation mechanisms, we investigated the ROS production kinetics of seven selected metal-oxide NPs and their bulk counterparts under UV irradiation (365 nm). The results show that different metal oxides had distinct photogenerated ROS kinetics. Particularly, TiO(2) nanoparticles and ZnO nanoparticles generated three types of ROS (superoxide radical, hydroxyl radical, and singlet oxygen), whereas other metal oxides generated only one or two types or did not generate any type of ROS. Moreover, NPs yielded more ROS than their bulk counterparts likely due to larger surface areas of NPs providing more absorption sites for UV irradiation. The ROS generation mechanism was elucidated by comparing the electronic structures (i.e., band edge energy levels) of the metal oxides with the redox potentials of various ROS generation, which correctly interpreted the ROS generation of most metal oxides. To develop a quantitative relationship between oxidative stress and antibacterial activity of NPs, we examined the viability of E. coli cells in aqueous suspensions of NPs under UV irradiation, and a linear correlation was found between the average concentration of total ROS and the bacterial survival rates (R(2) = 0.84). Although some NPs (i.e., ZnO and CuO nanoparticles) released toxic ions that partially contributed to their antibacterial activity, this correlation quantitatively linked ROS production capability of NPs to their antibacterial activity as well as shed light on the applications of metal-oxide NPs as potential antibacterial agents.


A polymorphism at the translation start site of the vitamin D receptor gene is associated with the response to anti-osteoporotic therapy in postmenopausal women from southern Italy.
The present study investigated the effect of two single nucleotide polymorphisms (SNPs) of the vitamin D receptor (VDR) gene, rs1544410 A/G and rs2228570 C/T, in modulating bone mineral density (BMD) and the response to treatment with bisphosphonates or strontium ranelate in postmenopausal osteoporosis (PMO). Four hundred eighteen postmenopausal women from Southern Italy treated with bisphosphonates or strontium ranelate for three years were enrolled and stratified according to their genotype. Changes in BMD were expressed as the delta t-score (Δt-score). Allelic frequencies for rs1544410 A/GSNP were 11.2% AA, 50.0% GA and 38.8% GG; for rs2228570 C/TSNP were 54.8% CC, 39.5% TC and 5.7% TT. TT carriers showed a lower t-score than TC and CC (both p < 0.02) genotypes and were more responsive to the therapy when compared to both TC (p < 0.02) and CC (p < 0.05) carriers. Specifically, TT carriers receiving alendronate demonstrated a significant improvement of the Δt-score compared to TC and CC (both p < 0.0001) carriers. After adjustment for confounders, the Δt-score showed evidence of a statistically significant positive association with TT in all treatments considered. Therapy response was independent of rs1544410 A/G SNP; instead, rs2228570 C/TSNP was associated with a better response to antiresorptive treatment, thus suggesting that the therapy for PMO should be personalized.


Comparison of liver biopsy and noninvasive techniques for liver fibrosis assessment in patients infected with HCV-genotype 4 in Egypt.
In Egypt, as elsewhere, liver biopsy (LB) remains the gold standard to assess liver fibrosis in chronic hepatitis C (CHC) and is required to decide whether a treatment should be proposed. Many of its disadvantages have led to develop noninvasive methods to replace LB. These new methods should be evaluated in Egypt, where circulating virus genotype 4 (G4), increased body mass index and co-infection with schistosomiasis may interfere with liver fibrosis assessment. Egyptian CHC-infected patients with G4 underwent a LB, an elastometry measurement (Fibroscan(©)), and serum markers (APRI, Fib4 and Fibrotest(©)). Patients had to have a LB ≥15 mm length or ≥10 portal tracts with two pathologists blinded readings to be included in the analysis. Patients with hepatitis B virus co-infection were excluded. Three hundred and twelve patients are reported. The performance of each technique for distinguishing F0F1 vs F2F3F4 was compared. The area under receiver operating characteristic curves was 0.70, 0.76, 0.71 and 0.75 for APRI, Fib-4, Fibrotest© and Fibroscan©, respectively (no influence of schistosomiasis was noticed). An algorithm using the Fib4 for identifying patients with F2 stage or more reduced by nearly 90% the number of liver biopsies. Our results demonstrated that noninvasive techniques were feasible in Egypt, for CHC G4-infected patients. Because of its validity and its easiness to perform, we believe that Fib4 may be used to assess the F2 threshold, which decides whether treatment should be proposed or delayed.


An arthroconidial-spherule antigen of Coccidioides immitis: differential expression during in vitro fungal development and evidence for humoral response in humans after infection or vaccination.
A 33-kDa protein antigen purified from spherules of Coccidioides immitis was analyzed for ultrastructural localization and for binding to serum antibodies from infected or immunized humans. By using colloidal gold detection of affinity-purified anti-33-kDa protein antibodies, electron photomicrographs showed binding to the inner cell wall of arthroconidia and spherules and to the septa and glycocalyx surrounding endospores. Enzyme immunoassay measurements also demonstrated that the antigen was most abundant in mature spherules. Of 37 patients with coccidioidomycosis but without concurrent human immunodeficiency virus infections, all but 2 demonstrated immunoglobulin M (IgM) (usually with early infection) or IgG antibodies for the 33-kDa antigen. In contrast, only one of four HIV-infected patients with active coccidioidal infections demonstrated antibody. On the other hand, 107 of 108 patients without evident coccidioidomycosis and 15 of 16 patients with histoplasmosis did not have similar antibodies, indicating a high degree of specificity. Immunization of humans with a spherule vaccine produced IgM responses to this antigen that were not evident in placebo recipients.


Use of a novel antigen expressing system to study the Salmonella enterica serovar Typhi protein recognition by T cells.
Salmonella enterica serovar Typhi (S. Typhi), the causative agent of the typhoid fever, is a pathogen of great public health importance. Typhoid vaccines have the potential to be cost-effective measures towards combating this disease, yet the antigens triggering host protective immune responses are largely unknown. Given the key role of cellular-mediated immunity in S. Typhi protection, it is crucial to identify S. Typhi proteins involved in T-cell responses. Here, cells from individuals immunized with Ty21a typhoid vaccine were collected before and after immunization and used as effectors. We also used an innovative antigen expressing system based on the infection of B-cells with recombinant Escherichia coli (E. coli) expressing one of four S. Typhi gene products (i.e., SifA, OmpC, FliC, GroEL) as targets. Using flow cytometry, we found that the pattern of response to specific S. Typhi proteins was variable. Some individuals responded to all four proteins while others responded to only one or two proteins. We next evaluated whether T-cells responding to recombinant E. coli also possess the ability to respond to purified proteins. We observed that CD4+ cell responses, but not CD8+ cell responses, to recombinant E. coli were significantly associated with the responses to purified proteins. Thus, our results demonstrate the feasibility of using an E. coli expressing system to uncover the antigen specificity of T-cells and highlight its applicability to vaccine studies. These results also emphasize the importance of selecting the stimuli appropriately when evaluating CD4+ and CD8+ cell responses.


Menstrual disorders in patients with chronic renal failure.
Forty females, age 14 to 35 years (mean 28.6 years) with chronic renal failure (CRF) were included in the study. Their menstrual patterns were noted. The function of their hypothalamo-pituitary-ovarian axis was assessed by the serum levels of follicle stimulating hormone (FSH), Luteinising hormone (LH), prolactin (PrL), estradiol (E2) and progesterone (P) at different phases of the menstrual cycle in patients who continued to have normal menses (Group 1) and at weekly intervals for six weeks in patients with menstrual disturbances (Group II). The mean hormone levels during the initial contact Luteal phase in group I were FSH 12.0 IU/L (N, 1.0-3.0 IU/L), LH 1.8IU/L (N 1.5-101U/L), PrL 652mIU/L (N, 100-600 mIU/L) mE2 160 pmol/L (N 400-1400 pmol/L) and P5 nmol/L (N 14-60 nmol/L) for group I. Corresponding values for group II were 1.2, 10.3, 250, 600 and 3.0 in relevant units. All patients (fourteen) with end stage renal disease (ESRD) had amenorrhoae. On the other hand, most patients with stable CRF (22/26) had normal menses. Following initiation of therapy (conservative or dialytic), there was no significant alteration in the hormonal profile or menstrual pattern. We conclude that other factors apart from the hormonal imbalances, may be responsible for the menstrual disturbances noted in patients with CRF.


Regulation of impulsive and aggressive behaviours by a novel lncRNA.
High impulsive and aggressive traits associate with poor behavioural self-control. Despite their importance in predicting behavioural negative outcomes including suicide, the molecular mechanisms underlying the expression of impulsive and aggressive traits remain poorly understood. Here, we identified and characterized a novel long noncoding RNA (lncRNA), acting as a regulator of the monoamine oxidase A (MAOA) gene in the brain, and named it MAOA-associated lncRNA (MAALIN). Our results show that in the brain of suicide completers, MAALIN is regulated by a combination of epigenetic mechanisms including DNA methylation and chromatin modifications. Elevated MAALIN in the dentate gyrus of impulsive-aggressive suicides was associated with lower MAOA expression. Viral overexpression of MAALIN in neuroprogenitor cells decreased MAOA expression while CRISPR-mediated knock out resulted in elevated MAOA expression. Using viral-mediated gene transfer, we confirmed that MAALIN in the hippocampus significantly decreases MAOA expression and exacerbates the expression of impulsive-aggressive behavioural traits in CD1 aggressive mice. Overall, our findings suggest that variations in DNA methylation mediate the differential expression of a novel lncRNA that acts on MAOA expression to regulate impulsive-aggressive behaviours.


Prevalence and Risk-Markers of Self-Harm in Autistic Children and Adults.
Self-harm is purportedly common in autistic individuals, but under-researched, particularly in younger samples and those without intellectual disability. This study aimed to describe prevalence, profile and correlates of self-harm in autistic individuals without impairments in adaptive functioning. Parents of autistic participants (n = 83) completed questionnaires regarding the presence/topography of self-harm, demographic characteristics, autism severity, age of diagnosis, affect, activity levels and repetitive behaviour. 24.10% of participants engaged in self-harm. Self-harm was associated with significantly higher levels of impulsivity, over-activity, negative affect, compulsive behaviour and insistence on sameness. Low mood and overactivity/impulsivity predicted the presence of self-harm, with the model correctly classifying 82.9% of cases. Findings highlight a role for impaired behavioural inhibition and low mood in the aetiological mechanisms underpinning self-harm in autism.


Factors impacting employment for people with autism spectrum disorder: A scoping review.
The aim of this study is to holistically synthesise the extent and range of literature relating to the employment of individuals with autism spectrum disorder. Database searches of Medline, CINAHL, PsychINFO, Scopus, ERIC, Web of Science and EMBASE were conducted. Studies describing adults with autism spectrum disorder employed in competitive, supported or sheltered employment were included. Content analysis was used to identify the strengths and abilities in the workplace of employees with autism spectrum disorder. Finally, meaningful concepts relating to employment interventions were extracted and linked to the International Classification of Functioning, Disability and Health Core Sets for autism spectrum disorder. The search identified 134 studies for inclusion with methodological quality ranging from limited to strong. Of these studies, only 36 evaluated employment interventions that were coded and linked to the International Classification of Functioning, Disability and Health, primarily focusing on modifying autism spectrum disorder characteristics for improved job performance, with little consideration of the impact of contextual factors on work participation. The International Classification of Functioning, Disability and Health Core Sets for autism spectrum disorder are a useful tool in holistically examining the employment literature for individuals with autism spectrum disorder. This review highlighted the key role that environmental factors play as barriers and facilitators in the employment of people with autism spectrum disorder and the critical need for interventions which target contextual factors if employment outcomes are to be improved.


Isolated truncus arteriosus associated with a mutation in the plexin-D1 gene.
Truncus arteriosus accounts for approximately 1% of congenital heart defects and the cause of isolated non-syndromic truncus arteriosus is largely unknown. In order to identify the underlying molecular defect in a consanguineous family with recurrent tuncus arteriosus, homozygosity mapping followed by whole exome sequencing was performed. This resulted in the identification of a homozygous mutation, Arg1299Cys, in the PLXND1 gene. The mutation affected a highly conserved residue, segregated with the disease in the family and was absent from available SNP databases and ethnic matched controls. in silico comparative modeling revealed that the mutation resides in the N-terminal segment of the human plexin-D1 intracellular region which interacts with the catalytic GTPase-activating protein homology region. The mutation likely destabilizes the intracellular region, perturbing its anchoring and catalytic activity. The phenotype in human PLXND1 mutation is closely related to that of knockout mice for PLXND1, its co-receptor neuropilin-1 or its ligand SEMA3C. It is therefore suggested that SEMA3C signaling, propagated through the heterodimer receptor plexin-D1/neuropilin, is important for truncus arteriosus septation. Confirmation of this observation will require the identification of PLXND1 mutations in additional patients. Exome analysis is valuable for molecular investigation of single patients with congenital heart defects in whom chromosomal copy number variants have been excluded.


Fear Spreading Across Senses: Visual Emotional Events Alter Cortical Responses to Touch, Audition, and Vision.
Attention and perception are potentiated for emotionally significant stimuli, promoting efficient reactivity and survival. But does such enhancement extend to stimuli simultaneously presented across different sensory modalities? We used functional magnetic resonance imaging in humans to examine the effects of visual emotional signals on concomitant sensory inputs in auditory, somatosensory, and visual modalities. First, we identified sensory areas responsive to task-irrelevant tones, touches, or flickers, presented bilaterally while participants attended to either a neutral or a fearful face. Then, we measured whether these responses were modulated by the emotional content of the face. Sensory responses in primary cortices were enhanced for auditory and tactile stimuli when these appeared with fearful faces, compared with neutral, but striate cortex responses to the visual stimuli were reduced in the left hemisphere, plausibly as a consequence of sensory competition. Finally, conjunction and functional connectivity analyses identified 2 distinct networks presumably responsible for these emotional modulatory processes, involving cingulate, insular, and orbitofrontal cortices for the increased sensory responses, and ventrolateral prefrontal cortex for the decreased sensory responses. These results suggest that emotion tunes the excitability of sensory systems across multiple modalities simultaneously, allowing the individual to adaptively process incoming inputs in a potentially threatening environment.


Obesity Accelerates Alzheimer-Related Pathology in APOE4 but not APOE3 Mice.
Alzheimer's disease (AD) risk is modified by both genetic and environmental risk factors, which are believed to interact to cooperatively modify pathogenesis. Although numerous genetic and environmental risk factors for AD have been identified, relatively little is known about potential gene-environment interactions in regulating disease risk. The strongest genetic risk factor for late-onset AD is the ε4 allele of apolipoprotein E (APOE4). An important modifiable risk factor for AD is obesity, which has been shown to increase AD risk in humans and accelerate development of AD-related pathology in rodent models. Potential interactions between APOE4 and obesity are suggested by the literature but have not been thoroughly investigated. In the current study, we evaluated this relationship by studying the effects of diet-induced obesity (DIO) in the EFAD mouse model, which combines familial AD transgenes with human APOE3 or APOE4. Male E3FAD and E4FAD mice were maintained for 12 weeks on either a control diet or a Western diet high in saturated fat and sugars. We observed that metabolic outcomes of DIO were similar in E3FAD and E4FAD mice. Importantly, our data showed a significant interaction between diet and APOE genotype on AD-related outcomes in which Western diet was associated with robust increases in amyloid deposits, β-amyloid burden, and glial activation in E4FAD but not in E3FAD mice. These findings demonstrate an important gene-environment interaction in an AD mouse model that suggests that AD risk associated with obesity is strongly influenced by APOE genotype.