Frequent foamy virus infection in free-living chimpanzees of the Taï National Park (Côte d'Ivoire).
Foamy viruses are frequently found in non-human primates and apes in captivity. However, data on simian foamy virus (SFV) infection in apes from the wild are limited. Necropsy specimens were collected from 14 West African chimpanzees (Pan troglodytes verus) from three communities in the Taï National Park, Côte d'Ivoire. PCR analysis revealed SFV-related int- and env-specific sequences in 12/14 chimpanzees. Two young chimpanzees were not infected. Plasma from 'PCR-positive' chimpanzees reacted against Pr71/74(gag) in Western blot analysis. Phylogenetic analysis demonstrated clustering of all analysed sequences with SFVcpz previously identified from the other P. troglodytes verus, although interestingly the sequences were diverse and no grouping according to a particular animal community was observed. The body compartments of two infected animals were examined and found to contain SFV sequences. Frequent SFV infections in chimpanzees from this area significantly increase the potential risk of zoonotic transmission to rural populations through direct contact, hunting and consumption of bush meat.


Charge separation and isolation in strong water droplet impacts.
Charge separation in condensed matter after strong impacts is a general and intriguing phenomenon in nature, which is often identified and described but not necessarily well understood in terms of a quantitative mechanistic picture. Here we show that charge separation naturally occurs if water droplets/clusters or ice particles with embedded charge carriers, e.g., ions, encounter a high energy impact with subsequent dispersion - even if the involved kinetic energy is significantly below the molecular ionization energy. We find that for low charge carrier concentrations (c < 0.01 mol L(-1)) a simple statistical Poisson model describes the charge distribution in the resulting molecular "fragments" or aggregates. At higher concentrations Coulomb interactions between the charge carriers become relevant, which we describe by a Monte Carlo approach. Our models are compared to experimental data for strong (laser) impacts on liquid micro beams and discussed for the charge generation in cluster-impact mass spectrometry on cosmic dust detectors where particle kinetic energies are below the plasma threshold. Taken together, a simple and intuitive but quantitative microscopic model is obtained, which may contribute to the understanding of a larger range of phenomena related to charge generation and separation in nature.


Gaps in using bronchodilators, inhaled corticosteroids and influenza vaccine among 23 high- and low-income sites.
Increasing access to essential respiratory medicines and influenza vaccination has been a priority for over three decades. Their use remains low in low- and middle-income countries (LMICs), where little is known about factors influencing use, or about the use of influenza vaccination for preventing respiratory exacerbations. We estimated rates of regular use of bronchodilators, inhaled corticosteroids and influenza vaccine, and predictors for use among 19 000 adults in 23 high-income countries (HICs) and LMIC sites. Bronchodilators, inhaled corticosteroids and influenza vaccine were used significantly more in HICs than in LMICs, after adjusting for similar clinical needs. Although they are used more commonly by people with symptomatic or severe respiratory disease, the gap between HICs and LMICs is not explained by the prevalence of chronic obstructive pulmonary disease or doctor-diagnosed asthma. Site-specific factors are likely to influence use differently. The gross national income per capita for the country is a strong predictor for use of these treatments, suggesting that economics influence under-treatment. We still need a better understanding of determinants for the low use of essential respiratory medicines and influenza vaccine in low-income settings. Identifying and addressing these more systematically could improve the access and use of effective treatments.


Rapid polymerase chain reaction-restriction fragment length polymorphism method for discrimination of the two Atlantic cryptic deep-sea species of scabbardfish.
The present investigation provides an efficient diagnostic method based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis to discriminate between two cryptic species of scabbardfish, Aphanopus carbo and A. intermedius, with commercial relevance in several European fish markets. Two DNA fragments from the mtDNA, including control region and partial cytochrome oxidase subunit I genes of about 1100 bp and 700 bp, respectively, were isolated by PCR amplification. Digestion of the amplicon including the control region with HaeII and the amplicon including the COI gene with Sau3AI restriction enzymes allowed an unequivocal discrimination between the two scabbardfish species. This PCR-RFLP method allowed a clear and rapid discrimination of the trichiurid species studied.


The Circuit Theory Behind Coupled-Mode Magnetic Resonance-Based Wireless Power Transmission.
Inductive coupling is a viable scheme to wirelessly energize devices with a wide range of power requirements from nanowatts in radio frequency identification tags to milliwatts in implantable microelectronic devices, watts in mobile electronics, and kilowatts in electric cars. Several analytical methods for estimating the power transfer efficiency (PTE) across inductive power transmission links have been devised based on circuit and electromagnetic theories by electrical engineers and physicists, respectively. However, a direct side-by-side comparison between these two approaches is lacking. Here, we have analyzed the PTE of a pair of capacitively loaded inductors via reflected load theory (RLT) and compared it with a method known as coupled-mode theory (CMT). We have also derived PTE equations for multiple capacitively loaded inductors based on both RLT and CMT. We have proven that both methods basically result in the same set of equations in steady state and either method can be applied for short- or midrange coupling conditions. We have verified the accuracy of both methods through measurements, and also analyzed the transient response of a pair of capacitively loaded inductors. Our analysis shows that the CMT is only applicable to coils with high quality factor (Q) and large coupling distance. It simplifies the analysis by reducing the order of the differential equations by half compared to the circuit theory.


Ensembles of engineered cardiac tissues for physiological and pharmacological study: heart on a chip.
Traditionally, muscle physiology experiments require multiple tissue samples to obtain morphometric, electrophysiological, and contractility data. Furthermore, these experiments are commonly completed one at a time on cover slips of single cells, isotropic monolayers, or in isolated muscle strips. In all of these cases, variability of the samples hinders quantitative comparisons among experimental groups. Here, we report the design of a "heart on a chip" that exploits muscular thin film technology--biohybrid constructs of an engineered, anisotropic ventricular myocardium on an elastomeric thin film--to measure contractility, combined with a quantification of action potential propagation, and cytoskeletal architecture in multiple tissues in the same experiment. We report techniques for real-time data collection and analysis during pharmacological intervention. The chip is an efficient means of measuring structure-function relationships in constructs that replicate the hierarchical tissue architectures of laminar cardiac muscle.


Stress proteins on the yeast cell surface determine resistance to osmotin, a plant antifungal protein.
Strains of the yeast Saccharomyces cerevisiae differ in their sensitivities to tobacco osmotin, an antifungal protein of the PR-5 family. However, cells sensitive to tobacco osmotin showed resistance to osmotin-like proteins purified from the plant Atriplex nummularia, indicating a strict specificity between the antifungal protein and its target cell. A member of a gene family encoding stress proteins induced by heat and nitrogen limitation, collectively called Pir proteins, was isolated among the genes that conveyed resistance to tobacco osmotin to a susceptible strain. We show that overexpression of Pir proteins increased resistance to osmotin, whereas simultaneous deletion of all PIR genes in a tolerant strain resulted in sensitivity. Pir proteins have been immunolocalized to the cell wall. The enzymatic digestion of the cell wall of sensitive and resistant cells rendered spheroplasts equally susceptible to the cytotoxic action of tobacco osmotin but not to other osmotin-like proteins, indicating that the cell membrane interacts specifically with osmotin and facilitates its action. Our results demonstrate that fungal cell wall proteins are determinants of resistance to antifungal PR-5 proteins.


Toxicity and therapeutic efficacy of high-dose interleukin 2. In vivo infusion of antibody to NK-1.1 attenuates toxicity without compromising efficacy against murine leukemia.
In the current study we used the therapy of established murine leukemia to identify the lymphocyte subsets responsible for toxicity and for therapeutic efficacy of high-dose IL-2. Initial results confirmed that high-dose IL-2 induces marked proliferation of a variety of host cells, including NK cells, Lyt-2+ T cells, L3T4+ T cells, and B cells. Infusion of antibody to NK-1.1 depleted NK-1.1+ cells in vivo and greatly reduced the toxicity of IL-2, but did not decrease therapeutic efficacy. By marked contrast, depletion of host T cells, either Lyt-2+ or L3T4+, had no effect on toxicity but greatly reduced therapeutic efficacy. The requirement for host T cells for the curative effect of IL-2 gives credence to the possibility that substantial efficacy of high-dose IL-2 against established malignancy may require existent host antitumor immunity. Since the human tumors that have been shown to have the most substantial responses to IL-2 (i.e., malignant melanoma and renal cell carcinoma) are those long considered to be immunogenic in the autochthonous host, the current study predicts that for these, as well as other immunogenic human tumors, it should be possible to decrease the toxicity and thus increase the therapeutic index of IL-2 by selectively depleting NK cells in vivo.


Dehydroepiandrosterone (DHEA) prevents and reverses chronic hypoxic pulmonary hypertension.
Pulmonary artery (PA) hypertension was studied in a chronic hypoxic-pulmonary hypertension model (7-21 days) in the rat. Increase in PA pressure (measured by catheterism), cardiac right ventricle hypertrophy (determined by echocardiography), and PA remodeling (evaluated by histology) were almost entirely prevented after oral dehydroepiandrosterone (DHEA) administration (30 mg/kg every alternate day). Furthermore, in hypertensive rats, oral administration, or intravascular injection (into the jugular vein) of DHEA rapidly decreased PA hypertension. In PA smooth muscle cells, DHEA reduced the level of intracellular calcium (measured by microspectrofluorimetry). The effect of DHEA appears to involve a large conductance Ca2+-activated potassium channel (BKCa)-dependent stimulatory mechanism, at both function and expression levels (isometric contraction and Western blot), via a redox-dependent pathway. Voltage-gated potassium (Kv) channels also may be involved because the antagonist 4-amino-pyridine blocked part of the DHEA effect. The possible pathophysiological and therapeutic significance of the results is discussed.


Seroepidemiological Prevalence of Multiple Species of Filoviruses in Fruit Bats (Eidolon helvum) Migrating in Africa.
Fruit bats are suspected to be a natural reservoir of filoviruses, including Ebola and Marburg viruses. Using an enzyme-linked immunosorbent assay based on the viral glycoprotein antigens, we detected filovirus-specific immunoglobulin G antibodies in 71 of 748 serum samples collected from migratory fruit bats (Eidolon helvum) in Zambia during 2006-2013. Although antibodies to African filoviruses (eg, Zaire ebolavirus) were most prevalent, some serum samples showed distinct specificity for Reston ebolavirus, which that has thus far been found only in Asia. Interestingly, the transition of filovirus species causing outbreaks in Central and West Africa during 2005-2014 seemed to be synchronized with the change of the serologically dominant virus species in these bats. These data suggest the introduction of multiple species of filoviruses in the migratory bat population and point to the need for continued surveillance of filovirus infection of wild animals in sub-Saharan Africa, including hitherto nonendemic countries.


Neither property right nor heroic gift, neither sacrifice nor aporia: the benefit of the theoretical lens of sharing in donation ethics.
Two ethical frameworks have dominated the discussion of organ donation for long: that of property rights and that of gift-giving. However, recent years have seen a drastic rise in the number of philosophical analyses of the meaning of giving and generosity, which has been mirrored in ethical debates on organ donation and in critical sociological, anthropological and ethnological work on the gift metaphor in this context. In order to capture the flourishing of this field, this article distinguishes between four frameworks for thinking about bodily exchanges in medicine: those of property rights, heroic gift-giving, sacrifice, and gift-giving as aporia. These frameworks represent four different ways of making sense of donation of organs as well as tissue, gametes and blood, draw on different conceptions of the relations between the self and the other, and bring out different ethical issues as core ones. The article presents these frameworks, argues that all of them run into difficulties when trying to make sense of reciprocity and relational interdependence in donation, and shows how the three gift-giving frameworks (of heroism, sacrifice and aporia) hang together in a critical discussion about what is at stake in organ donation. It also presents and argues in favour of an alternative intercorporeal framework of giving-through-sharing that more thoroughly explicates the gift metaphor in the context of donation, and offers tools for making sense of relational dimensions of live and post mortem donations.


Automated anatomical interpretation of ion distributions in tissue: linking imaging mass spectrometry to curated atlases.
Imaging mass spectrometry (IMS) has become a prime tool for studying the distribution of biomolecules in tissue. Although IMS data sets can become very large, computational methods have made it practically feasible to search these experiments for relevant findings. However, these methods lack access to an important source of information that many human interpretations rely upon: anatomical insight. In this work, we address this need by (1) integrating a curated anatomical data source with an empirically acquired IMS data source, establishing an algorithm-accessible link between them and (2) demonstrating the potential of such an IMS-anatomical atlas link by applying it toward automated anatomical interpretation of ion distributions in tissue. The concept is demonstrated in mouse brain tissue, using the Allen Mouse Brain Atlas as the curated anatomical data source that is linked to MALDI-based IMS experiments. We first develop a method to spatially map the anatomical atlas to the IMS data sets using nonrigid registration techniques. Once a mapping is established, a second computational method, called correlation-based querying, gives an elementary demonstration of the link by delivering basic insight into relationships between ion images and anatomical structures. Finally, a third algorithm moves further beyond both registration and correlation by providing automated anatomical interpretation of ion images. This task is approached as an optimization problem that deconstructs ion distributions as combinations of known anatomical structures. We demonstrate that establishing a link between an IMS experiment and an anatomical atlas enables automated anatomical annotation, which can serve as an important accelerator both for human and machine-guided exploration of IMS experiments.


Nutrition in the United States, 1900 to 1974.
Food and nutrient intakes are being examined for possible relationship to the occurrence or the progress of various types of cancer in man. This paper describes the state of our knowledge on the nutritional status of various population groups in the United States and on the food consumption patterns and their nutritional consequences during the 1900's. Only a few medical studies of nutritional status have been conducted on a national or even a regional basis. These extend back less than 2 decades. Dietary information is available back to the 1930's and statistical data on use of food in the United States can be found beginning with 1909. Sources of information are suggested for the use of and detailed study by the epidemiologist.


Urbanization and human health.
Urbanization involves a physical change in which increasing proportions of populations live in urban settings, however defined. It also implies considerable changes in the ways in which these people live, how they earn their livelihoods, the food which they eat, and the wide range of environmental factors to which they are exposed. There is another underlying assumption that, increasingly, urban populations will be more healthy than their rural counterparts and that higher levels of urbanization will equate with better health status. This paper discusses some of the assumptions underlying this contention. It takes issue with certain of them, particularly the assumption that urbanization affects the health of all residents equally. It is manifestly evident that in many cities, particularly in the developing world, the poor are exposed to greater risks and have much lower health status than their richer neighbours. In addition, whilst urban residents may theoretically have a better access to health care and services than do residents in many rural areas, and whilst many indicators of health do appear better in more highly urbanized societies than ones less so, there are caveats. The paper introduces the concept of epidemiological transition, which suggests that, whilst life expectancy might be higher in many urbanized countries and in certain cities, the inhabitants are often merely suffering from different forms of ill-health, often chronic or degenerative, rather than infective. In certain cities in middle-income countries, residents, particularly the poor, are exposed to a double risk of both infection and chronic degenerative ailments. The paper concludes with a consideration of more general recent statements from the World Health Organization among others, on the impact of urbanization on health. The 'Healthy Cities' project is also discussed. WHO identifies a range of general determinants of urban health: physical, social, cultural and environmental. Many represent the by-products of modernization and especially industrialization. It is emphasized that urbanization, and the concentration of human beings into new areas in particular, can bring exposure to new risk factors for large numbers of people. The growth of infectious and parasitic disease in some urban settings must therefore be recognized, as must the emergence of chronic diseases, with the concomitant need for investment in new types of health and social care. However, a number of constraints militate against the achievement of improved urban health, especially in developing countries.(ABSTRACT TRUNCATED AT 400 WORDS)


Comparison of necrotizing fasciitis and sepsis caused by Vibrio vulnificus and Staphylococcus aureus.
Vibrio vulnificus can cause a rapidly progressive fatal soft-tissue infection. Staphylococcus aureus is the most common cause of skin and soft-tissue infections reported worldwide, and, in particular, methicillin-resistant Staphylococcus aureus has emerged as the most common isolate in emergency departments. The purposes of the present study were to compare the specific characteristics of Vibrio vulnificus and Staphylococcus aureus infections and to compare the clinical outcomes of Vibrio vulnificus, methicillin-resistant Staphylococcus aureus, and methicillin-sensitive Staphylococcus aureus necrotizing infections. One hundred and fifteen patients with necrotizing fasciitis caused by Vibrio vulnificus (sixty patients) or Staphylococcus aureus (fifty-five patients) were retrospectively reviewed over a six-year period. Differences in mortality, patient characteristics, clinical presentations, laboratory data, and hospital course were compared between the Vibrio vulnificus and Staphylococcus aureus groups. Nineteen patients (including eleven in the Vibrio vulnificus group and eight in the Staphylococcus aureus group) died, resulting in a mortality rate of 16.5%. We found significant differences between the two groups with regard to hypotension, fever, the interval between contact and admission, the interval between the diagnosis of necrotizing fasciitis and the first operation, and admission to the intensive care unit. The patients in the Vibrio vulnificus group had significantly lower total white blood-cell counts, higher banded white blood-cell counts, and lower platelet counts as compared with those in the Staphylococcus aureus group. The proportion of patients who were hypotensive (as indicated by a systolic blood pressure of ≤ 90 mm Hg) was significantly greater in the methicillin-resistant Staphylococcus aureus subgroup than in the methicillin-sensitive Staphylococcus aureus subgroup. Patients with hepatic dysfunction were significantly more likely to have Vibrio vulnificus infection, and those with diabetes mellitus were significantly more likely to have Staphylococcus aureus infection. Necrotizing fasciitis caused by Vibrio vulnificus and Staphylococcus aureus is a surgical emergency. Vibrio vulnificus infection progresses more rapidly and the clinical characteristics are more fulminant than either methicillin-resistant Staphylococcus aureus or methicillin-sensitive Staphylococcus aureus infection.


HTLV-1 Tax Induces Formation of the Active Macromolecular IKK Complex by Generating Lys63- and Met1-Linked Hybrid Polyubiquitin Chains.
The Tax protein of human T-cell leukemia virus type 1 (HTLV-1) is crucial for the development of adult T-cell leukemia (ATL), a highly malignant CD4+ T cell neoplasm. Among the multiple aberrant Tax-induced effects on cellular processes, persistent activation of transcription factor NF-κB, which is activated only transiently upon physiological stimulation, is essential for leukemogenesis. We and others have shown that Tax induces activation of the IκB kinase (IKK) complex, which is a critical step in NF-κB activation, by generating Lys63-linked polyubiquitin chains. However, the molecular mechanism underlying Tax-induced IKK activation is controversial and not fully understood. Here, we demonstrate that Tax recruits linear (Met1-linked) ubiquitin chain assembly complex (LUBAC) to the IKK complex and that Tax fails to induce IKK activation in cells that lack LUBAC activity. Mass spectrometric analyses revealed that both Lys63-linked and Met1-linked polyubiquitin chains are associated with the IKK complex. Furthermore, treatment of the IKK-associated polyubiquitin chains with Met1-linked-chain-specific deubiquitinase (OTULIN) resulted in the reduction of high molecular weight polyubiquitin chains and the generation of short Lys63-linked ubiquitin chains, indicating that Tax can induce the generation of Lys63- and Met1-linked hybrid polyubiquitin chains. We also demonstrate that Tax induces formation of the active macromolecular IKK complex and that the blocking of Tax-induced polyubiquitin chain synthesis inhibited formation of the macromolecular complex. Taken together, these results lead us to propose a novel model in which the hybrid-chain-dependent oligomerization of the IKK complex triggered by Tax leads to trans-autophosphorylation-mediated IKK activation.


Women's well-being improves after missed miscarriage with more active support and application of Swanson's Caring Theory.
The purpose of this study was to provide better organization and more efficient use of resources within the health care system in order to identify women with nonviable pregnancy earlier in their gestation terms and also to identify those women who experience severe grief reaction after the miscarriage. The proposed solution is to offer an appointment with a gynecologist during regular office hours after consultation with the patient's midwife to women experiencing symptoms and who are concerned with the viability of their pregnancy. Unnecessary contact with the emergency room by the patients would be reduced as a result of this improvement in organization. The aim of the study was to give the women experiencing missed miscarriage an increased sense of well-being by applying Swanson's Caring Theory to their recovery, in addition to the better organization and more efficient use of resources. Both the original study from 2002 to 2003 and the later study from 2004 to 2005 applied Swanson's Caring Theory in the follow-up care management of the women, but only the later study was influenced by the changes made in the health care system. In the past, diagnosis of missed miscarriage was delayed because women experiencing minor symptoms were not highly prioritized in the health care system. More active support was introduced in order to get the proper information to the patient throughout the health care system. The size of the original study database was n = 43, compared with the later study database, which was n = 56. All of the women answered the Perinatal Grief Scale (PGS) questions twice, 1 month and 4 months after their diagnosis. Some additional questions about their circumstances unrelated to the PGS were also mailed to the women 4 months after their diagnosis. As a result of the more active support, women felt that they received professional care when they needed it most. The patients were satisfied that they were treated as if they were suffering from normal grief. The group score above the limits for deep grief 4 months after diagnosis was significantly lowered. The chances of receiving their diagnosis at an appointment during office hours increased (odds ratio 3.38). Sick leave time of more than a week was reduced from 44% in the original study to 22% in the later study.


Upscaling tropical restoration to deliver environmental benefits and socially equitable outcomes.
The UN Decade on Ecosystem Restoration offers immense potential to return hundreds of millions of hectares of degraded tropical landscapes to functioning ecosystems. Well-designed restoration can tackle multiple Sustainable Development Goals, driving synergistic benefits for biodiversity, ecosystem services, agricultural and timber production, and local livelihoods at large spatial scales. To deliver on this potential, restoration efforts must recognise and reduce trade-offs among objectives, and minimize competition with food production and conservation of native ecosystems. Restoration initiatives also need to confront core environmental challenges of climate change and inappropriate planting in savanna biomes, be robustly funded over the long term, and address issues of poor governance, inadequate land tenure, and socio-cultural disparities in benefits and costs. Tackling these issues using the landscape approach is vital to realising the potential for restoration to break the cycle of land degradation and poverty, and deliver on its core environmental and social promises.


Comparative benefits and harms of second generation antidepressants and cognitive behavioral therapies in initial treatment of major depressive disorder: systematic review and meta-analysis.
What are the benefits and harms of second generation antidepressants and cognitive behavioral therapies (CBTs) in the initial treatment of a current episode of major depressive disorder in adults? This was a systematic review including qualitative assessment and meta-analyses using random and fixed effects models. Medline, Embase, the Cochrane Library, the Allied and Complementary Medicine Database, PsycINFO, and the Cumulative Index to Nursing and Allied Health Literature were searched from January 1990 through January 2015. The 11 randomized controlled trials included compared a second generation antidepressant CBT. Ten trials compared antidepressant monotherapy with CBT alone; three compared antidepressant monotherapy with antidepressant plus CBT. Meta-analyses found no statistically significant difference in effectiveness between second generation antidepressants and CBT for response (risk ratio 0.91, 0.77 to 1.07), remission (0.98, 0.73 to 1.32), or change in 17 item Hamilton Rating Scale for Depression score (weighted mean difference, -0.38, -2.87 to 2.10). Similarly, no significant differences were found in rates of overall study discontinuation (risk ratio 0.90, 0.49 to 1.65) or discontinuation attributable to lack of efficacy (0.40, 0.05 to 2.91). Although more patients treated with a second generation antidepressant than receiving CBT withdrew from studies because of adverse events, the difference was not statistically significant (risk ratio 3.29, 0.42 to 25.72). No conclusions could be drawn about other outcomes because of lack of evidence. Results should be interpreted cautiously given the low strength of evidence for most outcomes. The scope of this review was limited to trials that enrolled adult patients with major depressive disorder and compared a second generation antidepressant with CBT, and many of the included trials had methodological shortcomings that may limit confidence in some of the findings. Second generation antidepressants and CBT have evidence bases of benefits and harms in major depressive disorder. Available evidence suggests no difference in treatment effects of second generation antidepressants and CBT, either alone or in combination, although small numbers may preclude detection of small but clinically meaningful differences. Funding, competing interests, data sharing This project was funded under contract from the Agency for Healthcare Research and Quality by the RTI-UNC Evidence-based Practice Center. Detailed methods and additional information are available in the full report, available at http://effectivehealthcare.ahrq.gov/.


Next-generation sequencing reveals the biological significance of the N(2),3-ethenoguanine lesion in vivo.
Etheno DNA adducts are a prevalent type of DNA damage caused by vinyl chloride (VC) exposure and oxidative stress. Etheno adducts are mutagenic and may contribute to the initiation of several pathologies; thus, elucidating the pathways by which they induce cellular transformation is critical. Although N(2),3-ethenoguanine (N(2),3-εG) is the most abundant etheno adduct, its biological consequences have not been well characterized in cells due to its labile glycosidic bond. Here, a stabilized 2'-fluoro-2'-deoxyribose analog of N(2),3-εG was used to quantify directly its genotoxicity and mutagenicity. A multiplex method involving next-generation sequencing enabled a large-scale in vivo analysis, in which both N(2),3-εG and its isomer 1,N(2)-ethenoguanine (1,N(2)-εG) were evaluated in various repair and replication backgrounds. We found that N(2),3-εG potently induces G to A transitions, the same mutation previously observed in VC-associated tumors. By contrast, 1,N(2)-εG induces various substitutions and frameshifts. We also found that N(2),3-εG is the only etheno lesion that cannot be repaired by AlkB, which partially explains its persistence. Both εG lesions are strong replication blocks and DinB, a translesion polymerase, facilitates the mutagenic bypass of both lesions. Collectively, our results indicate that N(2),3-εG is a biologically important lesion and may have a functional role in VC-induced or inflammation-driven carcinogenesis.


Psychodidactic determinants of the development of children of preschool age.
The article analyzes the state and trends of the problem of the development of children of preschool age. As a research task, the authors present the results of questionnaires of parents of preschool-age children. The foreign experience of pre-school education is described. The psychodidactic contradictions and perspectives of the development of preschool-age children are outlined. According to scientists, modern preschool education in the context of globalization changes should be reoriented to the development of the child's personality as the main resource, which determines the progressive movement of society. A modern high-tech world puts the child in front of the child's demands of the level of his education, development and upbringing. Consequently, the study of the psychodidactic determinants of the development of a child of preschool age, which provides a complete psycho-physical and personal development and psychological readiness for studying at school, is relevant. The article analyzes the indicators of the cognitive and extracognitive development of the child, in accordance with the most well-known systems of early education, and determines the factors of influence (determinants) of the mental development of a child of preschool age. For conducting scientific researches, the authors used methods of analysis of normative and scientific literature, statistical, system analysis and generalizations, results of their own empirical studies, materials of questionnaires, method of factor-criterion modeling of spheres of mental development of a child, created on the basis of qualimetric evaluation, etc. It has been established that pre-school education should ensure the harmonious development of the child's personality, his physical and mental health, to form mechanisms for social adaptation and creative implementation in a living environment in the community of children and adults through the creation of early education technologies that include a balanced mix of development cognitive and noncognitive spheres of the child. In order to achieve the tasks, innovative pedagogical technologies such as modern systems of early development are widely used, the main purpose of which, according to their authors, is the development of cognitive, emotional and noncognitive abilities of the child. Having examined the main (most popular) early-developmental systems of skills formation in cognitive and non-cognitive spheres, one can conclude that there is a need for comprehensive (systemic) approaches to the development of a child's early developmental technology, since none of the systems considered alone can provide a harmonious balanced development of preschoolers in cognitive (ability to read, write, read, analyze text, perform creative tasks, etc.) and non-cognitive (social, emotional, ability to play storyline games, with independence, etc.) spheres. After all, if loads are lacking, then mental development stops, and excessive tension and absolute comfort lead to a stop of the development of the psyche, self-awareness, therefore, only a balanced approach will allow to get a positive result.


Characterising a healthy adult with a rare HAO1 knockout to support a therapeutic strategy for primary hyperoxaluria.
By sequencing autozygous human populations, we identified a healthy adult woman with lifelong complete knockout of HAO1 (expected ~1 in 30 million outbred people). HAO1 (glycolate oxidase) silencing is the mechanism of lumasiran, an investigational RNA interference therapeutic for primary hyperoxaluria type 1. Her plasma glycolate levels were 12 times, and urinary glycolate 6 times, the upper limit of normal observed in healthy reference individuals (n = 67). Plasma metabolomics and lipidomics (1871 biochemicals) revealed 18 markedly elevated biochemicals (>5 sd outliers versus n = 25 controls) suggesting additional HAO1 effects. Comparison with lumasiran preclinical and clinical trial data suggested she has <2% residual glycolate oxidase activity. Cell line p.Leu333SerfsTer4 expression showed markedly reduced HAO1 protein levels and cellular protein mis-localisation. In this woman, lifelong HAO1 knockout is safe and without clinical phenotype, de-risking a therapeutic approach and informing therapeutic mechanisms. Unlocking evidence from the diversity of human genetic variation can facilitate drug development.


Clostridium difficile Infections amongst Patients with Haematological Malignancies: A Data Linkage Study.
Identify risk factors for Clostridium difficile infection (CDI) and assess CDI outcomes among Australian patients with a haematological malignancy. A retrospective cohort study involving all patients admitted to hospitals in Western Australia with a haematological malignancy from July 2011 to June 2012. Hospital admission data were linked with all hospital investigated CDI case data. Potential risk factors were assessed by logistic regression. The risk of death within 60 and 90 days of CDI was assessed by Cox Proportional Hazards regression. There were 2085 patients of whom 65 had at least one CDI. Twenty percent of CDI cases were either community-acquired, indeterminate source or had only single-day admissions in the 28 days prior to CDI. Using logistic regression, having acute lymphocytic leukaemia, neutropenia and having had bacterial pneumonia or another bacterial infection were associated with CDI. CDI was associated with an increased risk of death within 60 and 90 days post CDI, but only two deaths had CDI recorded as an antecedent factor. Ribotyping information was available for 33 of the 65 CDIs. There were 19 different ribotypes identified. Neutropenia was strongly associated with CDI. While having CDI is a risk factor for death, in many cases it may not be a direct contributor to death but may reflect patients having higher morbidity. A wide variety of C. difficile ribotypes were found and community-acquired infection may be under-estimated in these patients.


Dyskeratosis congenita. Report of a case with literature review.
Leukoplakic lesion is not uncommon in clinical practice but its occurrence as a component of a syndrome is rare. Dyskeratosis congenita is a rare genodermatosis, which is characterized by triad of skin pigmentation, nail dystrophy and leukoplakic lesion in the oral cavity. It is important for dentists to now about Dyskeratosis Congenita because these leukoplakic lesions can spontaneously undergo malignant transformation. Majority of cases have been reported in dermatology or pediatrics literature whereas only few reports have appeared in dental literature. The purpose of reporting this case with review of recent literature is to create better awareness among dentists about the multisystem manifestations and oro-dental abnormalities of this fatal condition that can aid clinicians in early diagnosis. A case of 26 year old male is reported with details of differential diagnosis, importance of hematological investigations, periodic biopsies and current concepts in management of such patients are briefly reviewed.


Extended viewing angle holographic display system with tilted SLMs in a circular configuration.
This paper presents an extended viewing angle holographic display for reconstruction of real world objects in which the capture and display systems are decoupled. This is achieved by employing multiple tilted spatial light modulators (SLMs) arranged in a circular configuration. In order to prove the proper reconstruction and visual perception of holographic images the Wigner distribution function is employed. We describe both the capture system using a single static camera with a rotating object and a holographic display utilizing six tilted SLMs. The experimental results based on the reconstruction of computer generated and real world scenes are presented. The coherent noise removal procedure is described and implemented. The experiments prove the possibility to view images reconstructed in the display binocularly and with good quality.


Procedure volume of gastric cancer resections versus 5-year survival.
We used nationwide, population-based data to examine associations between hospital and surgeon volumes of gastric cancer resections and their patients' short-term and long-term survival likelihood. The study uses 1997-1999 inpatient claims data from Taiwan's National Health Insurance linked to "cause of death" data for 1997-2004. The total cohort of 6909 gastric cancer resection patients were categorized by their surgeon's/hospital's procedure volume, and examined for differences in 6-month mortality and 5-year mortality (post 6 months), by procedure volume, using Cox proportional hazard regressions, adjusting for surgeon, hospital and patient characteristics. We hypothesized that surgeons' case volume and age but not hospital volume will predict short-term and long-term survival. Adjusted estimates show that increasing surgeon volume predicts better 6-month survival (adjusted mortality hazard ratio = 1.3 for low-volume surgeons relative to very high-volume surgeons; p < 0.01) and 5-year survival (adjusted mortality hazard ratios = 1.3; p < 0.001 for low-volume; 1.2 with p < 0.01 for medium volume) and increasing surgeon's age (adjusted hazards ratio = 1.4 for age < 41 years relative to 41-50 years; p < or = 0.001; 0.8 for > or = 51 years relative to 41-50 years; p < 0.05). In hospital volume regressions, surgeon's age is a consistent and significant predictor, not hospital volume. Findings suggest a key role of experience in surgical skill and sensitivity for early stage diagnosis in gastric cancer survival. Although a key study limitation is the lack of cancer stage data, the pattern of findings suggests that experienced surgeons have relatively better survival outcomes among gastric cancer patients.


Periradicular curettage.
Periradicular curettage is a part of the treatment procedure of periradicular surgery. Its main purpose is to remove pathological periradicular tissues for visibility and accessibility to facilitate the treatment of the apical root canal system, or sometimes for the removal of harmful foreign materials present in the periradicular area. Inflammatory periradicular lesions (granuloma and cysts) are the responses of the periradicular tissues to irritants from the root canal and not from the periradicular area unless medicaments and/or filling materials have been forced through the apical foramina or perforations into the periodontium. Histologically, the inflammatory periradicular lesion is similar to healing granulation tissue, which is composed of cells which have natural and specific immunological defence capability and cooperate by means of cytokines to amplify the protective mechanisms of the host. Accordingly, it is not necessary to completely curette out all the inflamed periradicular tissues during surgery, since this granulation-like tissue will be incorporated into the new granulation tissue as part of the healing process. To control the source of irritants in the root canal is far more important than to remove all periradicular tissues affected by the irritants. The successful removal of all irritants from the root canal system results in resolution of pulpally induced periradicular lesions. In the case where the periradicular lesion is caused by endodontic instruments or cytotoxic filling materials placed in the periradicular tissues, removal of these foreign objects is required for resolution of the lesion.


Rapid, efficient, and simple motor neuron differentiation from human pluripotent stem cells.
Human pluripotent stem cells (hPSCs) are being applied in regenerative medicine and for the in vitro modeling of human intractable disorders. In particular, neural cells derived from disease-specific human induced pluripotent stem cells (hiPSCs) established from patients with neurological disorders have been used as in vitro disease models to recapitulate in vivo pathogenesis because neural cells cannot be usually obtained from patients themselves. In this study, we established a rapid, efficient, and simple method for efficiently deriving motor neurons from hPSCs that is useful for pathophysiological analysis and the development of drugs to treat motor neuron diseases. Treatment with GSK3β inhibitors during the initial phase of differentiation in combination with dual SMAD inhibition was sufficient to induce PAX6 (+) and SOX1 (+) neural progenitors within 1 week, and subsequent treatment with retinoic acid (RA) and purmorphamine, which activates sonic hedgehog (SHH) signaling, resulted in the highly efficient induction of HB9(+) and ISL-1(+) motor neurons within 2 weeks. After 4 weeks of monolayer differentiation in motor neuron maturation medium, hPSC-derived motor neurons were shown to mature, displaying larger somas and clearer staining for the mature motor neuron marker choline acetyltransferase (ChAT). Moreover, hPSC-derived motor neurons were able to form neuromuscular junctions with human myotubes in vitro and induced acetylcholine receptor (AChR) clustering, as detected by Alexa 555-conjugated α-Bungarotoxin (α-BTX), suggesting that these hPSC-derived motor neurons formed functional contacts with skeletal muscles. This differentiation system is simple and is reproducible in several hiPSC clones, thereby minimizing clonal variation among hPSC clones. We also established a system for visualizing motor neurons with a lentiviral reporter for HB9 (HB9 (e438) ::Venus). The specificity of this reporter was confirmed through immunocytochemistry and quantitative RT-PCR analysis of high-positive fractions obtained via fluorescence-activated cell sorting (FACS), suggesting its applicability for motor neuron-specific analysis. Our motor neuron differentiation system and lentivirus-based reporter system for motor neurons facilitate the analysis of disease-specific hiPSCs for motor neuron diseases.


Reconfigurable optical manipulation by phase change material waveguides.
Optical manipulation by dielectric waveguides enables the transportation of particles and biomolecules beyond diffraction limits. However, traditional dielectric waveguides could only transport objects in the forward direction which does not fulfill the requirements of the next generation lab-on-chip system where the integrated manipulation system should be much more flexible and multifunctional. In this work, bidirectional transportation of objects on the nanoscale is demonstrated on a rectangular waveguide made of the phase change material Ge2Sb2Te5 (GST) by numerical simulations. Either continuous pushing forces or pulling forces are generated on the trapped particles when the GST is in the amorphous or crystalline phase. With the technique of a femtosecond laser induced phase transition on the GST, we further proposed a reconfigurable optical trap array on the same waveguide. This work demonstrates GST waveguide's potential of achieving multifunctional manipulation of multiple objects on the nanoscale with plausible optical setups.


Cocaine-exposed preterm neonates show behavioral and hormonal differences.
Prematurity has been associated with prenatal cocaine exposure, but most studies on the behavioral effects of prenatal cocaine exposure have been restricted to full-term infant samples. The current study focused on behavioral and hormonal responses in preterm cocaine-exposed infants compared with a cohort of non-cocaine-exposed infants of similar gestational age. A comparison between 30 cocaine-exposed and 30 non-cocaine-exposed preterm neonates suggested that the cocaine-exposed neonates were born to mothers who had higher parity and more obstetric complications. In addition, mothers of cocaine-exposed preterm neonates visited, touched, held, and fed their infants less frequently than mothers of nonexposed infants. The cocaine-exposed infants had smaller head circumferences at birth, spent more time in the neonatal intensive care unit, and had a greater incidence of periventricular-intraventricular hemorrhages. They also had inferior Brazelton cluster scores, including lower state regulation and range-of-state scores, and greater depression. During sleep-wake behavior observations, they showed difficulty maintaining alert states and self-regulating their behavior, and they spent more time in indeterminate sleep and had decreased periods of quiet sleep and increased levels of agitated behavior, including tremulousness, mouthing, multiple limb movements, and clenched fists. Finally, higher urinary norepinephrine, dopamine, and cortisol levels and lower plasma insulin levels were noted in the cocaine-exposed preterm neonates. These findings highlight the need for follow-up assessments and early intervention.


Fluorescent magnetic nanocrystals by sequential addition of reagents in a one-pot reaction: a simple preparation for multifunctional nanostructures.
Core-shell nanostructures consisting of FePt magnetic nanoparticles as the core and semiconducting chalcogenides as the shell were synthesized by a series of reactions in a one-pot procedure. Adding Cd(acac)2 as the cadmium precursor to a reaction mixture containing FePt nanoparticles afforded FePt@CdO core-shell intermediates. The subsequent addition of chalcogens yielded FePt@CdX core-shell nanocrystals (where X was S or Se). The reverse sequence of addition, i.e., adding X before Cd, resulted in spongelike nanostructures because the chalcogens readily formed nanowires in the solution. Transmission electron microscopy, energy-dispersive X-ray spectrometry, selected area electron diffraction, fluorescence spectroscopy, and SQUID were used to characterize the nanostructures. These core-shell nanostructures displayed superparamagnetism at room temperature and exhibited fluorescence with quantum yields of 2.3-9.7%. The flexibility in the sequence of addition of reagents, combined with the compatibility of the lattices of the different materials, provides a powerful yet convenient strategy for generating sophisticated, multifunctional nanostructures.


Muscle cramp in Machado-Joseph disease: altered motor axonal excitability properties and mexiletine treatment.
Machado-Joseph disease is one of the most common hereditary spinocerebellar degenerative disorders with a wide range of clinical manifestations. Pathology studies have shown mild to moderate loss of anterior horn cells and, in terms of spinal pathology, Machado-Joseph disease is regarded as a type of lower motoneuron disease. Muscle cramps are often associated with lower motoneuron disorders, but features of cramps in Machado-Joseph disease patients have never been studied. We investigated the incidence and nature of muscle cramps in Machado-Joseph disease patients, the excitability properties of motor axons [strength-duration time constant (tau(SD)), threshold electrotonus, refractoriness and supernormality] using threshold tracking and the effects of mexiletine hydrochloride on those cramps. Of 20 consecutive patients, 16 (80%) had frequent, severe muscle cramps in the legs, trunk or arms that disturbed their daily activities. The frequency of pathological muscle cramps was similar to that for patients with amyotrophic lateral sclerosis (68%) and higher than those for patients with spinal muscular atrophy (33%) or peripheral axonal neuropathy (24%). Threshold-tracking studies showed that tau(SD), which in part reflects Na(+) conductance at the resting membrane potential, was significantly greater in the Machado-Joseph disease patients than in normal subjects; severe muscle cramps were associated with a longer tau(SD). Threshold electrotonus, refractoriness and supernormality were not significantly different between Machado-Joseph disease patients and normal subjects. Eight Machado-Joseph disease patients with severe cramps, who received mexiletine treatment, experienced nearly complete relief with a partial normalization of tau(SD) (P = 0.08). Muscle cramps are a very frequent and disabling factor in Machado-Joseph disease. Pathological muscle cramps responded well to mexiletine treatment, and this is consistent with the hypothesis that they are caused by an increase in persistent Na(+) conductance, possibly associated with axonal regeneration or collateral sprouting.


Treatment for depression comorbid with dementia.
Depression is a common comorbidity in dementia. Randomised controlled studies of antidepressants do not show a significant improvement in depressive symptoms in patients with comorbid dementia and are known to lead to an increase in side effects. However, there are relatively few studies of depression in dementia, and drawing firm conclusions about the use of antidepressants is limited by the amount of data available. Furthermore, it is unclear whether data can be extrapolated from similar populations (eg, those with late-life depression) to inform pharmacotherapy in this patient group. Given the lack of effectiveness and risk of side effects associated with pharmacological treatments, psychological interventions may offer important therapeutic benefits. There is evidence for the effectiveness of individual psychological therapy, and further research will establish which psychological approach is the most effective. Some studies have shown an improvement in depressive symptoms using structured sleep hygiene programmes, exercise, arts interventions and music therapy. These studies are hampered by small data sets, and the benefits to individuals may not be well captured by standard outcome measures. At present, the best evidence for arts-based approaches is in music therapy. Depression with comorbid dementia responds well to electroconvulsive therapy and this is a useful treatment modality for those with severe or life-threatening depressive symptoms. Alternative neurostimulation techniques such as transcranial magnetic stimulation are not widely used at present and further research is needed before they can be a more widely used treatment modality.


[The anti-HIV activity of glycyrrhizic acid penta-O-nicotinate].
The anti-HIV activity of niglizin (penta-O-nicotinate of glycyrrhizic acid) and of its combinations was studied in the culture of infected MT-4 cells and in respect to the recombinant reverse HIV-1 transcriptase. Niglizin was shown to suppress effectively the HIV replication and to be a noncompetitive inhibitor of reverse transcriptase. Research of a combined anti-HIV action of niglizin and of azidothimidine (AZT) demonstrated that the preparations, when used at ratios of 1:20, 1:50, 1:200 and 1:2000, suppressed the synergetic effect both in the cell culture and in the recombinant reverse HIV transcriptase. A study of the antiviral activity of niglizin and of its joint use with AZT in respect to the AZT-resistant HIV-1 mutant showed niglizin to be more effective (ID50 = 0.134 microM) versus the "wild" strain. (ID50 = 9.64 micriM); whereas, its combined use AZT:niglizin = 1:100 displayed synergism (FIC = 0.553). The efficiency of the combined AZT/niglizin anti-HIV effect both in respect to the "wild" strain and to the AZT-resistant mutant confirms that such combinations are promising for the treatment of HIV infection.


Clinical and pharmacological studies on liver diseases treated with Kampo herbal medicine.
Hepatitis C virus (HCV) infection frequently causes chronic hepatitis, which is linked to the development of liver cirrhosis and hepatocellular carcinoma. Most physicians who practice Kampo medicine in Japan have observed that Kampo medicine can be as effective as interferon therapy in the treatment of chronic hepatitis C. In the present study, to evaluate the effect of Kampo medicine on chronic hepatitis C, clinical treatment was assessed in short-term and long-term study, and it was shown that ninjin-yoei-to (Formula ginseng compositae: TJ-108) was very effective. Therefore, to find the most active herbal component of TJ-108 in the treatment of HCV, Citrus Unshiu Peel, Schisandra Fruit, and Polygala Root, which are specific to TJ-108, were screened using an in vitro HCV infection model. Among the three herbs, Schisandra Fruit was found to be most active. In the next step, Gomisin A, an active component of Schisandra Fruit, was studied using an in vitro model with MOLT-4 cells and an animal model of immunologically induced acute hepatic failures. It is concluded that the therapeutic effect of TJ-108 on chronic hepatitis C is from the inhibitory effect on HCV infection, and also from the protective effect on immunological hepatopathy of Schisandra Fruit and its lignan component, Gomisin A.


Predictors of a long length of stay in the emergency department for older people.
Dedicated geriatric models of care are becoming more prevalent due to the complexity of, and increase in, acute healthcare presentations for older patients. For older people, a long stay in the emergency department (ED) may reflect the complexity of their presentation, or deficiencies in systems that manage these complexities. To identify predictors of a long ED length of stay (LLoS) for patients ≥65 years old. Linked hospital information systems data from a large, public Australian ED were analysed in this retrospective cohort study. LLoS was defined as the 75th percentile (617 min). Multivariate regression identified LLoS predictors for admissions and discharges separately. Of 16 791 ED presentations made by older people, 4192 experienced a LLoS; 55% were admitted. Increasing age was associated with an increasing ED LoS. Factors most predictive of LLoS for both admitted and discharged patients included: investigations (both pathology and imaging), less urgent Australasian triage scale categories and after-hours arrival. Ambulance arrival did not increase the risk of a LLoS for patients eventually admitted, but conferred nearly a twofold increased risk for a LLoS for discharged older persons (adjusted odds ratios = 1.9; 95% confidence interval 1.5-2.4). This study assists clinicians and decision-makers to identify reasons why older persons have a LLoS, whether admitted or discharged. Interventions to streamline care for older patients arriving after-hours and who require imaging and pathology are required. LoS targets should consider age distribution. The use of ED LoS as a quality of care indicator should be assessed for admissions and discharges, separately.


Genomic exploration of individual giant ocean viruses.
Viruses are major pathogens in all biological systems. Virus propagation and downstream analysis remains a challenge, particularly in the ocean where the majority of their microbial hosts remain recalcitrant to current culturing techniques. We used a cultivation-independent approach to isolate and sequence individual viruses. The protocol uses high-speed fluorescence-activated virus sorting flow cytometry, multiple displacement amplification (MDA), and downstream genomic sequencing. We focused on 'giant viruses' that are readily distinguishable by flow cytometry. From a single-milliliter sample of seawater collected from off the dock at Boothbay Harbor, ME, USA, we sorted almost 700 single virus particles, and subsequently focused on a detailed genome analysis of 12. A wide diversity of viruses was identified that included Iridoviridae, extended Mimiviridae and even a taxonomically novel (unresolved) giant virus. We discovered a viral metacaspase homolog in one of our sorted virus particles and discussed its implications in rewiring host metabolism to enhance infection. In addition, we demonstrated that viral metacaspases are widespread in the ocean. We also discovered a virus that contains both a reverse transcriptase and a transposase; although highly speculative, we suggest such a genetic complement would potentially allow this virus to exploit a latency propagation mechanism. Application of single virus genomics provides a powerful opportunity to circumvent cultivation of viruses, moving directly to genomic investigation of naturally occurring viruses, with the assurance that the sequence data is virus-specific, non-chimeric and contains no cellular contamination.


New-onset seizures in an elderly hospitalized population.
We studied 80 hospitalized patients over 60 years old with either new-onset or newly diagnosed seizures that were generalized tonic-clonic in approximately half the cases and partial with or without secondary generalization in the other half. The etiology of seizures was acute symptomatic in 33 (41%), remote symptomatic in 32 (40%), progressive encephalopathy in nine (11%), and idiopathic in six (8%). Convulsive status epilepticus occurred in five cases (6%). The most common single cause of seizures was infarction or hemorrhage (54%). Morbidity and mortality were highest in the acute symptomatic group (p < 0.03). Nine (11%) of the subjects died within 3 months of admission, including two of the five with status epilepticus. Of the patients with acute symptomatic etiologies, 21% died compared with 6% of those in the remote symptomatic group. New neurologic deficits were present in eight (11%) of the 71 who survived, including five acute symptomatic, one remote symptomatic, and two progressive encephalopathy cases. No patient with idiopathic seizures died or had new neurologic deficits. We conclude that seizures in the elderly requiring hospitalization occur mainly with acute and remote symptomatic neurologic insults and are associated with a significant morbidity and mortality. In the absence of any associated neurologic insults, the morbidity is low.


Glycoprotein B of equine herpesvirus type 1 has two recognition sites for subtilisin-like proteases that are cleaved by furin.
Glycoprotein B (gB) of equine herpesvirus type 1 (EHV-1) is predicted to be cleaved by furin in a fashion similar to that of related herpesviruses. To investigate the contribution of furin-mediated gB cleavage to EHV-1 growth, canonical furin cleavage sites were mutated. Western blot analysis of mutated EHV-1 gB showed that it was cleaved at two positions, 518RRRR521 and 544RLHK547, and that the 28 aa between the two sites were removed after cleavage. Treating infected cells with either convertase or furin inhibitors reduced gB cleavage efficiency. Further, removal of the first furin recognition motif did not affect in vitro growth of EHV-1, while mutation of the second motif greatly affected virus growth. In addition, a second possible signal peptide cleavage site was identified for EHV-1 gB between residues 98 and 99, which was 13 aa downstream of that previously identified.


Pathogenicity of new BEST1 variants identified in Italian patients with best vitelliform macular dystrophy assessed by computational structural biology.
Best vitelliform macular dystrophy (BVMD) is an autosomal dominant macular degeneration. The typical central yellowish yolk-like lesion usually appears in childhood and gradually worsens. Most cases are caused by variants in the BEST1 gene which encodes bestrophin-1, an integral membrane protein found primarily in the retinal pigment epithelium. Here we describe the spectrum of BEST1 variants identified in a cohort of 57 Italian patients analyzed by Sanger sequencing. In 13 cases, the study also included segregation analysis in affected and unaffected relatives. We used molecular mechanics to calculate two quantitative parameters related to calcium-activated chloride channel (CaCC composed of 5 BEST1 subunits) stability and calcium-dependent activation and related them to the potential pathogenicity of individual missense variants detected in the probands. Thirty-six out of 57 probands (63% positivity) and 16 out of 18 relatives proved positive to genetic testing. Family study confirmed the variable penetrance and expressivity of the disease. Six of the 27 genetic variants discovered were novel: p.(Val9Gly), p.(Ser108Arg), p.(Asn179Asp), p.(Trp182Arg), p.(Glu292Gln) and p.(Asn296Lys). All BEST1 variants were assessed in silico for potential pathogenicity. Our computational structural biology approach based on 3D model structure of the CaCC showed that individual amino acid replacements may affect channel shape, stability, activation, gating, selectivity and throughput, and possibly also other features, depending on where the individual mutated amino acid residues are located in the tertiary structure of BEST1. Statistically significant correlations between mean logMAR best-corrected visual acuity (BCVA), age and modulus of computed BEST1 dimerization energies, which reflect variations in the in CaCC stability due to amino acid changes, permitted us to assess the pathogenicity of individual BEST1 variants. Using this computational approach, we designed a method for estimating BCVA progression in patients with BEST1 variants.


Mucopolysaccharidosis VI in cats - clarification regarding genetic testing.
The release of new DNA-based diagnostic tools has increased tremendously in companion animals. Over 70 different DNA variants are now known for the cat, including DNA variants in disease-associated genes and genes causing aesthetically interesting traits. The impact genetic tests have on animal breeding and health management is significant because of the ability to control the breeding of domestic cats, especially breed cats. If used properly, genetic testing can prevent the production of diseased animals, causing the reduction of the frequency of the causal variant in the population, and, potentially, the eventual eradication of the disease. However, testing of some identified DNA variants may be unwarranted and cause undo strife within the cat breeding community and unnecessary reduction of gene pools and availability of breeding animals. Testing for mucopolysaccharidosis Type VI (MPS VI) in cats, specifically the genetic testing of the L476P (c.1427T>C) and the D520N (c.1558G>A) variants in arylsulfatase B (ARSB), has come under scrutiny. No health problems are associated with the D520N (c.1558G>A) variant, however, breeders that obtain positive results for this variant are speculating as to possible correlation with health concerns. Birman cats already have a markedly reduced gene pool and have a high frequency of the MPS VI D520N variant. Further reduction of the gene pool by eliminating cats that are heterozygous or homozygous for only the MPS VI D520N variant could lead to more inbreeding depression effects on the breed population. Herein is debated the genetic testing of the MPS VI D520N variant in cats. Surveys from different laboratories suggest the L476P (c.1427T>C) disease-associated variant should be monitored in the cat breed populations, particularly breeds with Siamese derivations and outcrosses. However, the D520N has no evidence of association with disease in cats and testing is not recommended in the absence of L476P genotyping. Selection against the D520N is not warranted in cat populations. More rigorous guidelines may be required to support the genetic testing of DNA variants in all animal species.


Increased affluence explains the emergence of ascetic wisdoms and moralizing religions.
Between roughly 500 BCE and 300 BCE, three distinct regions, the Yangtze and Yellow River Valleys, the Eastern Mediterranean, and the Ganges Valley, saw the emergence of highly similar religious traditions with an unprecedented emphasis on self-discipline and asceticism and with "otherworldly," often moralizing, doctrines, including Buddhism, Jainism, Brahmanism, Daoism, Second Temple Judaism, and Stoicism, with later offshoots, such as Christianity, Manichaeism, and Islam. This cultural convergence, often called the "Axial Age," presents a puzzle: why did this emerge at the same time as distinct moralizing religions, with highly similar features in different civilizations? The puzzle may be solved by quantitative historical evidence that demonstrates an exceptional uptake in energy capture (a proxy for general prosperity) just before the Axial Age in these three regions. Statistical modeling confirms that economic development, not political complexity or population size, accounts for the timing of the Axial Age. We discussed several possible causal pathways, including the development of literacy and urban life, and put forward the idea, inspired by life history theory, that absolute affluence would have impacted human motivation and reward systems, nudging people away from short-term strategies (resource acquisition and coercive interactions) and promoting long-term strategies (self-control techniques and cooperative interactions).


Expression of spoT in Borrelia burgdorferi during serum starvation.
Borrelia burgdorferi, the causative agent of Lyme disease, is transmitted by the tick Ixodes scapularis. A 2.9-kb fragment containing a putative spoT gene was isolated from B. burgdorferi genomic DNA by PCR amplification and cloned into a pBAD24 vector. The cloned gene complemented Escherichia coli mutant strain CF1693, which contains deletions of both the relA and spoT genes. The spoT gene in E. coli encodes a bifunctional enzyme capable of synthesizing and degrading (p)ppGpp, which mediates the stringent response during carbon source starvation. B. burgdorferi has been reported to have a stress response to serum starvation. Thin-layer chromatography was used to detect (p)ppGpp extracted from H(3)(32)PO(4)-labeled B. burgdorferi cells starved for serum in RPMI. B. burgdorferi spoT gene expression was characterized during fatty acid starvation. Northern analysis of spoT revealed detectable message at 2.5 min of starvation in RPMI. Expression of spoT during serum starvation increased approximately 6-fold during the 30 min that starvation conditions were maintained. Further, expression of spoT decreased when serum was added to serum-starved cells. Reverse transcriptase PCR (RT-PCR) was used to detect spoT mRNA from approximately 10(6) cells starved for serum in RPMI for 2.5 to 30 min or incubated in tick saliva for 15 min. Northern blot analysis suggests that spoT transcript was approximately 900 nucleotides in length. RT-PCR amplification of the transcript using several sets of primers confirmed this finding. Additionally, a truncated clone containing only the first 950 bp of the 2,001-bp spoT open reading frame was able to complement E. coli CF1693. The data suggest that B. burgdorferi exhibits a stringent response to serum starvation and during incubation in tick saliva.


Entropy production for complex Langevin equations.
We study irreversible processes for nonlinear oscillators networks described by complex-valued Langevin equations that account for coupling to different thermochemical baths. Dissipation is introduced via non-Hermitian terms in the Hamiltonian of the model. We apply the stochastic thermodynamics formalism to compute explicit expressions for the entropy production rates. We discuss in particular the nonequilibrium steady states of the network characterized by a constant production rate of entropy and flows of energy and particle currents. For two specific examples, a one-dimensional chain and a dimer, numerical calculations are presented. The role of asymmetric coupling among the oscillators on the entropy production is illustrated.


Genomic analysis of the human B-lymphotropic virus (HBLV).
The human B-lymphotropic virus (HBLV) has a double-stranded DNA genome of greater than 110 kilobase pairs, which is consistent with its morphological classification as a herpesvirus. A 9000-base pair cloned probe of HBLV detected specific sequences in DNA and RNA of infected cells but did not hybridize to the genomic DNA of other human herpesviruses including the Epstein-Barr virus, human cytomegalovirus, herpes simplex type I, and varicella-zoster virus. Conversely, while probes obtained from each of the known human herpesvirus readily detected the homologous viral DNA, they did not hybridize to genomic HBLV DNA. This evidence, in addition to serological and morphological distinctions and the biological effects of this virus demonstrate that HBLV is a novel human herpesvirus.


Nonlinear analysis of motor activity shows differences between schizophrenia and depression: a study using Fourier analysis and sample entropy.
The purpose of this study has been to describe motor activity data obtained by using wrist-worn actigraphs in patients with schizophrenia and major depression by the use of linear and non-linear methods of analysis. Different time frames were investigated, i.e., activity counts measured every minute for up to five hours and activity counts made hourly for up to two weeks. The results show that motor activity was lower in the schizophrenic patients and in patients with major depression, compared to controls. Using one minute intervals the depressed patients had a higher standard deviation (SD) compared to both the schizophrenic patients and the controls. The ratio between the root mean square successive differences (RMSSD) and SD was higher in the schizophrenic patients compared to controls. The Fourier analysis of the activity counts measured every minute showed that the relation between variance in the low and the high frequency range was lower in the schizophrenic patients compared to the controls. The sample entropy was higher in the schizophrenic patients compared to controls in the time series from the activity counts made every minute. The main conclusions of the study are that schizophrenic and depressive patients have distinctly different profiles of motor activity and that the results differ according to period length analysed.


Clinical laboratory sciences data transmission: the NPU coding system.
In health care services, technology requires that correct information be duly available to professionals, citizens and authorities, worldwide. Thus, clinical laboratory sciences require standardized electronic exchanges for results of laboratory examinations. The NPU (Nomenclature, Properties and Units) coding system provides a terminology for identification of result values (property values). It is structured according to BIPM, ISO, IUPAC and IFCC recommendations. It uses standard terms for established concepts and structured definitions describing: which part of the universe is examined, which component of relevance in that part, which kind-of-property is relevant. Unit and specifications can be added where relevant [System(spec)-Component(spec); kind-of-property(spec) = ? unit]. The English version of this terminology is freely accessible at http://dior.imt.liu.se/cnpu/ and http://www.labterm.dk, directly or through the IFCC and IUPAC websites. It has been nationally used for more than 10 years in Denmark and Sweden and has been translated into 6 other languages. The NPU coding system provides a terminology for dedicated kinds-of-property following the international recommendations. It fits well in the health network and is freely accessible. Clinical laboratory professionals worldwide will find many advantages in using the NPU coding system, notably with regards to an accreditation process.


An Improved Method for Individual Tracking of Voluntary Wheel Running in Pair-housed Juvenile Mice.
Rodent cages equipped with access to a voluntary running wheel are commonly used to study the effects of aerobic physical activity on physiology and behavior. Notable discoveries in exercise neurobiology, including the key role of brain-derived neurotrophic factor (BDNF) in neural plasticity and cognition, have been made using rodents housed with voluntary running wheels. A major advantage of using home-cage running wheels over treadmills is the elimination of stress potentially associated with forced running. In addition, voluntary wheel running may simulate a more natural running pattern in laboratory mice. Singly housing mice with voluntary running wheels is traditionally employed to obtain exact quantitation of the distance ran; however, social isolation stress is often ignored to obtain precise running distances. Moreover, voluntary exercise studies in adolescent mice must consider the neurodevelopmental implications of isolation stress. In this protocol, we wean 21-day-old mouse pups directly into running wheel-equipped cages and pair-house them to reduce the impact of social isolation and other developmentally specific factors that could adversely affect their behavior or development. Individual running distances are obtained from each mouse in the cage using a radio-frequency identification (RFID) ear tag and a hidden antenna placed directly under the running wheel. We have demonstrated that voluntary running during a specific juvenile-adolescent developmental period can improve hippocampal memory when tested during adolescence ( Ivy et al., 2020 ). Individual exercise tracking of group-housed mice can enable future studies to precisely correlate the amount of exercise with readouts such as cell-specific gene expression, epigenetic mechanisms, serum biomarkers, and behavior, in an intra-individual manner. Graphic abstract: Figure 1.Illustration of the dual RFID and Vital View system for individual mouse running in a pair-housed cage.


A new fetal RHD genotyping test: costs and benefits of mass testing to target antenatal anti-D prophylaxis in England and Wales.
Postnatal and antenatal anti-D prophylaxis have dramatically reduced maternal sensitisations and cases of rhesus disease in babies born to women with RhD negative blood group. Recent scientific advances mean that non-invasive prenatal diagnosis (NIPD), based on the presence of cell-free fetal DNA in maternal plasma, could be used to target prophylaxis on "at risk" pregnancies where the fetus is RhD positive. This paper provides the first assessment of cost-effectiveness of NIPD-targeted prophylaxis compared to current policies. We conducted an economic analysis of NIPD implementation in England and Wales. Two scenarios were considered. Scenario 1 assumed that NIPD will be only used to target antenatal prophylaxis with serology tests continuing to direct post-delivery prophylaxis. In Scenario 2, NIPD would also displace postnatal serology testing if an RhD negative fetus was identified. Costs were estimated from the provider's perspective for both scenarios together with a threshold royalty fee per test. Incremental costs were compared with clinical implications. The basic cost of an NIPD in-house test is £16.25 per sample (excluding royalty fee). The two-dose antenatal prophylaxis policy recommended by NICE is estimated to cost the NHS £3.37 million each year. The estimated threshold royalty fee is £2.18 and £8.83 for Scenarios 1 and 2 respectively. At a £2.00 royalty fee, mass NIPD testing would produce no saving for Scenario 1 and £507,154 per annum for Scenario 2. Incremental cost-effectiveness analysis indicates that, at a test sensitivity of 99.7% and this royalty fee, NIPD testing in Scenario 2 will generate one additional sensitisation for every £9,190 saved. If a single-dose prophylaxis policy were implemented nationally, as recently recommended by NICE, Scenario 2 savings would fall. Currently, NIPD testing to target anti-D prophylaxis is unlikely to be sufficiently cost-effective to warrant its large scale introduction in England and Wales. Only minor savings are calculated and, balanced against this, the predicted increase in maternal sensitisations may be unacceptably high. Reliability of NIPD assays still needs to be demonstrated rigorously in different ethnic minority populations. First trimester testing is unlikely to alter this picture significantly although other emerging technologies may.


Getting to know a stranger--rural nurses' experiences of mentoring: a grounded theory.
Mentoring is often proposed as a solution to the problem of successfully recruiting and retaining nursing staff. The aim of this constructivist grounded theory study was to explore Australian rural nurses' experiences of mentoring. The research design used was reflexive in nature resulting in a substantive, constructivist grounded theory study. A national advertising campaign and snowball sampling were used to recruit nine participants from across Australia. Participants were rural nurses who had experience in mentoring others. Standard grounded theory methods of theoretical sampling, concurrent data collection and analysis using open, axial and theoretical coding and a story line technique to develop the core category and category saturation were used. To cultivate the reflexivity required of a constructivist study, we also incorporated reflective memoing, situational analysis mapping techniques and frame analysis. Data was generated through eleven interviews, email dialogue and shared situational mapping. Cultivating and growing new or novice rural nurses using supportive relationships such as mentoring was found to be an existing, integral part of experienced rural nurses' practice, motivated by living and working in the same communities. Getting to know a stranger is the first part of the process of cultivating and growing another. New or novice rural nurses gain the attention of experienced rural nurses through showing potential or experiencing a critical incidence. The problem of retaining nurses is a global issue. Experienced nurses engaged in clinical practice have the potential to cultivate and grow new or novice nurses-many already do so. Recognising this role and providing opportunities for development will help grow a positive, supportive work environment that nurtures the experienced nurses of tomorrow.


Hydrogen bonding and molecular orientation at the liquid-vapour interface of water.
We determine the molecular structure and orientation at the liquid-vapour interface of water using an electronically coarse grained model constructed to include all long-range electronic responses within Gaussian statistics. The model, fit to the properties of the isolated monomer and dimer, is sufficiently responsive to generate the temperature dependence of the surface tension from ambient conditions to the critical point. Acceptor hydrogen bonds are shown to be preferentially truncated at the free surface under ambient conditions and a related asymmetry in hydrogen bonding preference is identified in bulk water. We speculate that this bonding asymmetry in bulk water is the microscopic origin of the observed surface structure.


The effect of nonmedical factors on variations in the performance of colonoscopy among different health care settings.
Previous published studies have shown significant variations in colonoscopy performance, even when medical factors are taken into account. This study aimed to examine the role of nonmedical factors (ie, embodied in health care system design) as possible contributors to variations in colonoscopy performance. Patient data from a multicenter observational study conducted between 2000 and 2002 in 21 centers in 11 western countries were used. Variability was captured through 2 performance outcomes (diagnostic yield and colonoscopy withdrawal time), jointly studied as dependent variables, using a multilevel 2-equation system. Results showed that open-access systems and high-volume colonoscopy centers were independently associated with a higher likelihood of detecting significant lesions and longer withdrawal durations. Fee for service (FFS) payment was associated with shorter withdrawal durations, and so had an indirect negative impact on the diagnostic yield. Teaching centers exhibited lower detection rates and longer withdrawal times. Our results suggest that gatekeeping colonoscopy is likely to miss patients with significant lesions and that developing specialized colonoscopy units is important to improve performance. Results also suggest that FFS may result in a lower quality of care in colonoscopy practice and highlight the fact that longer withdrawal times do not necessarily indicate higher quality in teaching centers.


The spread of Ras activity triggered by activation of a single dendritic spine.
In neurons, individual dendritic spines isolate N-methyl-d-aspartate (NMDA) receptor-mediated calcium ion (Ca2+) accumulations from the dendrite and other spines. However, the extent to which spines compartmentalize signaling events downstream of Ca2+ influx is not known. We combined two-photon fluorescence lifetime imaging with two-photon glutamate uncaging to image the activity of the small guanosine triphosphatase Ras after NMDA receptor activation at individual spines. Induction of long-term potentiation (LTP) triggered robust Ca2+-dependent Ras activation in single spines that decayed in approximately 5 minutes. Ras activity spread over approximately 10 micrometers of dendrite and invaded neighboring spines by diffusion. The spread of Ras-dependent signaling was necessary for the local regulation of the threshold for LTP induction. Thus, Ca2+-dependent synaptic signals can spread to couple multiple synapses on short stretches of dendrite.


Osteoradionecrosis of the jaws--a current overview--part 1: Physiopathology and risk and predisposing factors.
The aim of this paper is to explore the current theories about definition, classification, incidence and physiopathology of osteoradionecrosis (ORN) of the jaws. Moreover, it is discussed the predisposing and risk factors for the development of osteoradionecrosis based on the literature review. Osteoradionecrosis is one of the most serious oral complications of head and neck cancer treatment. Osteoradionecrosis is a severe delayed radiation-induced injury, characterised by bone tissue necrosis and failure to heal. Osteoradionecrosis either stabilises or gradually worsens and is notoriously difficult to manage. The most widely accepted theory to explain its cause until recently was the theory of hypoxia, hypovascularity and hypocellularity. A new theory for the pathogenesis of osteoradionecrosis was proposed. The clinical presentations of osteoradionecrosis are pain, drainage and fistulation of the mucosa or skin that is related to exposed bone in an area that has been irradiated. The tumour size and location, radiation dose, local trauma, dental extractions, infection, immune defects and malnutrition can predispose its development. A better understanding of risk factors for the development ORN and of the underlying pathophysiology may improve our ability to prevent this complication and help to improve the prognosis for those being treated for osteoradionecrosis.


Eomesodermin is required for mouse trophoblast development and mesoderm formation.
The earliest cell fate decision in the mammalian embryo separates the extra-embryonic trophoblast lineage, which forms the fetal portion of the placenta, from the embryonic cell lineages. The body plan of the embryo proper is established only later at gastrulation, when the pluripotent epiblast gives rise to the germ layers ectoderm, mesoderm and endoderm. Here we show that the T-box gene Eomesodermin performs essential functions in both trophoblast development and gastrulation. Mouse embryos lacking Eomesodermin arrest at the blastocyst stage. Mutant trophoectoderm does not differentiate into trophoblast, indicating that Eomesodermin may be required for the development of trophoblast stem cells. In the embryo proper, Eomesodermin is essential for mesoderm formation. Although the specification of the anterior-posterior axis and the initial response to mesoderm-inducing signals is intact in mutant epiblasts, the prospective mesodermal cells are not recruited into the primitive streak. Our results indicate that Eomesodermin defines a conserved molecular pathway controlling the morphogenetic movements of germ layer formation and has acquired a new function in mammals in the differentiation of trophoblast.


Extrinsic cues aid shape recognition from novel viewpoints.
It has been shown previously that the visual recognition of shape is susceptible to the mismatch between the retinal input and its representation in long-term memory, especially when this mismatch arises from rotations in depth. One possibility is that the visual recognition system deals with such mismatch by a transformation of the input or the representation thereby bringing both into alignment for comparison. In either case, knowing what transformation has taken place should facilitate recognition. In natural circumstances, objects do not disappear and appear in different orientations inexplicably and an observer usually knows what to expect according to the context. This context includes the environment, and the history of the observers' movements, which specify the transient relationship between the object, the environment and the observer. We used interactive computer graphics to study the effects of providing observers with either implicit or explicit indications of their view transformations in the recognition of a class of shape found previously to be highly view-dependent. Results show that these cues aid recognition to varying degrees but mostly for oblique views and primarily in terms of accuracy not response times. These results provide evidence for egocentric encoding of shape and suggest that knowing ones' transformation in view helps to reduce the problem space involved in matching a shape percept with a mental representation.


Lack of awareness of partner STD risk among heterosexual couples.
Individuals' accurate assessment of their exposure to the risk of HIV and other STDs requires awareness of their sexual partners' risk behaviors and disease status. In a sample of 217 couples enrolled in a risk intervention trial in 1997-2002, both partners reported on their own risk behaviors and their perceptions of their partner's behavior; concordance of partners' reports was examined using kappa statistics. Individual and relationship characteristics predicting lack of awareness of a partner's risk behavior were explored using multivariate logistic regression. Three percent of women and 14% of men were unaware that their partner had recently had a concurrent partner. Eleven percent and 12%, respectively, were unaware that their partner had ever injected drugs; 10% and 12% were unaware that their partner had recently received an STD diagnosis; and 2% and 4% were unaware that their partner was HIV-positive. Women's lack of awareness of partner risk was associated with increasing age (odds ratio, 1.1), being of a race or ethnicity other than black or Latina (15.8) and having a Latino partner (3.7); it was positively associated with a man's report that he was married (4.4) and with relationship satisfaction as reported by both the woman and her partner (1.2 for each). Among men, lack of awareness was positively associated with partner's age (1.1) and with having a partner who was formerly married (8.2). Couple-based interventions that assess each partner's awareness of the other's risk behavior may help programs better target couples' STD prevention needs.


Benefits of Polydopamine as Particle/Matrix Interface in Polylactide/PD-BaSO4 Scaffolds.
This work reports the versatility of polydopamine (PD) when applied as a particle coating in a composite of polylactide (PLA). Polydopamine was observed to increase the particle-matrix interface strength and facilitate the adsorption of drugs to the material surface. Here, barium sulfate radiopaque particles were functionalized with polydopamine and integrated into a polylactide matrix, leading to the formulation of a biodegradable and X-ray opaque material with enhanced mechanical properties. Polydopamine functionalized barium sulfate particles also facilitated the adsorption and release of the antibiotic levofloxacin. Analysis of the antibacterial capacity of these composites and the metabolic activity and proliferation of human dermal fibroblasts in vitro demonstrated that these materials are non-cytotoxic and can be 3D printed to formulate complex biocompatible materials for bone fixation devices.


The effect of cathodic electrochemical potential of Ti-6Al-4V on cell viability: voltage threshold and time dependence.
The biocompatibility of metallic biomaterials is dependent on the redox state of the surface and its effect on cellular redox equilibrium. When metallic biomaterials experience mechanically assisted corrosion such as fretting, there is a drop in the voltage of its surface. Studies have demonstrated that cell viability is significantly degraded by sustained cathodic excursion in voltage of metallic biomaterials below a critical potential between -300 and -600 mV on commercially pure titanium. Cells cultured at above -300 mV showed little loss in viability whereas cells cultured on -600 mV Ti were almost 100% killed at 24 h. The goal of this study is to concisely define the voltage threshold and time-dependence of the cell killing effect seen on titanium surfaces. MC3T3 cells were cultured on electrochemically controlled Ti-6Al-4V surfaces at voltages ranging from -300 to -1000 mV for time periods ranging from 1 to 24 h. Cell viability and morphology was monitored with live-dead assay and scanning electron microscopy. Cell viability decreased from -300 to -400 mV and exhibited time-dependence where the more cathodic the potential, the faster the drop-off of viability. Hundred percent cell killing took as little as 4 h at -1000 mV and required 24 h at -400 mV. Sustained net cathodic currents with densities as low as 0.1 μA/cm(2) are observed during cell killing. This work shows reduction reactions are an important element of cellular viability in a time and potential dependent way and may explain why mechanically assisted corrosion reactions may lead to increased cell killing in metallic implants. Note: All voltages are versus Ag/AgCl.


Azole antifungals induce up-regulation of SAP4, SAP5 and SAP6 secreted proteinase genes in filamentous Candida albicans cells in vitro and in vivo.
Expression of fungal virulence factors can be influenced by exposure to antifungal agents. To test this hypothesis, we determined the effects of subinhibitory concentrations of three antifungal agents on expression of three secreted proteinase genes associated with virulence in filamentous forms of Candida albicans. GFP-SAP promoter constructs and fluorescence measurement, transcript profiling and RT-PCR in vitro and in an animal model of disseminated Candida infection. Exposure of C. albicans to subinhibitory concentrations of fluconazole in RPMI 1640 in the absence of serum led to up-regulation of the virulence-associated genes SAP4, SAP5 and SAP6 in hyphae and long pseudohyphae. Measurements with green fluorescent protein (GFP)-tagged promoters showed that the fluorescence of SAP4 and SAP6 under these conditions was strongest in the apical tip compartments of these filamentous cells and declined in compartments more proximal to the parent yeast cell. By contrast, SAP5-GFP fluorescence was expressed at similar levels in all cell compartments. Exposure to fluconazole led to significant increases in GFP-SAP4 and -SAP6 fluorescence in the filaments; itraconazole exposure also significantly increased GFP-SAP4 fluorescence, whereas flucytosine had no effect on any of the constructs. In experimentally infected animals, fluorescence of the GFP-SAP promoter fungal cells in kidney tissues was greater than that was seen in vitro for all four SAP constructs: treatment of animals with fluconazole did not significantly increase SAP promoter expression as measured by GFP fluorescence. Azole antifungal agents stimulated up-regulation of SAP4 and SAP6 genes in filamentous C. albicans cells in vitro and may therefore influence virulence as well as growth of the fungus. However, such effects appear to be transient in vivo.


Toll-like receptors, signaling adapters and regulation of the pro-inflammatory response by PI3K.
TLRs are a family of pattern recognition receptors that recognize conserved molecular structures/products from a wide variety of microbes. Following recognition of ligands, TLRs recruit signaling adapters to initiate a pro-inflammatory signaling cascade culminating in the activation of several transcription factor families. Additionally, TLR signals lead to activation of PI3K, affecting many aspects of the cellular response, including cell survival, proliferation and regulation of the pro-inflammatory response. The recent discovery of BCAP as a TLR signaling adaptor, crucial for linking TLRs to PI3K activation, allows new questions of the importance of PI3K activation downstream of TLRs. Here, we summarize the current understanding of signaling pathways activated by TLRs and provide our perspective on TLR mediated activation of PI3K and its impact on regulating cellular processes.


Structure-based interpretation of biotransformation pathways of amide-containing compounds in sludge-seeded bioreactors.
Partial microbial degradation of xenobiotic compounds in wastewater treatment plants (WWTPs) results in the formation of transformation products, which have been shown to be released and detectable in surface waters. Rule-based systems to predict the structures of microbial transformation products often fail to discriminate between alternate transformation pathways because structural influences on enzyme-catalyzed reactions in complex environmental systems are not well understood. The amide functional group is one such common substructure of xenobiotic compounds that may be transformed through alternate transformation pathways. The objective of this work was to generate a self-consistent set of biotransformation data for amide-containing compounds and to develop a metabolic logic that describes the preferred biotransformation pathways of these compounds as a function of structural and electronic descriptors. We generated transformation products of 30 amide-containing compounds in sludge-seeded bioreactors and identified them by means of HPLC-linear ion trap-orbitrap mass spectrometry. Observed biotransformation reactions included amide hydrolysis and N-dealkylation, hydroxylation, oxidation, ester hydrolysis, dehalogenation, nitro reduction, and glutathione conjugation. Structure-based interpretation of the results allowed for identification of preferences in biotransformation pathways of amides: primary amides hydrolyzed rapidly; secondary amides hydrolyzed at rates influenced by steric effects; tertiary amides were N-dealkylated unless specific structural moieties were present that supported other more readily enzyme-catalyzed reactions. The results allowed for the derivation of a metabolic logic that could be used to refine rule-based biotransformation pathway prediction systems to more specifically predict biotransformations of amide-containing compounds.