1,2-Dichlorohexafluoro-cyclobutane (1,2-c-C4F6Cl2, R-316c) a potent ozone depleting substance and greenhouse gas: atmospheric loss processes, lifetimes, and ozone depletion and global warming potentials for the (E) and (Z) stereoisomers.
The atmospheric processing of (E)- and (Z)-1,2-dichlorohexafluoro-cyclobutane (1,2-c-C4F6Cl2, R-316c) was examined in this work as the ozone depleting (ODP) and global warming (GWP) potentials of this proposed replacement compound are presently unknown. The predominant atmospheric loss processes and infrared absorption spectra of the R-316c isomers were measured to provide a basis to evaluate their atmospheric lifetimes and, thus, ODPs and GWPs. UV absorption spectra were measured between 184.95 to 230 nm at temperatures between 214 and 296 K and a parametrization for use in atmospheric modeling is presented. The Cl atom quantum yield in the 193 nm photolysis of R-316c was measured to be 1.90 ± 0.27. Hexafluorocyclobutene (c-C4F6) was determined to be a photolysis co-product with molar yields of 0.7 and 1.0 (±10%) for (E)- and (Z)-R-316c, respectively. The 296 K total rate coefficient for the O((1)D) + R-316c reaction, i.e., O((1)D) loss, was measured to be (1.56 ± 0.11) × 10(-10) cm(3) molecule(-1) s(-1) and the reactive rate coefficient, i.e., R-316c loss, was measured to be (1.36 ± 0.20) × 10(-10) cm(3) molecule(-1) s(-1) corresponding to a ~88% reactive yield. Rate coefficient upper-limits for the OH and O3 reaction with R-316c were determined to be <2.3 × 10(-17) and <2.0 × 10(-22) cm(3) molecule(-1) s(-1), respectively, at 296 K. The quoted uncertainty limits are 2σ and include estimated systematic errors. Local and global annually averaged lifetimes for the (E)- and (Z)-R-316c isomers were calculated using a 2-D atmospheric model to be 74.6 ± 3 and 114.1 ± 10 years, respectively, where the estimated uncertainties are due solely to the uncertainty in the UV absorption spectra. Stratospheric photolysis is the predominant atmospheric loss process for both isomers with the O((1)D) reaction making a minor, ~2% for the (E) isomer and 7% for the (Z) isomer, contribution to the total atmospheric loss. Ozone depletion potentials for (E)- and (Z)-R-316c were calculated using the 2-D model to be 0.46 and 0.54, respectively. Infrared absorption spectra for (E)- and (Z)-R-316c were measured at 296 K and used to estimate their radiative efficiencies (REs) and GWPs; 100-year time-horizon GWPs of 4160 and 5400 were obtained for (E)- and (Z)-R-316c, respectively. Both isomers of R-316c are shown in this work to be long-lived ozone depleting substances and potent greenhouse gases.


Brain abscesses: etiologic categorization with in vivo proton MR spectroscopy.
To compare the metabolite patterns observed at in vivo proton magnetic resonance (MR) spectroscopy of brain abscesses in patients for whom bacteriologic information was obtained from cultures and to categorize the MR spectral patterns with respect to the underlying etiologic agents. MR imaging and in vivo single-voxel proton MR spectroscopic data obtained from 75 patients with brain abscesses were retrospectively analyzed. Ex vivo spectroscopic experiments with the pus from 45 of these patients also were performed, and the data were further categorized on the basis of bacteriologic information. Quantification of various metabolites and metabolite ratios and statistical analyses of lactate and lactate/amino acid (AA) ratio levels were performed by using one-way analysis of variance. On the basis of in vivo proton MR spectroscopic and bacteriologic analysis findings, data were classified into three categories: Group 1 data showed resonances of lactate, AAs, and acetate, with or without succinate at proton MR spectroscopy; cultures for this group showed obligate anaerobes or a mixture of obligate and facultative anaerobes. The metabolite patterns in the group 2 and group 3 data were similar to the pattern of the group 1 data, with the exception that acetate and succinate resonances were absent. Culture was positive for either obligate aerobes or facultative anaerobes in group 2 and was sterile in group 3. At analysis of variance, in vivo data showed significant differences in lactate/AA ratios (P =.008), and ex vivo data showed significant differences in lactate levels (P =.001) among the three groups. It is possible to differentiate anaerobic from aerobic or sterile brain abscesses on the basis of metabolite patterns observed at in vivo proton MR spectroscopy. This information may be useful in facilitating prompt and appropriate treatment of patients with these abscesses.


#PrayForDad: Learning the Semantics Behind Why Social Media Users Disclose Health Information.
User-generated content in social media is increasingly acknowledged as a rich resource for research into health problems. One particular area of interest is in the semantics individuals evoke because they can influence when health-related information is disclosed. While there have been multiple investigations into why self-disclose occurs, much less is known about when individuals choose to disclose information about other people (e.g., a relative), which is a significant privacy concern. In this paper, we introduce a novel framework to investigate how semantics influence disclosure routines for 34 health issues. This framework begins with a supervised classification model to distinguish tweets that communicate personal health issues from confounding concepts (e.g., metaphorical statements that include a health-related keyword). Next, we annotate tweets for each health issue with linguistic and psychological categories (e.g. social processes, affective processes and personal concerns). Then, we apply a non-negative matrix factorization over a health issue-by-language category space. Finally, the factorized basis space is leveraged to group health issues into natural aggregations based around how they are discussed. We evaluate this framework with four months of tweets (over 200 million) and show that certain semantics correspond with whom a health mention pertains to. Our findings show that health issues related with family members, high medical cost and social support (e.g., Alzheimer's Disease, cancer, and Down syndrome) lead to tweets that are more likely to disclose another individual's health status, while tweets with more benign health issues (e.g., allergy, arthritis, and bronchitis) with biological processes (e.g., health and ingestion) and negative emotions are more likely to contain self-disclosures.


Natural killer cells act as rheostats modulating antiviral T cells.
Antiviral T cells are thought to regulate whether hepatitis C virus (HCV) and human immunodeficiency virus (HIV) infections result in viral control, asymptomatic persistence or severe disease, although the reasons for these different outcomes remain unclear. Recent genetic evidence, however, has indicated a correlation between certain natural killer (NK)-cell receptors and progression of both HIV and HCV infection, implying that NK cells have a role in these T-cell-associated diseases. Although direct NK-cell-mediated lysis of virus-infected cells may contribute to antiviral defence during some virus infections--especially murine cytomegalovirus (MCMV) infections in mice and perhaps HIV in humans--NK cells have also been suspected of having immunoregulatory functions. For instance, NK cells may indirectly regulate T-cell responses by lysing MCMV-infected antigen-presenting cells. In contrast to MCMV, lymphocytic choriomeningitis virus (LCMV) infection in mice seems to be resistant to any direct antiviral effects of NK cells. Here we examine the roles of NK cells in regulating T-cell-dependent viral persistence and immunopathology in mice infected with LCMV, an established model for HIV and HCV infections in humans. We describe a three-way interaction, whereby activated NK cells cytolytically eliminate activated CD4 T cells that affect CD8 T-cell function and exhaustion. At high virus doses, NK cells prevented fatal pathology while enabling T-cell exhaustion and viral persistence, but at medium doses NK cells paradoxically facilitated lethal T-cell-mediated pathology. Thus, NK cells can act as rheostats, regulating CD4 T-cell-mediated support for the antiviral CD8 T cells that control viral pathogenesis and persistence.


Open research practices: unintended consequences and suggestions for averting them. (Commentary on the Peer Reviewers' Openness Initiative).
The Peer Reviewers' Openness Initiative (PROI) is a move to enlist reviewers in the promotion of data-sharing. In this commentary, I discuss objections that can be raised, first to the specific proposals in the PROI, and second to data-sharing in general. I argue that although many objections have strong counter-arguments, others merit more serious consideration. Regarding the PROI, I suggest that it could backfire if editors and authors feel coerced into data-sharing and so may not be the most pragmatic way of encouraging greater openness. More generally, while promoting data-sharing, we need to be sensitive to cases where sharing of data from human participants could create ethical problems. Furthermore, those interested in promoting reproducible science need to defend against an increased risk of data-dredging when large, multivariable datasets are shared. I end with some suggestions to avoid these unintended consequences.


Medium-term temporal stability of the helminth component community structure in bank voles (Clethrionomys glareolus) from the Mazury Lake District region of Poland.
The structure of helminth communities in wild rodents is subject to seasonal variation, and is dependent on host age within years. Although between-year variation has been monitored, seldom has it been assessed rigorously by appropriate multifactorial analysis with potentially confounding factors taken into account. In this study we tested the null hypothesis that despite seasonal, host age and sex effects, helminth communities should show relative stability between years. Over a period of 3 years (1998-2000) we sampled bank vole (Clethrionomys glareolus) populations (total n = 250) at 2 points in the year: in spring, at the start of the breeding season, and in autumn, after the cessation of breeding. In spite of seasonal differences and strong age effects, the between-year effects were surprisingly small. Measures of component community structure (Berger-Parker dominance index, the dominant species, S. petrusewiczi) did not vary, or varied only slightly from year to year. The majority of measures of infracommunity structure [Brillouin's index of diversity, prevalence of all helminths combined, prevalence and abundance of H. mixtum (the most prevalent helminth), mean species richness] did not differ significantly between years when other factors such as age, sex and seasonal variation had been taken into account. Some between-year variations were found (at the component community level, Simpson's index of diversity; at the infracommunity level, prevalence and abundance of S. petrusewiczi and abundance of all helminths combined), but even these were modest in comparison to seasonal and age differences, and were primarily attributable to S. petrusewiczi. We conclude that despite dynamic within-year fluctuations, helminth communities in bank voles in this region of Poland show relative stability across years. The sporadic occurrence of individual platyhelminths at low prevalence, makes little difference to the overall structure, which is largely maintained by the key roles played by the dominant intestinal nematodes of bank voles and the rarer species collectively.


Spontaneous Rayleigh-Brillouin scattering of ultraviolet light in nitrogen, dry air, and moist air.
Atmospheric lidar techniques for the measurement of wind, temperature, and optical properties of aerosols rely on the exact knowledge of the spectral line shape of the scattered laser light on molecules. We report on spontaneous Rayleigh-Brillouin scattering measurements in the ultraviolet at a scattering angle of 90 degrees on N(2) and on dry and moist air. The measured line shapes are compared to the Tenti S6 model, which is shown to describe the scattering line shapes in air at atmospheric pressures with small but significant deviations. We demonstrate that the line profiles of N(2) and air under equal pressure and temperature conditions differ significantly, and that this difference can be described by the S6 model. Moreover, we show that even a high water vapor content in air up to a volume fraction of 3.6vol.% has no influence on the line shape of the scattered light. The results are of relevance for the future spaceborne lidars on ADM-Aeolus (Atmospheric Dynamics Mission) and EarthCARE (Earth Clouds, Aerosols, and Radiation Explorer).


Connectivity-based parcellation of human cingulate cortex and its relation to functional specialization.
Whole-brain neuroimaging studies have demonstrated regional variations in function within human cingulate cortex. At the same time, regional variations in cingulate anatomical connections have been found in animal models. It has, however, been difficult to estimate the relationship between connectivity and function throughout the whole cingulate cortex within the human brain. In this study, magnetic resonance diffusion tractography was used to investigate cingulate probabilistic connectivity in the human brain with two approaches. First, an algorithm was used to search for regional variations in the probabilistic connectivity profiles of all cingulate cortex voxels with the whole of the rest of the brain. Nine subregions with distinctive connectivity profiles were identified. It was possible to characterize several distinct areas in the dorsal cingulate sulcal region. Several distinct regions were also found in subgenual and perigenual cortex. Second, the probabilities of connection between cingulate cortex and 11 predefined target regions of interest were calculated. Cingulate voxels with a high probability of connection with the different targets formed separate clusters within cingulate cortex. Distinct connectivity fingerprints characterized the likelihood of connections between the extracingulate target regions and the nine cingulate subregions. Last, a meta-analysis of 171 functional studies reporting cingulate activation was performed. Seven different cognitive conditions were selected and peak activation coordinates were plotted to create maps of functional localization within the cingulate cortex. Regional functional specialization was found to be related to regional differences in probabilistic anatomical connectivity.


miR-511-3p modulates genetic programs of tumor-associated macrophages.
Expression of the mannose receptor (MRC1/CD206) identifies macrophage subtypes, such as alternatively activated macrophages (AAMs) and M2-polarized tumor-associated macrophages (TAMs), which are endowed with tissue-remodeling, proangiogenic, and protumoral activity. However, the significance of MRC1 expression for TAM's protumoral activity is unclear. Here, we describe and characterize miR-511-3p, an intronic microRNA (miRNA) encoded by both mouse and human MRC1 genes. By using sensitive miRNA reporter vectors, we demonstrate robust expression and bioactivity of miR-511-3p in MRC1(+) AAMs and TAMs. Unexpectedly, enforced expression of miR-511-3p tuned down the protumoral gene signature of MRC1(+) TAMs and inhibited tumor growth. Our findings suggest that transcriptional activation of Mrc1 in TAMs evokes a genetic program orchestrated by miR-511-3p, which limits rather than enhances their protumoral functions. Besides uncovering a role for MRC1 as gatekeeper of TAM's protumoral genetic programs, these observations suggest that endogenous miRNAs may operate to establish thresholds for inflammatory cell activation in tumors.


Association between county level cannabis dispensary counts and opioid related mortality rates in the United States: panel data study.
To examine county level associations between the prevalence of medical and recreational cannabis stores (referred to as dispensaries) and opioid related mortality rates. Panel regression methods. 812 counties in the United States in the 23 states that allowed legal forms of cannabis dispensaries to operate by the end of 2017. The study used US mortality data from the Centers for Disease Control and Prevention combined with US census data and data from Weedmaps.com on storefront dispensary operations. Data were analyzed at the county level by using panel regression methods. The main outcome measures were the log transformed, age adjusted mortality rates associated with all opioid types combined, and with subcategories of prescription opioids, heroin, and synthetic opioids other than methadone. The associations of medical dispensary and recreational dispensary counts with age adjusted mortality rates were also analyzed. County level dispensary count (natural logarithm) is negatively related to the log transformed, age adjusted mortality rate associated with all opioid types (β=-0.17, 95% confidence interval -0.23 to -0.11). According to this estimate, an increase from one to two storefront dispensaries in a county is associated with an estimated 17% reduction in all opioid related mortality rates. Dispensary count has a particularly strong negative association with deaths caused by synthetic opioids other than methadone (β=-0.21, 95% confidence interval -0.27 to -0.14), with an estimated 21% reduction in mortality rates associated with an increase from one to two dispensaries. Similar associations were found for medical versus recreational storefront dispensary counts on synthetic (non-methadone) opioid related mortality rates. Higher medical and recreational storefront dispensary counts are associated with reduced opioid related death rates, particularly deaths associated with synthetic opioids such as fentanyl. While the associations documented cannot be assumed to be causal, they suggest a potential association between increased prevalence of medical and recreational cannabis dispensaries and reduced opioid related mortality rates. This study highlights the importance of considering the complex supply side of related drug markets and how this shapes opioid use and misuse.


Access to palliative care by disease trajectory: a population-based cohort of Ontario decedents.
To examine access to palliative care between different disease trajectories and compare to other geographic areas. A retrospective population-based decedent cohort study using linked administrative data. Ontario, Canada. Ontario decedents between 1 April 2010 and 31 December 2012. Patients were categorised into disease trajectories: terminal illness (eg, cancer), organ failure (eg, chronic heart failure), frailty (eg, dementia), sudden death or other. Receipt of palliative care services from institutional and community settings, derived from a validated list of palliative care codes from multiple administrate databases. Receiving any palliative care services in the last year of life (yes/no), intensity (total days) and time of initiation of palliative care, in hospital and community sectors. Multivariable analysis examined the association between disease trajectory and the receipt of palliative care in the last year of life. We identified 235 159 decedents in Ontario. In the last year of life, 88% of terminal illness, 44% of organ failure and 32% of frailty decedents accessed at least one palliative care service. Most care was provided during an inpatient hospitalisation. Terminal illness decedents received twice as many palliative care days (mean of 49 days) compared with organ failure and frailty decedents. Patients with terminal illness initiated palliative care median of 107 days before death compared with median of 19 days among those using the US Medicare hospice benefit. Terminal illness decedents are more likely to receive any palliative care, with increased intensity and earlier before death than organ failure or frailty decedents. These data serve as a useful comparison for other countries with similar and different healthcare systems and eligibility criteria.


Systematically describing gross lesions in corals.
Many coral diseases are characterized based on gross descriptions and, given the lack or difficulty of applying existing laboratory tools to understanding causes of coral diseases, most new diseases will continued to be described based on appearance in the field. Unfortunately, many existing descriptions of coral disease are ambiguous or open to subjective interpretation, making comparisons between oceans problematic. One reason for this is that the process of describing lesions is often confused with that of assigning causality for the lesion. However, causality is usually something not obtained in the field and requires additional laboratory tests. Because a concise and objective morphologic description provides the foundation for a case definition of any disease, there is a need for a consistent and standardized process to describe lesions of corals that focuses on morphology. We provide a framework to systematically describe and name diseases in corals involving 4 steps: (1) naming the disease, (2) describing the lesion, (3) formulating a morphologic diagnosis and (4) formulating an etiologic diagnosis. This process focuses field investigators on describing what they see and separates the process of describing a lesion from that of inferring causality, the latter being more appropriately done using laboratory techniques.


Direct detection of ultralow trace amounts of isocyanates in air using a fluorescent conjugated polymer.
A fluorescence sensory polymer containing the pentiptycene and tetraphenylethylene units linked by acetylene was synthesized for direct detection of isocyanates in air. Eight industrially available aliphatic and aromatic isocyanates were tested. The polymer film shows a rapid fluorescence quenching response to any type of isocyanates with a ppt level of detection limit, which is much lower than the permissible exposure limit of 5 ppb.


Consumption of hydrogen-rich water alleviates renal injury in spontaneous hypertensive rats.
In hypertensive animals and patients, oxidative stress represents the primary risk factor for progression of renal disease. Recently, it has been demonstrated that hydrogen, as a novel antioxidant, can selectively reduce hydroxyl radicals and peroxynitrite anion to exert therapeutic antioxidant activity. Herein, we investigated the protective effect of hydrogen-rich water (HW) against renal injury in spontaneously hypertensive rats (SHR). The 8-week-old male SHR and age-matched Wistar-Kyoto rats were randomized into HW-treated (1.3 ± 0.2 mg/l for 3 months, drinking) and vehicle-treated group. Although treatment with HW had no significant effect on blood pressure, it significantly ameliorated renal injury in SHR. Treatment with HW lowered reactive oxygen species formation, upregulated the activities of superoxide dismutase, glutathione peroxidase, glutathione-S-epoxide transferase, and catalase, and suppressed NADPH oxidase activity. Treatment with HW in SHR depressed pro-inflammatory cytokines expression including TNF-α, IL-6, IL-1β, and macrophage chemoattractant protein 1, which might be mediated by suppressing nuclear factor-κB activation. In addition, treatment with HW had protective effect on mitochondrial function including adenosine triphosphate formation and membrane integrity in SHR. In conclusion, consumption of HW is a promising strategy to alleviate renal injury as a supplement for anti-hypertensive therapy.


First Report of Pine Wilt Disease Caused by Bursaphelenchus xylophilus on Pinus thunbergii in the Inland City of Zibo, Shandong, China.
The pinewood nematode (PWN) Bursaphelenchus xylophilus (Steiner & Buhrer 1934) Nickle 1970 is the causal agent of pine wilt disease. It is especially damaging in East Asian countries, including Japan, China, and Korea. In China, the nematode has been found in Anhui, Guangdoung, Guizhou, Chongqing, and Zhejiang Provinces since its discovery in Jiangsu Province in 1982 (1). China is confronted with an enormous threat to its pine forests. B. xylophilus is transmitted by the insect vector pine sawyer beetle (Monochamus alternatus). The main host trees are Pinus massoniana, P. thunbergii, and P. densiflora, which are the most common pine trees in China. Shandong Province, located north of Jiangsu Province, is a high-risk area because it was thought to be the northernmost suitable area for the pine wood nematode. P. tabulaeformis, P. densiflora, and P. thunbergii are the principal hosts. In 2010, a pine tree with suspected wilt disease was found in Lushang Forest (36°16'31.11″ N, 118°03'59.79″ E) of P. thunbergii located in Zibo city of Shandong Province. Symptoms were systemic, with almost all leaves brown or yellowish; the tree was nearly dead. Wood samples were collected and nematodes were extracted using a modified Baermann's funnel method. After 12 h, the nematodes were collected from the wood chips, and their morphology was observed with an inverted light microscope (Nikon 90i, Japan). Nematodes had a typical Aphelenchoid-type esophagus and female vulva flap. Females had subcylindrical tails, usually with broadly rounded terminus, some with a short mucro, and flat vulva, whereas males had large paired arcuate spicules with a sharply pointed prominent rostrum, and typical disc-like expansions on distal ends. Standard measurements of these nematodes were as follows: 25 females: body length = 960.9 ± 117.4 (791.5 to 1,265.2) μm, a = 32.1 ± 5.1 (23.7 to 44.5), b = 13.6 ± 1.4 (11.4 to 16.1), c = 28.3 ± 4.6 (21.7 to 42.2), V = 77.8 ± 2.0 (74.2 to 83.9), stylet length = 13.7 ± 1.6 (11.4 to 17.6) μm; 21 males: body length = 785.6 ± 103.2 (609.6 to 1,004.5) μm, a = 33.3 ± 4.4 (26.0 to 40.8), b = 11.9 ± 1.3 (9.0 to 14.6), c = 31.0 ± 2.7 (25.5 to 37.1), stylet length = 13.5 ± 1.9 (11.0 to 17.5) μm, spicule length = 18.8 ± 2.5 (14.9 to 23.9) μm. The morphometrics of this population, apart from body length and "a" value, which are shorter than the Portugal isolate measured by Mota et al. (3), are very much in the same range reported for B. xylophilus. For a more accurate identification, DNA was extracted from individual nematodes using a liquid nitrogen method. The internal transcribed spacers (ITS-1, ITS-2, 5.8S) were amplified by using PCR (2). Nucleotide sequences were compared with the sequences of B. xylophilus in GenBank, accession nos. JN684828 (Portugal), JN684829 (Portugal), JF826219 (Madeira Island) and JQ288086 (Japan). The ITS DNA sequences of the nematode from P. thunbergii were 99% identical to those of B. xylophilus in GenBank. A sequence of this nematode was submitted to the GenBank database and assigned the number KC460340. We have thus confirmed that B. xylophilus is now present north of Changjiang River in Zibo city, Shandong Province. This range expansion, perhaps the result of global warming, will affect both domestic and international quarantine efforts to control the further spread of pinewood nematode. References: (1) X. Y. Cheng et al. Heredity 100:356, 2008. (2) K. Metge and W. Burgermeister. J. Plant Dis. Protect. 113:275, 2006. (3) M. Mota et al. Nematology 1:727, 1999.


Ganglioside composition of differentiated Caco-2 cells resembles human colostrum and neonatal rat intestine.
Gangliosides are glycosphingolipids found in cell membranes and human milk with important roles in cell proliferation, differentiation, growth, adhesion, migration, signalling and apoptosis. Similar changes in ganglioside composition occur during embryonic development, lactation and cancer cell differentiation. It is not known, however, whether ganglioside compositional changes that occur in differentiating colon cancer cells reflect changes that occur during intestinal development. The Caco-2 cell line is commonly used to study physiological and pathophysiological processes in the small intestine and colon. Therefore, to examine this question, undifferentiated and differentiated Caco-2 cells were grown and total lipid was extracted from cell supernatant fractions using the Folch method. The upper aqueous phase containing gangliosides was collected and purified. Total gangliosides were measured as ganglioside-bound N-acetyl neuraminic acid, while individual ganglioside content was quantified via a colorimetric assay for sialic acid and scanning densitometry. The total ganglioside content of differentiated Caco-2 cells was 2.5 times higher compared with undifferentiated cells. Differentiated Caco-2 cells had significantly more (N-acetylneuraminyl) 2-galactosylglucosyl ceramide (GD3) and polar gangliosides, and a lower N-acetylneuraminylgalactosylglucosylceramide (GM3):GD3 ratio than undifferentiated cells. The present study demonstrates that the total ganglioside content and individual ganglioside composition of differentiated Caco-2 cells are similar to those of human colostrum and neonatal rat intestine. Differentiated Caco-2 cells may therefore be an alternative model for studying physiological and pathological processes in the small intestine and colon, and may help to elucidate possible functions for specific gangliosides in development and differentiation. Further research using more sensitive techniques of ganglioside analysis is needed to confirm these findings.


Investigations on the elasticity of functional gold nanoparticles using single-molecule force spectroscopy.
A wide range of investigation tools and frameworks aimed at the in depth understanding of the physico-chemical properties of different nanomaterials and at exploring their cellular interactions and effects have been reported in the past couple of decades. Among these, Single-Molecule Force Spectroscopy (SMFS) emerges as a very important tool for characterizing nanoparticles (NPs) and one of its very valuable applications consists in the quantitative analysis of the NPs' elasticity. In SMFS experiments that tackle this subject, a sharp tip present on the apex of a cantilever is indented into a single NP, and then the Young's modulus is determined as a measure of its elasticity, which is one of the fundamental mechanical parameters affecting the structural and functional cellular parameters. Based on such approaches, SMFS enables the observation and analysis of significant cellular effects that are relevant to various cellular parameters. In this focused review, we turn our attention towards several approaches for detecting the elasticity of NPs, systematically summarizing the divergent elasticity values of distinct gold nanoparticles (AuNPs) with different surfaces. We carry as well a critical discussion on the elasticity assessment models and the fundamental factors that influence NP elasticity assessment by means of SMFS.


Mutation of HES7 in a large extended family with spondylocostal dysostosis and dextrocardia with situs inversus.
Spondylocostal dysotosis (SCD) is a rare developmental congenital abnormality of the axial skeleton. Mutation of genes in the Notch signaling pathway cause SCD types 1-5. Dextrocardia with situs inversus is a rare congenital malformation in which the thoracic and abdominal organs are mirror images of normal. Such laterality defects are associated with gene mutations in the Nodal signaling pathway or cilia assembly or function. We investigated two distantly related individuals with a rare combination of severe segmental defects of the vertebrae (SDV) and dextrocardia with situs inversus. We found that both individuals were homozygous for the same mutation in HES7, and that this mutation caused a significant reduction of HES7 protein function; HES7 mutation causes SCD4. Two other individuals with SDV from two unrelated families were found to be homozygous for the same mutation. Interestingly, although the penetrance of the vertebral defects was complete, only 3/7 had dextrocardia with situs inversus, suggesting randomization of left-right patterning. Two of the affected individuals presented with neural tube malformations including myelomeningocele, spina bifida occulta and/or Chiari II malformation. Such neural tube phenotypes are shared with the originally identified SCD4 patient, but have not been reported in the other forms of SCD. In conclusion, it appears that mutation of HES7 is uniquely associated with defects in vertebral, heart and neural tube formation, and this observation will help provide a discriminatory diagnostic guide in patients with SCD, as well as inform molecular genetic testing.


Evaluation of contractile phenotype in airway smooth muscle cells isolated from endobronchial biopsy and tissue specimens from horses.
OBJECTIVE To develop a method to maintain the initial phenotype of airway smooth muscle (ASM) cells isolated from equine endobronchial biopsy specimens in long-term cell culture. SAMPLE Endobronchial tissue specimens (8 to 10/horse) collected from the lungs of previously healthy horses at necropsy (n = 12) and endobronchial biopsy specimens collected from standing, sedated, heaves-affected horses in clinical remission of the disease (5) and control horses (4). PROCEDURES A sampling protocol was developed to recover and maintain a contractile phenotype in ASM cells from endobronchial specimens from freshly harvested equine lungs and from healthy and heaves-affected horses. Immunologic techniques were used to evaluate the contractile phenotype of ASM cells in culture. RESULTS Characteristic ASM cells were successfully cultured from endobronchial tissue or biopsy specimens from both healthy and heaves-affected horses, and their contractile phenotype was maintained for up to 7 passages. Moreover, the capacity of cells at the seventh passage to contract in a collagen gel in response to methacholine was maintained. CONCLUSIONS AND CLINICAL RELEVANCE ASM cells isolated from equine endobronchial tissue and biopsy specimens were able to maintain a contractile phenotype in long-term cell cultures, suggesting they could be used for tissue engineering and in vitro studies of equine ASM cells.


A molecular barcode to inform the geographical origin and transmission dynamics of Plasmodium vivax malaria.
Although Plasmodium vivax parasites are the predominant cause of malaria outside of sub-Saharan Africa, they not always prioritised by elimination programmes. P. vivax is resilient and poses challenges through its ability to re-emerge from dormancy in the human liver. With observed growing drug-resistance and the increasing reports of life-threatening infections, new tools to inform elimination efforts are needed. In order to halt transmission, we need to better understand the dynamics of transmission, the movement of parasites, and the reservoirs of infection in order to design targeted interventions. The use of molecular genetics and epidemiology for tracking and studying malaria parasite populations has been applied successfully in P. falciparum species and here we sought to develop a molecular genetic tool for P. vivax. By assembling the largest set of P. vivax whole genome sequences (n = 433) spanning 17 countries, and applying a machine learning approach, we created a 71 SNP barcode with high predictive ability to identify geographic origin (91.4%). Further, due to the inclusion of markers for within population variability, the barcode may also distinguish local transmission networks. By using P. vivax data from a low-transmission setting in Malaysia, we demonstrate the potential ability to infer outbreak events. By characterising the barcoding SNP genotypes in P. vivax DNA sourced from UK travellers (n = 132) to ten malaria endemic countries predominantly not used in the barcode construction, we correctly predicted the geographic region of infection origin. Overall, the 71 SNP barcode outperforms previously published genotyping methods and when rolled-out within new portable platforms, is likely to be an invaluable tool for informing targeted interventions towards elimination of this resilient human malaria.


Combined chemotherapy and radiation therapy for cervical cancer.
More than 50% of patients with newly diagnosed cervical cancer will undergo radiation therapy as a primary treatment modality. Results from five phase III trials conducted by National Cancer Institute (NCI)-sponsored cooperative groups showed significant survival advantages from the addition of cisplatin-based chemotherapy to primary radiation therapy, changing the standard of care for patients with bulky or locally advanced cervical carcinoma. The majority of patients for whom treatment fails, however, have persistent pelvic disease. Other agents (alone or in combination with cisplatin) with potential synergy with radiation therapy should be studied. Future trials should investigate potential interrelationships between anemia, tumor hypoxia, angiogenesis, and the effectiveness of radiation therapy. Therapies specific to tumor hypoxia (tirapazamine) or strategies to maintain higher nadir hemoglobin levels during treatment (aggressive transfusion policies; erythropoietin) should also be studied.


Signaling through dynamic linkers as revealed by PKA.
Protein kinase A (PKA) is a prototype of multidomain signaling proteins functioning as allosteric conformational switches. Allosteric transitions have been the subject of extensive structural and dynamic investigations focusing mainly on folded domains. However, the current understanding of the allosteric role of partially unstructured linkers flanking globular domains is limited. Here, we show that a dynamic linker in the regulatory subunit (R) of PKA serves not only as a passive covalent thread, but also as an active allosteric element that controls activation of the kinase subunit (C) by tuning the inhibitory preequilibrium of a minimally populated intermediate (apo R). Apo R samples both C-binding competent (inactive) and incompetent (active) conformations within a nearly degenerate free-energy landscape and such degeneracy maximally amplifies the response to weak (∼2RT), but conformation-selective interactions elicited by the linker. Specifically, the R linker that in the R:C complex docks in the active site of C in apo R preferentially interacts with the C-binding incompetent state of the adjacent cAMP-binding domain (CBD). These unanticipated findings imply that the formation of the intermolecular R:C inhibitory interface occurs at the expense of destabilizing the intramolecular linker/CBD interactions in R. A direct implication of this model, which was not predictable solely based on protein structure, is that the disruption of a linker/CBD salt bridge in the R:C complex unexpectedly leads to increased affinity of R for C. The linker includes therefore sites of R:C complex frustration and frustration-relieving mutations enhance the kinase inhibitory potency of R without compromising its specificity.


The impact of computer science in molecular medicine: enabling high-throughput research.
The Human Genome Project and the explosion of high-throughput data have transformed the areas of molecular and personalized medicine, which are producing a wide range of studies and experimental results and providing new insights for developing medical applications. Research in many interdisciplinary fields is resulting in data repositories and computational tools that support a wide diversity of tasks: genome sequencing, genome-wide association studies, analysis of genotype-phenotype interactions, drug toxicity and side effects assessment, prediction of protein interactions and diseases, development of computational models, biomarker discovery, and many others. The authors of the present paper have developed several inventories covering tools, initiatives and studies in different computational fields related to molecular medicine: medical informatics, bioinformatics, clinical informatics and nanoinformatics. With these inventories, created by mining the scientific literature, we have carried out several reviews of these fields, providing researchers with a useful framework to locate, discover, search and integrate resources. In this paper we present an analysis of the state-of-the-art as it relates to computational resources for molecular medicine, based on results compiled in our inventories, as well as results extracted from a systematic review of the literature and other scientific media. The present review is based on the impact of their related publications and the available data and software resources for molecular medicine. It aims to provide information that can be useful to support ongoing research and work to improve diagnostics and therapeutics based on molecular-level insights.


Dearomative [2,3] sigmatropic rearrangement of ammonium ylides followed by 1,4-elimination to form α-(ortho-vinylphenyl)amino acid esters.
A base-induced dearomative [2,3] sigmatropic rearrangement of amino acid ester-derived ammonium salts followed by 1,4-elimination produced α-(ortho-vinylphenyl)amino acid esters. The reaction of azetidine-2-carboxylic acid-derived ammonium salt, (1S,2S,1'R)-3b, proceeded with a perfect N-to-C chirality transfer to afford α-(ortho-vinylphenyl)azetidine-2-carboxylic acid ester, (R)-5 (99% ee). On the other hand, the reaction of glycine-derived ammonium salt (R)-6a, which involves an efficient chirality transfer from a chiral benzylic carbon to an α-carbon of an ester carbonyl giving the optically active α-(ortho-vinylphenyl)glycine ester, (R)-8a (85% ee), was demonstrated. Although this dearomative [2,3] rearrangement followed by 1,4-elimination has limitations with regard to the structures of the substrates, our method provides unique access to substituted α-arylamino acid derivatives.


The pseudorabies virus VP22 homologue (UL49) is dispensable for virus growth in vitro and has no effect on virulence and neuronal spread in rodents.
The tegument of herpesvirus virions is a distinctive structure whose assembly and function are not well understood. The herpes simplex virus type 1 VP22 tegument protein encoded by the UL49 gene is conserved among the alphaherpesviruses. Using cell biology and viral genetics, we provide an initial characterization of the pseudorabies virus (PRV) VP22 homologue. We identified three isoforms of VP22 present in PRV-infected cells that can be resolved by polyacrylamide gel electrophoresis. The predominant form is not phosphorylated and is present in virions, while the other two species are phosphorylated and excluded from virions. VP22 localized to the nucleus by 6 h postinfection, as determined by immunofluorescence and cell fractionation. VP22 immunofluorescence in the nucleus was both diffuse and in punctate structures. The punctate nuclear localization was the most pronounced form of staining and did not localize exclusively to sites of viral DNA replication. Unexpectedly, a VP22 null mutant had no obvious phenotypes during tissue culture infections and was similar to the wild type in all respects. Moreover, the VP22 null mutant was as virulent and neuroinvasive as the wild-type virus after infection of the rodent eye and spread to the brain using both anterograde and retrograde neuronal circuits.


Phagosomal rupture by Mycobacterium tuberculosis results in toxicity and host cell death.
Survival within macrophages is a central feature of Mycobacterium tuberculosis pathogenesis. Despite significant advances in identifying new immunological parameters associated with mycobacterial disease, some basic questions on the intracellular fate of the causative agent of human tuberculosis in antigen-presenting cells are still under debate. To get novel insights into this matter, we used a single-cell fluorescence resonance energy transfer (FRET)-based method to investigate the potential cytosolic access of M. tuberculosis and the resulting cellular consequences in an unbiased, quantitative way. Analysis of thousands of THP-1 macrophages infected with selected wild-type or mutant strains of the M. tuberculosis complex unambiguously showed that M. tuberculosis induced a change in the FRET signal after 3 to 4 days of infection, indicating phagolysosomal rupture and cytosolic access. These effects were not seen for the strains M. tuberculosisΔRD1 or BCG, both lacking the ESX-1 secreted protein ESAT-6, which reportedly shows membrane-lysing properties. Complementation of these strains with the ESX-1 secretion system of M. tuberculosis restored the ability to cause phagolysosomal rupture. In addition, control experiments with the fish pathogen Mycobacterium marinum showed phagolysosomal translocation only for ESX-1 intact strains, further validating our experimental approach. Most importantly, for M. tuberculosis as well as for M. marinum we observed that phagolysosomal rupture was followed by necrotic cell death of the infected macrophages, whereas ESX-1 deletion- or truncation-mutants that remained enclosed within phagolysosomal compartments did not induce such cytotoxicity. Hence, we provide a novel mechanism how ESX-1 competent, virulent M. tuberculosis and M. marinum strains induce host cell death and thereby escape innate host defenses and favor their spread to new cells. In this respect, our results also open new research directions in relation with the extracellular localization of M. tuberculosis inside necrotic lesions that can now be tackled from a completely new perspective.


Management problems in acute hydrocephalus after subarachnoid hemorrhage.
In a consecutive series of 473 patients admitted within 72 hours after a subarachnoid hemorrhage, 91 (19%) had hydrocephalus on the initial computed tomogram. Consciousness was unimpaired in 25 of the 91 (28%). In 11 more patients acute hydrocephalus developed within 1 week after subarachnoid hemorrhage. Thirty-eight (8%) of all 473 patients subsequently showed clinical deterioration because of acute hydrocephalus; 11 of these 38 had fluctuations in the level of consciousness. Of the 66 patients with acute hydrocephalus and impaired consciousness on admission, 26 (39%) spontaneously improved within 24 hours. Ventricular drainage was performed in 32 (31%) of the 102 patients with acute hydrocephalus (7% of all 473 patients). Consciousness improved after ventricular drainage in 25 (78%) of the 32 patients. Ventriculitis developed in 12 of the 24 patients with external drainage, mainly after greater than 3 days of drainage, and in none of the eight patients with an internal shunt. Among the 340 patients with aneurysmal subarachnoid hemorrhage and no long-term tranexamic acid treatment, the frequency of rebleeding in patients with ventricular drainage (43% of 23) was significantly higher than in hydrocephalic patients without drainage (15% of 52 patients; chi 2 = 5.009, p = 0.025) and patients without acute hydrocephalus (20% of 265 patients; chi 2 = 5.521, p = 0.019). We conclude that spontaneous improvement occurs in half of the patients with acute hydrocephalus and impaired consciousness on admission, which is usually apparent within 24 hours, and that the outcome of patients who need ventricular drainage will improve if rebleeding and infection after insertion of the ventricular drain can be prevented.


Age-Dependent Changes in Transcription Factor FOXO Targeting in Female Drosophila.
FOXO transcription factors have long been associated with longevity control and tissue homeostasis. Although the transcriptional regulation of FOXO have been previously characterized (especially in long-lived insulin mutants and under stress conditions), how normal aging impacts the transcriptional activity of FOXO is poorly understood. Here, we conducted a chromatin immunoprecipitation sequencing (ChIP-Seq) analysis in both young (2-week-old) and aged (5-week-old) wild-type female fruit flies, Drosophila melanogaster, to evaluate the dynamics of FOXO gene targeting during aging. Intriguingly, the number of FOXO-bound genes dramatically decreases with age (from 2617 to 224). Consistent to the reduction of FOXO binding activity, many genes targeted by FOXO in young flies are transcriptionally altered with age, either up-regulated (FOXO-repressing genes) or down-regulated (FOXO-activating genes) in adult head tissue. In addition, we show that many FOXO-bound genes in wild-type flies are unique from those in insulin receptor substrate chico mutants. Distinct from chico mutants, FOXO targets specific cellular processes (e.g., actin cytoskeleton) and signaling pathways (e.g., Hippo, MAPK) in young wild-type female flies. FOXO targeting on these pathways decreases with age. Interestingly, FOXO targets in aged flies are enriched in cellular processes like chromatin organization and nucleosome assembly. Furthermore, FOXO binding to core histone genes is well maintained at aged flies. Together, our findings provide new insights into dynamic FOXO targeting under normal aging and highlight the diverse and understudied regulatory mechanisms for FOXO transcriptional activity.


CD82 expression alters with human endometrial cycles and affects the uterine endometrial receptivity in vitro.
Embryo implantation is a process that requires both temporal and spatial synchronization of the uterine endometrium and the embryo, and the endometrium becomes receptive to the embryo during the window of implantation. Although the expression patterns of many implantation-related molecules change dynamically during this process, the impact of CD82 on endometrial receptivity has not been elucidated. By immunohistochemical staining, we found that CD82 levels rose from the proliferative phase to the secretory phase in human endometrium. Specifically, the highest level appeared in mid- and late-secretory phases. Consistently, RL95-2 cells, representative of high-receptive endometrial epithelium, expressed higher levels of CD82 than did HEC-1A cells, which are representative of low-receptive endometrial epithelium, as detected by reverse transcription-polymerase chain reaction, Western blot and immunofluorescence. Furthermore, progesterone up-regulated the expression of CD82 in both epithelial cell lines. Down-regulation of CD82 in RL95-2 cells by either CD82 siRNA transfection or treatment with a CD82 antibody significantly decreased the adhesion of human embryonic JAR cells to RL95-2 cell monolayers (P < 0.01) and inhibited the phosphorylation of focal adhesion kinase (FAK). In contrast, up-regulation of CD82 in HEC-1A cells by CD82 cDNA transfection promoted embryonic JAR cell adhesion to HEC-1A monolayers (P < 0.05) and activated the phosphorylation of FAK. In conclusion, the expression of CD82 increases in endometrial tissues during the window of embryo implantation, CD82 expression affects endometrial receptivity of the uterine epithelial cells in vitro, and the FAK signaling pathway may be involved in this phenomenon. The correlation between CD82 and endometrial receptivity suggests that CD82 may serve as a potential marker of endometrial function.


Cell encapsulation and oxygenation in nanoporous microcontainers.
With strides in stem cell biology, cell engineering and molecular therapy, the transplantation of cells to produce therapeutic molecules endogenously is an attractive and achievable alternative to the use of exogenous drugs. The encapsulation of such cell transplants in semi-permeable, nanoporous constructs is often required to protect them from immune attack and to prevent their proliferation in the host. However, effective graft immunoisolation has been mostly elusive owing to the absence of a high-throughput method to create precisely controlled, high-aspect-ratio nanopores. To address the clinical need for effective cell encapsulation and immunoisolation, we devised a biocompatible cell-encapsulating microcontainer and a method to create highly anisotropic nanopores in the microcontainer's surface. To evaluate the efficacy of these nanopores in oxygenating the encapsulated cells, we engineered 9L rat glioma cells to bioluminesce under hypoxic conditions. The methods described above should aid in evaluating the long term survival and efficacy of cellular grafts.


An anatomical and clinical review of cricopharyngeal achalasia in the dog.
Cricopharyngeal achalasia is a rare cause of dysphagia in the dog. However it must be differentiated from other causes of dysphagia as it is treatable with surgery. It is a disruption of the cricopharyngeal phase of the oropharyngeal phase of deglutition. There appears to be an incoordination in the swallowing process between the relaxation of the rostral, middle pharyngeal muscles and the caudal pharyngeal muscles. It is seen as a primary condition in young animals presenting soon after weaning onto solid food. The dogs appear clinically healthy unless there is secondary aspiration pneumonia or emaciation. These dogs may present as respiratory emergencies and require intensive support and treatment prior to corrective surgery. The diagnosis is made on videofluoroscopy. The condition carries a good prognosis for cure with surgical myectomy of the cricopharyngeal muscle and the thyropharyngeal muscle, which make up the upper oesophageal sphincter. Temporary relief prior to surgery can be achieved by injection of the cricopharyngeal muscle with botulism toxin. Surgical treatment for dysphagia secondary to an underlying neurological, neuromuscular or pharyngeal weakness carries a guarded prognosis and will make aspiration pneumonia worse.


Dynamic state allocation for MEG source reconstruction.
Our understanding of the dynamics of neuronal activity in the human brain remains limited, due in part to a lack of adequate methods for reconstructing neuronal activity from noninvasive electrophysiological data. Here, we present a novel adaptive time-varying approach to source reconstruction that can be applied to magnetoencephalography (MEG) and electroencephalography (EEG) data. The method is underpinned by a Hidden Markov Model (HMM), which infers the points in time when particular states re-occur in the sensor space data. HMM inference finds short-lived states on the scale of 100ms. Intriguingly, this is on the same timescale as EEG microstates. The resulting state time courses can be used to intelligently pool data over these distinct and short-lived periods in time. This is used to compute time-varying data covariance matrices for use in beamforming, resulting in a source reconstruction approach that can tune its spatial filtering properties to those required at different points in time. Proof of principle is demonstrated with simulated data, and we demonstrate improvements when the method is applied to MEG.


The contribution of policy, law, management, research, and advocacy failings to the recent extinctions of three Australian vertebrate species.
Extinctions typically have ecological drivers, such as habitat loss. However, extinction events are also influenced by policy and management settings that may be antithetical to biodiversity conservation, inadequate to prevent extinction, insufficiently resourced, or poorly implemented. Three endemic Australian vertebrate species-the Christmas Island pipistrelle (Pipistrellus murrayi), Bramble Cay melomys (Melomys rubicola), and Christmas Island forest skink (Emoia nativitatis)-became extinct from 2009 to 2014. All 3 extinctions were predictable and probably preventable. We sought to identify the policy, management, research, and other shortcomings that contributed to their extinctions or failed to prevent them. These included a lack within national environmental legislation and policy of explicit commitment to the prevention of avoidable extinctions, lack of explicit accountability, inadequate resources for conservation (particularly for species not considered charismatic or not of high taxonomic distinctiveness), inadequate biosecurity, a slow and inadequate process for listing species as threatened, recovery planning that failed to consider the need for emergency response, inability of researchers to identify major threatening factors, lack of public engagement and involvement in conservation decisions, and limited advocacy. From these 3 cases, we recommend: environmental policy explicitly seeks to prevent extinction of any species and provides a clear chain of accountability and an explicit requirement for public inquiry following any extinction; implementation of a timely and comprehensive process for listing species as threatened and for recovery planning; reservation alone not be assumed sufficient to maintain species; enhancement of biosecurity measures; allocation of sufficient resources to undertake actions necessary to prevent extinction; monitoring be considered a pivotal component of the conservation response; research provides timely identification of factors responsible for decline and of the risk of extinction; effective dissemination of research results; advocacy by an informed public for the recovery of threatened species; and public involvement in governance of the recovery process. These recommendations should be applicable broadly to reduce the likelihood and incidence of extinctions.


Mechanistic study on the palladium(II)-catalyzed synthesis of 2,3-disubstituted indoles under aerobic conditions: anion effects and the development of a low-catalyst-loading process.
As a result of detailed mechanistic and kinetic studies, we have proposed that PdX2-catalyzed oxidative coupling of o-alkynylanilines 1 with terminal alkynes 2 under aerobic conditions is initiated by aminopalladation of 1 followed by ligand exchange of the resulting σ-indolylpalladium(II) complex with 2, reductive elimination and N-demethylation. Side reactions associated with intermediates on the way to 2,3-disubstituted indoles 3 were identified, and the roles of acetate and iodide in channeling the reaction towards the desired product were established. Based on kinetic and spectroscopic studies, the soluble iodide-ligated Pd(0) species was proposed to be the resting state of the catalyst and its oxidation to active Pd(II) species was the turnover-limiting step. Catalytic conditions with low loading of Pd(OAc)2 (0.0005 to 0.001 equiv) were subsequently developed.


Comparative immunological and biochemical analyses of viruses in the Venezuelan equine encephalitis complex.
Unclassified Venezuelan equine encephalitis (VEE) viruses Tonate (TON), Bijou Bridge (BB), Paramana (PARA), 71D-1252 and Cabassou (CAB) were characterized serologically and biochemically. The envelope glycoproteins of these and nine other VEE viruses representing VEE subtype variants I-AB, I-C, I-D, I-E, II, III and IV were separated by column isoelectric focusing. The E1 and E2 glycoproteins of all the Zwittergent-dissociated VEE viruses focused at pI 6.3 to 6.9 and pI 8.6 to 9.3 respectively. Haemagglutination-inhibition and neutralization tests using rabbit sera to the E2 glycoprotein of TON, BB and PARA viruses showed them to be indistinguishable from each other and closely related to prototype subtype III virus Mucambo (MUC). VEE strain 71D-1252 was also serologically closely related to prototype MUC virus. We proposed that MUC, TON and 71D-1252 VEE viruses be classified subtype III viruses, designated variants III-A, III-B and III-C respectively. CAB virus, which is not closely related to other VEE isolates, may represent a new VEE subtype (V). SDS-PAGE resolved the capsid protein (35 to 36 kdal) and two major envelope glycoproteins of 50 to 51 kdal (E1) and 51 to 58 kdal (E2) for all VEE viruses except CAB; the two glycoproteins of CAB virus co-migrated by PAGE with apparent identical mol. wt. of 51 kdal. Limited digestion of SDS-dissociated virus proteins with Staphylococcus aureus V8 protease produced identical peptide maps for serologically indistinguishable viruses. Oligonucleotide fingerprinting of virus RNA supported the close serological relationships observed at the genome level.


Real-time detection of H5N1 influenza virus through hyperbranched rolling circle amplification.
An isothermal amplification method was developed for the sensitive detection of the H5N1 influenza virus. The padlock probe specifically bound to the H5N1 target and circularized with T4 DNA ligase enzyme. Then this circular probe was amplified by hyperbranched rolling circle amplification (HRCA) using Phi29 DNA polymerase. The fluorescence intensity was recorded at different intervals by intercalation of SYBR green molecules into the double-stranded product of the HRCA reaction. At an optimum time of 88 min, a calibration plot with fine linearity was obtained. Using HRCA based on a padlock probe and Phi29 DNA polymerase, high selectivity and sensitivity were achieved. The biosensor response was linear toward H5N1 in the concentration range from 10 fM to 0.25 pM, with a detection limit of 9 fM at a signal/noise ratio of 3. By replacing the heat shock with pH shock, not only was the procedure for detection of H5N1 influenza simplified, but also the DNA molecules were protected from possible breaking at high temperature.


Pharmacological inhibition of arachidonate 15-lipoxygenase protects human spermatozoa against oxidative stress.
One of the leading causes of male infertility is defective sperm function, a pathology that commonly arises from oxidative stress in the germline. Lipid peroxidation events in the sperm plasma membrane result in the generation of cytotoxic aldehydes such as 4-hydroxynonenal (4HNE), which accentuate the production of reactive oxygen species (ROS) and cause cellular damage. One of the key enzymes involved in the metabolism of polyunsaturated fatty acids to 4HNE in somatic cells is arachidonate 15-lipoxygenase (ALOX15). Although ALOX15 has yet to be characterized in human spermatozoa, our previous studies have revealed a strong link between ALOX15 activity and the levels of oxidative stress and 4HNE in mouse germ cell models. In view of these data, we sought to assess the function of ALOX15 in mature human spermatozoa and determine whether the pharmacological inhibition of this enzyme could influence the level of oxidative stress experienced by these cells. By driving oxidative stress in vitro with exogenous H2O2, our data reveal that 6,11-dihydro[1]benzothiopyrano[4,3-b]indole (PD146176; a selective ALOX15 inhibitor) was able to significantly reduce several deleterious, oxidative insults in spermatozoa. Indeed, PD146176 attenuated the production of ROS, as well as membrane lipid peroxidation and 4HNE production in human spermatozoa. Accordingly, ALOX15 inhibition also protected the functional competence of these cells to acrosome react and bind homologous human zonae pellucidae. Together, these results implicate ALOX15 in the propagation of oxidative stress cascades within human spermatozoa and offer insight into potential therapeutic avenues to address male in fertility that arises from oxidative stress.


An area and power-efficient analog li-ion battery charger circuit.
The demand for greater battery life in low-power consumer electronics and implantable medical devices presents a need for improved energy efficiency in the management of small rechargeable cells. This paper describes an ultra-compact analog lithium-ion (Li-ion) battery charger with high energy efficiency. The charger presented here utilizes the tanh basis function of a subthreshold operational transconductance amplifier to smoothly transition between constant-current and constant-voltage charging regimes without the need for additional area- and power-consuming control circuitry. Current-domain circuitry for end-of-charge detection negates the need for precision-sense resistors in either the charging path or control loop. We show theoretically and experimentally that the low-frequency pole-zero nature of most battery impedances leads to inherent stability of the analog control loop. The circuit was fabricated in an AMI 0.5-μm complementary metal-oxide semiconductor process, and achieves 89.7% average power efficiency and an end voltage accuracy of 99.9% relative to the desired target 4.2 V, while consuming 0.16 mm(2) of chip area. To date and to the best of our knowledge, this design represents the most area-efficient and most energy-efficient battery charger circuit reported in the literature.


From NPC therapeutic target identification to potential treatment strategy.
Nasopharyngeal carcinoma (NPC) is relatively rare in Western countries but is a common cancer in southern Asia. Many differentially expressed genes have been linked to NPC; however, how to prioritize therapeutic targets and potential drugs from unsorted gene lists remains largely unknown. We first collected 558 upregulated and 993 downregulated NPC genes from published microarray data and the primary literatures. We then postulated that conversion of gene signatures into the protein-protein interaction network and analyzing the network topologically could provide insight into key regulators involved in tumorigenesis of NPC. Of particular interest was the presence of cliques, called fully connected subgraphs, in the inferred NPC networks. These clique-based hubs, connecting with more than three queries and ranked higher than other nodes in the NPC protein-protein interaction network, were further narrowed down by pathway analysis to retrieve 24 upregulated and 6 downregulated bottleneck genes for predicting NPC carcinogenesis. Moreover, additional oncogenes, tumor suppressor genes, genes involved in protein complexes, and genes obtained after functional profiling were merged with the bottleneck genes to form the final gene signature of 38 upregulated and 10 downregulated genes. We used the initial and final NPC gene signatures to query the Connectivity Map, respectively, and found that target reduction through our pipeline could efficiently uncover potential drugs with cytotoxicity to NPC cancer cells. An integrative Web site (http://126.96.36.199:8080/NPC) was established to facilitate future NPC research. This in silico approach, from target prioritization to potential drugs identification, might be an effective method for various cancer researches.


Validating the measurement of executive functions in an occupational context for adults with attention deficit hyperactivity disorder.
OBJECTIVE. The objectives of this study were to better understand the cognitive profile of adults with attention deficit hyperactivity disorder (ADHD), their occupational performance, and their quality of life (QoL) and to examine the validity of a cognitive-functional evaluation (CFE) battery for these adults. METHOD. Eighty-one adults with ADHD and 58 without ADHD completed ADHD symptom ratings, the Behavior Rating Inventory of Executive Function-Adult Version, and the Adult ADHD Quality-of-Life Scale. An occupational performance interview was administered to the ADHD group. RESULTS. A broad range of occupational concerns were reported. We found significant differences between groups on all measures. In the ADHD group, we found medium significant correlations among the measures. CONCLUSION. Adults with ADHD experience cognitive and functional difficulties in their daily lives associated with QoL. The results support the use of a CFE battery that has been shown to be sensitive and specific for these adults.


COVID-19 and Tinnitus.
Coronavirus disease 2019 (COVID-19) is an infectious respiratory disease caused by the severe acute respiratory syndrome coronavirus 2, brings with it a plethora of health concerns. Although most people have mild symptoms, which are respiratory in nature, some experience neurological symptoms, central nervous system manifestations, peripheral nervous manifestations, and skeletal muscle manifestations. But the damaging impact of COVID-19 virus on the hearing organs in the inner ear is a new finding yet to be explored. Currently, there is little evidence published connecting novel coronavirus and tinnitus directly. But according to the American Tinnitus Association, preexisting behavioral conditions make it more likely for patients to experience tinnitus due to the stress and depression associated with social isolation and infection avoidance. Hearing loss and Tinnitus is a common pathology seen in otolaryngology and there are numerous papers in literature describing its associations with other infections. However, this is the first reported case of hearing loss and tinnitus in a COVID-19 patient, in the State of Qatar, and this case report strives to contribute to the ocean of literature highlighting the need for otorhinolaryngologists to be aware of its correlation with COVID-19 virus.


LuxS: its role in central metabolism and the in vitro synthesis of 4-hydroxy-5-methyl-3(2H)-furanone.
Many bacteria produce extracellular molecules which function in cell-to-cell communication. One of these molecules, autoinducer 2 (AI-2), was first described as an extracellular signal produced by Vibrio harveyi to control luciferase expression. Subsequently, a number of bacteria have been shown to possess AI-2 activity in their culture supernatants, and bear the luxS gene product, which is required for AI-2 synthesis. In Porphyromonas gingivalis, luxS and pfs, encoding a 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTA/SAH'ase), form an operon, suggesting that S-adenosylhomocysteine (SAH) or 5'-methylthioadenosine (MTA) serves as a substrate for AI-2 production. Cell-free extracts of Escherichia coli MG1655, but not DH5alpha (which carries a luxS frame-shift mutation) were capable of generating AI-2 activity upon addition of SAH, but not MTA. S-Ribosyl-homocysteine (RH) derived from SAH also served as a substrate in E. coli MG1655 extracts. RH-supplemented cell-free extracts of Pseudomonas aeruginosa, a bacterium that lacks luxS, only generated AI-2 activity following the introduction of a plasmid containing the Por. gingivalis pfs-luxS operon. In addition, defined in vitro systems consisting of the purified LuxS proteins from Por. gingivalis, E. coli, Neisseria meningitidis or Staphylococcus aureus converted RH to homocysteine and a compound that exhibits AI-2 activity.4-Hydroxy-5-methyl-3(2H)-furanone was identified by mass spectrometry analysis as a major product formed in this in vitro reaction. In E. coli MG1655, expression of T3SH [the bacteriophage T3 S-adenosylmethionine (SAM) hydrolase] significantly reduced AI-2 activity in culture supernatants, suggesting that AI-2 production is limited by the amount of SAH produced in SAM-dependent transmethylase reactions. The authors suggest that the LuxS protein has an important metabolic function in the recycling of SAH. They also show that Ps. aeruginosa is capable of removing AI-2 activity, implying that this molecule may act as a nutrient. In many bacteria AI-2 may in fact represent not a signal molecule but a metabolite which is released early and metabolized in the later stages of growth.


Magnetic Cytoskeleton Affinity Purification of Microtubule Motors Conjugated to Quantum Dots.
We develop magnetic cytoskeleton affinity (MiCA) purification, which allows for rapid isolation of molecular motors conjugated to large multivalent quantum dots, in miniscule quantities, which is especially useful for single-molecule applications. When purifying labeled molecular motors, an excess of fluorophores or labels is usually used. However, large labels tend to sediment during the centrifugation step of microtubule affinity purification, a traditionally powerful technique for motor purification. This is solved with MiCA, and purification time is cut from 2 h to 20 min, a significant time-savings when it needs to be done daily. For kinesin, MiCA works with as little as 0.6 μg protein, with yield of ∼27%, compared to 41% with traditional purification. We show the utility of MiCA purification in a force-gliding assay with kinesin, allowing, for the first time, simultaneous determination of whether the force from each motor in a multiple-motor system drives or hinders microtubule movement. Furthermore, we demonstrate rapid purification of just 30 ng dynein-dynactin-BICD2N-QD (DDB-QD), ordinarily a difficult protein-complex to purify.


Mothers and Fathers in NICU: The Impact of Preterm Birth on Parental Distress.
Preterm birth is a stressful event for families. In particular, the unexpectedly early delivery may cause negative feelings in mothers and fathers. The aim of this study was to examine the relationship between preterm birth, parental stress and negative feelings, and the environmental setting of NICU. 21 mothers (age = 36.00 ± 6.85) and 19 fathers (age = 34.92 ± 4.58) of preterm infants (GA = 30.96 ± 2.97) and 20 mothers (age = 40.08 ± 4.76) and 20 fathers (age = 40.32 ± 6.77) of full-term infants (GA = 39.19 ± 1.42) were involved. All parents filled out the Parental Stressor Scale: Neonatal Intensive Care Unit, the Impact of Event Scale Revised, Profile of Mood States, the Multidimensional Scale of Perceived Social Support and the Post-Partum Bonding Questionnaire. Our data showed differences in emotional reactions between preterm and full-term parents. Results also revealed significant differences between mothers and fathers' responses to preterm birth in terms of stress, negative feelings, and perceptions of social support. A correlation between negative conditions at birth (e.g., birth weight and Neonatal Intensive Care Unit stay) and higher scores in some scales of Impact of Event Scale Revised, Profile of Mood States and Post-Partum Bonding Questionnaire were found. Neonatal Intensive Care Unit may be a stressful place both for mothers and fathers. It might be useful to plan, as soon as possible, interventions to help parents through the experience of the premature birth of their child and to begin an immediately adaptive mode of care.


Observation, analysis, and reconstruction of a twinned rainbow.
A photograph of a twinned rainbow, obtained on 11 May, 2012, in Dresden, Germany, is precisely calibrated with respect to lens projection and camera orientation. Since the twinning was only located in a part of the picture, it was possible to read out the red-green-blue intensity data from both a twinned and nontwinned position of the rainbow. These data were fitted with modeled spectra for polydisperse drop distributions, which were calculated with a Debye series algorithm and shifted in the scattering angle to account for the nonspherical shape of natural raindrops. The shift data were acquired from raytracing through realistic raindrop shapes modeled by two conjoined half-spheroids of different oblateness. Effective drop size distributions along the line of sight are derived from the fit for the two sampling positions and used to generate a true-color simulation of the original photograph. By this, the optical determination of physical rainfall properties is demonstrated.


Direct online HPLC-CV-AFS method for traces of methylmercury without derivatisation: a matrix-independent method for urine, sediment and biological tissue samples.
Mercury (Hg) is a global pollutant which occurs in different species, with methylmercury (MeHg) being the critical compound due to its neurotoxicity and bioaccumulation through the food chain. Methods for trace speciation of MeHg are therefore needed for a vast range of sample matrices, such as biological tissues, fluids, soils or sediments. We have previously developed an ultra-trace speciation method for methylmercury in water, based on a preconcentration HPLC cold vapour atomic fluorescence spectrometry (HPLC-CV-AFS) method. The focus of this work is mercury speciation in a variety of sample matrices to assess the versatility of the method. Certified reference materials were used where possible, and samples were spiked where reference materials were not available, e.g. human urine. Solid samples were submitted for commonly used digestion or extraction processes to obtain a liquid sample for injection into the analytical system. For MeHg in sediment samples, an extraction procedure was adapted to accommodate MeHg separation from high amounts of Hg(2+) to avoid an overload of the column. The recovery for MeHg determination was found to be in the range of 88-104% in fish reference materials (DOLT-2, DOLT-4, DORM-3), lobster (TORT-2), seaweed (IAEA-140/TM), sediments (ERM(®)-CC580) and spiked urine and has been proven to be robust, reliable, virtually matrix-independent and relatively cost-effective. Applications in the ultra-trace concentration range are possible using the preconcentration up to 200 mL, while for higher MeHg-containing samples, lower volumes can be applied. A comparison was carried out between species-specific isotope dilution gas chromatography inductively coupled plasma mass spectrometry (SSID-GC-ICP-MS) as the gold standard and HPLC-CV-AFS for biological tissues (liver, kidney and muscle of pilot whales), showing a slope of 1.008 and R (2) = 0.97, which indicates that the HPLC-CV-AFS method achieves well-correlated results for MeHg in biological tissues.


An efficient strategy for cell-based antibody library selection using an integrated vector system.
Cell panning of phage-displayed antibody library is a powerful tool for the development of therapeutic and imaging agents since disease-related cell surface proteins in native complex conformation can be directly targeted. Here, we employed a strategy taking advantage of an integrated vector system which allows rapid conversion of scFv-displaying phage into scFv-Fc format for efficient cell-based scFv library selection on a tetraspanin protein, CD9. A mouse scFv library constructed by using a phagemid vector, pDR-D1 was subjected to cell panning against stable CD9 transfectant, and the scFv repertoire from the enriched phage pool was directly transferred to a mammalian cassette vector, pDR-OriP-Fc1. The resulting constructs enabled transient expression of enough amounts of scFv-Fcs in HEK293E cells, and flow cytometric screening of binders for CD9 transfectant could be performed simply by using the culture supernatants. All three clones selected from the screening showed correct CD9-specificity. They could immunoprecipitate CD9 molecules out of the transfectant cell lysate and correctly stain endogenous CD9 expression on cancer cell membrane. Furthermore, competition assay with a known anti-CD9 monoclonal antibody (mAb) suggested that the binding epitopes of some of them overlap with that of the mAb which resides within the large extracellular loop of CD9. This study demonstrates that scFv-Fc from mammalian transient expression can be chosen as a reliable format for rapid screening and validation in cell-based scFv library selection, and the strategy described here will be applicable to efficient discovery of antibodies to diverse cell-surface targets.


Prostaglandins and the control of blood flow in the canine myocardium.
A comprehensive study was undertaken to evaluate the effects of inhibition of prostaglandin (PG) synthesis on a variety of reactions in the coronary vascular bed of anesthetized, open-chest dogs. In 23 dogs an electromagnetic flow probe (EMFP) and hydraulic occluder were placed around either the left anterior descending or circumflex branches of the coronary artery and a needle was inserted distral to the EMFP. Injections into the coronary artery of arachidonic acid (AA), bradykinin, adenosine, angiotensin, and PGE2 were given before and after inhibition of PG synthesis by indomethacin (IND) or meclofenamate (MF). The effects of the inhibitors on reactive hyperemia resulting from 5-, 10-, 15-, and 20-second occlusions and the dilation resulting from 90-second exposure to 8% O2 were also examined. In each experiment, inhibition of PG synthesis was ascertained by the elimination of vasodilation to AA. After administration of IND or MF, while baseline coronary blood flow was slightly reduced, the total increment of blood flow to vasodilator agents was not significantly altered. Whereas the peak dilation and volume of reactive hyperemia were decreased, the percent flow debt repaid was unchanged and total increment of coronary flow due to hypoxia-induced vasodilation was not significantly modified. Vasoconstrictor responses to angiotensin were also unchanged. These results indicate that while inhibitors of PG synthesis increase coronary resistance, they do not adversely affect vascular responsiveness. We conclude that prostaglandins play little, if any, role in modulating coronary blood flow.


The role of macrophages in the generation of T helper cells. V. Evidence for differential activation of short-lived T1 and long-lived T2 lymphocytes by the macrophage factors GRF and NMF.
The generation of T helper cells in vitro requires macrophages or macrophage-derived factors such as genetically related macrophage factor (GRF) or nonspecific macrophage factor (NMF). However, there is a basic difference of T helper cell induction when using particulate antigens. The present study demonstrates that this difference is based on the activation of two different T cell subsets. GRF activates short-lived 'T1' cells which amplify the induction of T2 cells, which are the helper cell precursors. Thus, the genetic restriction of T helper cell induction seen with soluble antigen or GRF lies on the level of macrophage or GRF interaction with T1 cells. NMF (or macrophages) and particulate antigens directly activate the helper cell precursor (T2) indicating no requirement for T1-T2 cooperation. The direct activation of the helper cell precursor with particulate antigens does not require histocompatible macrophages or NMF from histocompatible macrophages. The present results may explain some of the discrepancies reported in the literature concerning the genetic requirements and specificity of T cell activation.


Fundamental limitations for electroluminescence in organic dual-gate field-effect transistors.
A dual-gate organic field-effect transistor is investigated for electrically pumped lasing. The two gates can independently accumulate electrons and holes, yielding current densities exceeding the lasing threshold. Here, the aim is to force the electrons and holes to recombine by confining the charges in a single semiconducting film. It is found that independent hole and electron accumulation is mutually exclusive with vertical recombination and light emission.


Spontaneous subarachnoid hemorrhage of unknown origin: hospital course and long-term clinical and angiographic follow-up.
Hemorrhagic origin is unidentifiable in 10%-20% of patients presenting with spontaneous subarachnoid hemorrhage (SAH). While the patients in such cases do well clinically, there is a lack of long-term angiographic followup. The authors of the present study evaluated the long-term clinical and angiographic follow-up of a patient cohort with SAH of unknown origin that had been enrolled in the Barrow Ruptured Aneurysm Trial (BRAT). The BRAT database was searched for patients with SAH of unknown origin despite having undergone two or more angiographic studies as well as MRI of the brain and cervical spine. Follow-up was available at 6 months and 1 and 3 years after treatment. Analysis included demographic details, clinical outcome (Glasgow Outcome Scale, modified Rankin Scale [mRS]), and repeat vascular imaging. Subarachnoid hemorrhage of unknown etiology was identified in 57 (11.9%) of the 472 patients enrolled in the BRAT study between March 2003 and January 2007. The mean age for this group was 51 years, and 40 members (70%) of the group were female. Sixteen of 56 patients (28.6%) required placement of an external ventricular drain for hydrocephalus, and 4 of these subsequently required a ventriculoperitoneal shunt. Delayed cerebral ischemia occurred in 4 patients (7%), leading to stroke in one of them. There were no rebleeding events. Eleven patients were lost to followup, and one patient died of unrelated causes. At the 3-year follow-up, 4 (9.1%) of 44 patients had a poor outcome (mRS > 2), and neurovascular imaging, which was available in 33 patients, was negative. Hydrocephalus and delayed cerebral ischemia, while infrequent, do occur in SAH of unknown origin. Long-term neurological outcomes are generally good. A thorough evaluation to rule out an etiology of hemorrhage is necessary; however, imaging beyond 6 weeks from ictus has little utility, and rebleeding is unexpected.


Local factors associated with on-host flea distributions on prairie dog colonies.
Outbreaks of plague, a flea-vectored bacterial disease, occur periodically in prairie dog populations in the western United States. In order to understand the conditions that are conducive to plague outbreaks and potentially predict spatial and temporal variations in risk, it is important to understand the factors associated with flea abundance and distribution that may lead to plague outbreaks. We collected and identified 20,041 fleas from 6,542 individual prairie dogs of four different species over a 4-year period along a latitudinal gradient from Texas to Montana. We assessed local climate and other factors associated with flea prevalence and abundance, as well as the incidence of plague outbreaks. Oropsylla hirsuta, a prairie dog specialist flea, and Pulex simulans, a generalist flea species, were the most common fleas found on our pairs. High elevation pairs in Wyoming and Utah had distinct flea communities compared with the rest of the study pairs. The incidence of prairie dogs with Yersinia pestis detections in fleas was low (n = 64 prairie dogs with positive fleas out of 5,024 samples from 4,218 individual prairie dogs). The results of our regression models indicate that many factors are associated with the presence of fleas. In general, flea abundance (number of fleas on hosts) is higher during plague outbreaks, lower when prairie dogs are more abundant, and reaches peak levels when climate and weather variables are at intermediate levels. Changing climate conditions will likely affect aspects of both flea and host communities, including population densities and species composition, which may lead to changes in plague dynamics. Our results support the hypothesis that local conditions, including host, vector, and environmental factors, influence the likelihood of plague outbreaks, and that predicting changes to plague dynamics under climate change scenarios will have to consider both host and vector responses to local factors.


Porcine cysticercosis in southeast Uganda: seroprevalence in kamuli and kaliro districts.
The recent recognition of neurocysticercosis as a major cause of epilepsy in Uganda and changes in pig demography have lead to a need to better understand the basic epidemiology of Taenia solium infections in pigs and humans. Human exposure is a function of the size of the animal reservoir of this zoonosis. This is the first field survey for porcine cysticercosis to investigate the prevalence of antigen-positive pigs across an entire rural district of south-east Uganda. In our field surveys, 8.6% of 480 pigs screened were seropositive for the parasite by B158/B60 Ag-ELISA. In addition, of the 528 homesteads surveyed 138 (26%) did not have pit latrines indicating a high probability of pigs having access to human faeces and thus T. solium eggs. This study thus indicates the need for better data on this neglected zoonotic disease in Uganda, with a particular emphasis on the risk factors for infection in both pigs and humans. In this regard, further surveys of pigs, seroprevalence surveys in humans and an understanding of cysticercosis-related epilepsy are required, together with risk-factor studies for human and porcine infections.


Non-muscle myosin II deletion in the developing kidney causes ureter-bladder misconnection and apical extrusion of the nephric duct lineage epithelia.
In kidney development, connection of the nephric duct (ND) to the cloaca and subsequent sprouting of the ureteric bud (UB) from the ND are important for urinary exit tract formation. Although the roles of Ret signaling are well established, it remains unclear how intracellular cytoskeletal proteins regulate these morphogenetic processes. Myh9 and Myh10 encode two different non-muscle myosin II heavy chains, and Myh9 mutations in humans are implicated in congenital kidney diseases. Here we report that ND/UB lineage-specific deletion of Myh9/Myh10 in mice caused severe hydroureter/hydronephrosis at birth. At mid-gestation, the mutant ND/UB epithelia exhibited aberrant basal protrusion and ectopic UB formation, which likely led to misconnection of the ureter to the bladder. In addition, the mutant epithelia exhibited apical extrusion followed by massive apoptosis in the lumen, which could be explained by reduced apical constriction and intercellular adhesion mediated by E-cadherin. These phenotypes were not ameliorated by genetic reduction of the tyrosine kinase receptor Ret. In contrast, ERK was activated in the mutant cells and its chemical inhibition partially ameliorated the phenotypes. Thus, myosin II is essential for maintaining the apicobasal integrity of the developing kidney epithelia independently of Ret signaling.


Biochemical markers of bone turnover and bone loss at the lumbar spine and femoral neck: the Taiji study.
The purpose of this study was to ascertain whether biochemical markers of bone turnover predict bone loss. The survey was carried out in Taiji, Wakayama Prefecture, Japan. From a list of inhabitants aged 40-79 years, 400 participants (50 men and 50 women in each of four age groups) were selected randomly. Bone mineral density (BMD) was measured, and blood and urine samples of all participants were examined to obtain values for eight biochemical markers: alkaline phosphatase (ALP), bone Gla protein (BGP), type I procollagen (carboxyterminal peptide of type I procollagen; PICP), cross-linked carboxyterminal telopeptide region of type I collagen (ICTP), and urinary excretion of calcium (Ca), phosphate (P), pyridinoline (Pyr), and deoxypyridinoline (D-Pyr). Each marker was evaluated as a predictor of the rate of bone change in lumbar spine and femoral neck BMD over a 3-year period. The value of Pyr was significantly related to the change of lumbar spine BMD in men (P = 0.009), and that of BGP was found to be significant in women (P = 0.045). By contrast, none of the bone markers significantly correlated with bone loss at the femoral neck. The coefficient of determination at the lumbar spine was 5% and 7% at the femoral neck only. We conclude that biochemical markers of bone turnover cannot predict bone loss rates in middle-aged or elderly Japanese men and women over a 3-year period with sufficient accuracy for use in clinical decision making.


Agreement Attraction and Impasse Aversion: Reasons for Selecting a Poor Deal Over No Deal at All.
In the present studies, we examined the positive value of agreement and the negative value of impasse. Participants chose to give up real value and sacrifice economic efficiency in order to attain an agreement outcome and avoid an impasse outcome. A personally disadvantageous option was selected significantly more often when it was labeled "Agreement" rather than "Option A," and a personally advantageous option was avoided significantly more often when it was labeled "Impasse" rather than "Option B." In a face-to-face negotiation, a substantial proportion of individuals reached an agreement that was inferior to their best alternative to agreement. We showed that the appeal of agreement and the aversion to impasse both contribute to this effect, yet the aversion to impasse is the stronger of the two motivations. These findings have important implications for negotiators.


The 5-hydroxytryptamine2 agonist, (+-)-1-(2,5-dimethoxy-4-bromophenyl)-2-aminopropane stimulates the hypothalamic-pituitary-adrenal (HPA) axis. II. Biochemical and physiological evidence for the development of tolerance after chronic administration.
In order to investigate the long term effects of the 5-hydroxytryptamine2 (5-HT2) receptor agonist, (+-)-1-(2,5-dimethoxy-4-bromophenyl)-2-aminopropane (DOB), on hypothalamic-pituitary-adrenal (HPA) axis activity, DOB (0.35 mg/kg/day) was administered via osmotic minipumps to rats for 7 days at which time plasma adrenocorticotrophic hormone (ACTH), corticosterone and regional brain corticotropin-releasing factor (CRF) concentrations were measured. In addition, anterior pituitary CRF and frontal cortical 5-HT2 receptor binding was measured. Seven-day infusion of DOB resulted in tolerance to the stimulatory actions of the drug on the HPA axis as evidenced by the return of plasma ACTH and corticosterone concentrations to base-line values. Moreover, rats treated chronically with DOB exhibited decreased numbers of both anterior pituitary CRF and cortical and hypothalamic 5-HT2 receptor. These receptor changes were physiologically significant as challenges doses of DOB or CRF resulted in blunted ACTH responses. Chronic DOB infusion was without effect on CRF concentrations in all hypothalamic and extrahypothalamic brain regions studied. A series of time course experiments revealed that DOB-induced increases in plasma corticosterone returned to base-line by 2-days postimplantation. This effect was apparently associated with down-regulation of the 5-HT2 receptor because high-affinity cortical [3H]DOB and hypothalamic (+-)-[125I]-1-(2,5-dimethoxy-4-iodophenyl)-2-amino-propane binding were decreased at this time as well. Although median eminence CRF content was unchanged at all time points, anterior pituitary CRF receptor binding was significantly decreased 7 days postimplantation.(ABSTRACT TRUNCATED AT 250 WORDS)


Human Hemoglobin Subunit Beta Functions as a Pleiotropic Regulator of RIG-I/MDA5-Mediated Antiviral Innate Immune Responses.
Hemoglobin is an important oxygen-carrying protein and plays crucial roles in establishing host resistance against pathogens and in regulating innate immune responses. The hemoglobin subunit beta (HB) is an essential component of hemoglobin, and we have previously demonstrated that the antiviral role of the porcine HB (pHB) is mediated by promoting type I interferon pathways. Thus, considering the high homology between human HB (hHB) and pHB, we hypothesized that hHB also plays an important role in the antiviral innate immunity. In this study, we characterized hHB as a regulatory factor for the replication of RNA viruses by differentially regulating the RIG-I- and MDA5-mediated antiviral signaling pathways. Furthermore, we showed that hHB directly inhibited MDA5-mediated signaling by reducing the MDA5-double-stranded RNA (dsRNA) interaction. Additionally, hHB required hHB-induced reactive oxygen species (ROS) to promote RIG-I-mediated signaling through enhancement of K63-linked RIG-I ubiquitination. Taken together, our findings suggest that hHB is a pleiotropic regulator of RIG-I/MDA5-mediated antiviral responses and further highlight the importance of the intercellular microenvironment, including the redox state, in regulating antiviral innate immune responses.IMPORTANCE Hemoglobin, the most important oxygen-carrying protein, is involved in the regulation of innate immune responses. We have previously reported that the porcine hemoglobin subunit beta (HB) exerts antiviral activity through regulation of type I interferon production. However, the antiviral activities and the underlying mechanisms of HBs originating from other animals have been poorly understood. Here, we identified human HB (hHB) as a pleiotropic regulator of the replication of RNA viruses through regulation of RIG-I/MDA5-mediated signaling pathways. hHB enhances RIG-I-mediated antiviral responses by promoting RIG-I ubiquitination depending on the hHB-induced reactive oxygen species (ROS), while it blocks MDA5-mediated antiviral signaling by suppressing the MDA5-dsRNA interaction. Our results contribute to an understanding of the crucial roles of hHB in the regulation of the RIG-I/MDA5-mediated signaling pathways. We also provide novel insight into the correlation of the intercellular redox state with the regulation of antiviral innate immunity.


Single oblique osteotomy for correction of antebrachial angular and torsional deformities in a dog.
CASE DESCRIPTION A 9-month-old 4.6-kg (10.1-lb) spayed female Shih Tzu was examined for a 4-week history of left forelimb lameness. CLINICAL FINDINGS Moderate left forelimb lameness was present when weight bearing, accompanied by buckling of the left carpal joint and moderate signs of pain with hyperextension of the elbow joint. A 32° angular deformity of the left radius in a 45° craniomedial oblique plane (equivalent to 23° valgus and 23° procurvatum) and a 55° external torsional deformity were measured on 3-D reconstructed CT images. Humeroulnar incongruity of the left elbow joint was also evident. Single oblique osteotomy (SOO) and dynamic proximal ulnar osteotomy were recommended. TREATMENT AND OUTCOME Creation of a 3-D printed bone model from reconstructed CT images aided surgical planning. The SOO was located 45° medial to the sagittal plane of the left radius at the level of the center of rotation of angulation. The SOO was oriented 32° distolaterally from the transverse plane of the radius and traversed the left ulna. The bones were rotated along the osteotomy into grossly proper alignment and stabilized with a plate and bone screws. Dynamic proximal ulnar osteotomy was then performed. Six months after surgery, radiographs showed remodeling at the SOO site and the lameness had resolved. The owners expressed satisfaction with the outcome. CLINICAL RELEVANCE In dogs with angular and torsional long bone deformities, SOO may be a viable alternative to wedge osteotomies. The SOO simultaneously addressed angular and torsional deformities without bone loss and provided rigid internal fixation.


Genomic-driven insights into changes in aging skin.
Like all tissues, the skin ages due to the passage of time (chronologic aging). However, skin is also exposed to external insults, such as sunlight. Aging due to chronic sun exposure (photoaging) is characterized clinically by wrinkling, dyspigmentation and other changes. Chronologic and photoaging of skin have been distinguished at the structural, cellular and molecular levels. However, many underlying mechanisms remain a mystery. Recent sequencing of the human genome and development of genome-wide microarray platforms now permit analysis of skin aging at the level of gene expression. Analysis of gene expression differences between young and older sun-protected and sun-exposed skin showed that photoaging produces many similar (but more severe) changes in gene expression versus chronologic aging. However, some changes are unique to one form of aging or the other. Bioinformatics tools also enable an integrated analysis of gene expression themes and pathways, which may provide new insights into the mechanisms of skin aging and possible interventions.