Histone deacetylase inhibition with valproic acid downregulates osteocalcin gene expression in human dental pulp stem cells and osteoblasts: evidence for HDAC2 involvement.
Adult mesenchymal stem cells, such as dental pulp stem cells, are of great interest for cell-based tissue engineering strategies because they can differentiate into a variety of tissue-specific cells, above all, into osteoblasts. In recent years, epigenetic studies on stem cells have indicated that specific histone alterations and modifying enzymes play essential roles in cell differentiation. However, although several studies have reported that valproic acid (VPA)-a selective inhibitor of histone deacetylases (HDAC)-enhances osteoblast differentiation, data on osteocalcin expression-a late-stage marker of differentiation-are limited. We therefore decided to study the effect of VPA on dental pulp stem cell differentiation. A low concentration of VPA did not reduce cell viability, proliferation, or cell cycle profile. However, it was sufficient to significantly enhance matrix mineralization by increasing osteopontin and bone sialoprotein expression. In contrast, osteocalcin levels were decreased, an effect induced at the transcriptional level, and were strongly correlated with inhibition of HDAC2. In fact, HDAC2 silencing with shRNA produced a similar effect to that of VPA treatment on the expression of osteoblast-related markers. We conclude that VPA does not induce terminal differentiation of osteoblasts, but stimulates the generation of less mature cells. Moreover, specific suppression of an individual HDAC by RNA interference could enhance only a single aspect of osteoblast differentiation, and thus produce selective effects.


Interpretation of oral fluid tests for drugs of abuse.
Oral fluid testing for drugs of abuse offers significant advantages over urine as a test matrix. Collection can be performed under direct observation with reduced risk of adulteration and substitution. Drugs generally appear in oral fluid by passive diffusion from blood, but also may be deposited in the oral cavity during oral, smoked, and intranasal administration. Drug metabolites also can be detected in oral fluid. Unlike urine testing, there may be a close correspondence between drug and metabolite concentrations in oral fluid and in blood. Interpretation of oral fluid results for drugs of abuse should be an iterative process whereby one considers the test results in the context of program requirements and a broad scientific knowledge of the many factors involved in determining test outcome. This review delineates many of the chemical and metabolic processes involved in the disposition of drugs and metabolites in oral fluid that are important to the appropriate interpretation of oral fluid tests. Chemical, metabolic, kinetic, and analytic parameters are summarized for selected drugs of abuse, and general guidelines are offered for understanding the significance of oral fluid tests.


Assessing Physiological Stress Responses to Word Retrieval in Individuals with Aphasia: A Preliminary Study.
Word retrieval difficulty can be distressing for individuals with aphasia. However, physiological stress responses to word retrieval are relatively unstudied. This study aims to measure the physiological response (i.e., heart rate and respiratory rate) and perceived stress in individuals with aphasia during a naming task to understand whether word retrieval may be a stressor to them. An aphasia group (n = 7) and a healthy group (n = 38) participated in 4 experimental conditions, including rest, counting, and high-frequency and low-frequency word conditions, while their physiological parameters were measured throughout the sessions. Stress ratings were also obtained for each condition. Significant differences in heart rate were found while participants were engaged in speech and/or language tasks, compared to the rest condition, for both groups. Significant differences in respiratory rate were found between the rest and low-frequency word conditions in the aphasia group. No significant difference was found in heart rate between 2 groups, but a difference in respiratory rate was found between 2 groups in the low-frequency word condition. There were some discrepancies between stress level and physiological parameters. Individuals with aphasia perceived greater stress in the naming tasks, however, the perceived stress was not necessarily parallel to the physiological parameters.


Randomized controlled trial of acupuncture versus sham acupuncture in autism spectrum disorder.
We aim to study the efficacy of acupuncture versus sham acupuncture in children with autism spectrum disorder. A single-blind randomized control trial was conducted in 50 children. These children were randomly assigned to the treatment group with tongue acupuncture (40 sessions over 8 weeks) or the control group (sham tongue acupuncture to nonacupoints in the tongue). There was improvement in both the treatment and control groups in all assessed measures but more so in the treatment than in the control group: (1) eye-hand coordination, performance, and practical reasoning of Griffiths Mental Developmental Scale; (2) sensory-motor, social, affectual, language, and total score of Ritvo-Freeman Real Life Scale; (3) Comprehension Language age in the Reynell Language Developmental Scale; and (4) Total Score and Mental Age in Symbolic Play Test. The only statistically significant improvement in the treatment as compared to the control group was seen in self-care and cognition domains of the Functional Independence Measure for children. We had demonstrated that a short course of acupuncture had efficacy in improving various developmental and behavioral aspects of children with autism. The long-term efficacy in functional gain needs to be further explored.


A model-based projection of historical state of a coastal ecosystem: Relevance of phytoplankton stoichiometry.
We employed a coupled physical-biogeochemical modelling framework for the reconstruction of the historic (H), pre-industrial state of a coastal system, the German Bight (southeastern North Sea), and we investigated its differences with the recent, control (C) state of the system. According to our findings: i) average winter concentrations of dissolved inorganic nitrogen and phosphorus (DIN and DIP) concentrations at the surface are ∼70-90% and ∼50-70% lower in the H state than in the C state within the nearshore waters, and differences gradually diminish towards off-shore waters; ii) differences in average growing season chlorophyll a (Chl) concentrations at the surface between the two states are mostly less than 50%; iii) in the off-shore areas, Chl concentrations in the deeper layers are affected less than in the surface layers; iv) reductions in phytoplankton carbon (C) biomass under the H state are weaker than those in Chl, due to the generally lower Chl:C ratios; v) in some areas the differences in growth rates between the two states are negligible, due to the compensation by lower light limitation under the H state, which in turn explains the lower Chl:C ratios; vi) zooplankton biomass, and hence the grazing pressure on phytoplankton is lower under the H state. This trophic decoupling is caused by the low nutritional quality (i.e., low N:C and P:C) of phytoplankton. These results call for increased attention to the relevance of the acclimation capacity and stoichiometric flexibility of phytoplankton for the prediction of their response to environmental change.


Effects of mineralocorticoids on transport properties of cortical collecting duct basolateral membrane.
The effects of long-term mineralocorticoid (deoxycorticosterone acetate, DOCA) elevation for 9-16 days on the active and passive transport properties of the basolateral cell membrane of rabbit cortical collecting ducts were assessed using microelectrode techniques. It was found that both the transepithelial membrane voltage (Vte) and basolateral membrane voltage (Vb) hyperpolarized and the basolateral membrane conductance (Gb) increased on chronic elevation of mineralocorticoid levels. Barium (5 mM) addition to the bathing solution effectively blocked an induced K+ current across the basolateral cell membrane without an immediate affect on the other barriers. Therefore Ba2+ was used to quantitate the basolateral cell membrane K+ conductance (GbK). It was found that GbK increased from 1.0 +/- 0.2 mS X cm-2 (controls) to 3.7 +/- 1.0 mS X cm-2 in DOCA-treated animals. The basolateral membrane electrogenic pump current (Ibact) was quantitated from the change in the basolateral membrane equivalent emf on addition of either ouabain (0.1 mM) to the bath or of amiloride (50 microM) to the perfusate. There was a large increase in Ibact from 32 microA X cm-2 in controls to 195 microA X cm-2 in the DOCA-treated group. In addition, the estimated Na+-to-K+ coupling ratio of the Na+ pump was observed to increase from 1.6 to 1.0 in the control group to 3.2 to 1.0 in the DOCA-treated group. The estimated basolateral membrane passive K+ current (IbK) increased from a value that was not significantly different from 0 in controls to approximately -45 microA X cm-2 (from bath to cell) in the DOCA-treated group. These findings support a model whereby mineralocorticoids induce an increase in electrogenic Na+ pump activity in response to chronically elevated rates of Na+ transport. This results in a hyperpolarization of Vb, which is well above EbK, thereby resulting in a net driving force for K+ uptake into the cell, bringing about an increased rate of K+ secretion.


[Risk factors negatively affecting on the formation of musculoskeletal system in children and adolescents in the present conditions].
Identifying risk factors affecting the formation of the musculoskeletal system (MSS) in children and adolescents is considered by the author as a necessary condition for the implementation of prevention, timely diagnosis and adequate correction of the MSS disorders and diseases. Introduction in the educational process developed by the author for the first time a conceptual model of prevention and correction of the MSS disorders and diseases in schoolchildren allowed significantly reduce the prevalence of functional disorders and early forms of the MSS diseases in students of a number of comprehensive schools in Moscow by 50%.


Evaluating the Synergistic Potency of Essential Oils and Antibiotics Against Klebsiella pneumoniae BLSE Strains.
Klebsiella pneumoniae producing extended-spectrum β-lactamases (ESBL) continues to pose huge therapeutic challenges in the treatment of infections, primarily urinary infections, due to its multidrug resistance to antibiotics. Therefore, there is a need for research on this topic to investigate ways to reduce the spread of antibiotic resistance, identify novel therapeutic approaches to treat these infections and gain a better understanding of the mechanisms of resistance. In this context, this study aimed to analyze the chemical composition of essential oils (EOs) of Thymus algeriensis, Syzygium aromaticum, and Eucalyptus globulus and assess their activity against K. pneumoniae ESBL strains, as well as the interaction type between these EOs and antibiotics used for the treatment of K. pneumoniae ESBL infections. The composition of the EOs was determined by gas chromatography-mass spectrometry (GC-MS). The activity of EOs was tested using the disc diffusion and liquid microdilution methods. The type of interaction between EOs and antibiotics was studied using the agar disk diffusion and chessboard methods. The analysis of the EO of T. algeriensis showed that the main compounds were thymol (23.14%), linalool (18.44%), and p-cymene (16.17%). The main constituents of EO of E. globulus were eucalyptol (54.29%), α-pinene (17.32%), aromadendrene (7.02%), and pinocarveol (6.32%). As for the EO of S. aromaticum, the major constituents were eugenol (80.46%) and eugenol acetate (16.23%). Results of the activity tests showed that all three EOs were active against the tested strains, with inhibition diameters ranging from 7.39±0.44mm to 32.4±1.05mm and minimum inhibitory concentrations (MICs) varying from 2 to 441.5±5.66 mg/ml. A synergistic interaction was obtained between amoxicillin-clavulanic acid and T. algeriensis EO against two strains of K. pneumoniae ESBL. These results demonstrate the potential of our EOs to inhibit multi-resistant pathogenic ESBL strains, as well as their synergistic interaction with antibiotics used in therapy, which could be an alternative to the use of antibiotics alone in treatment to fight against these multi-resistant pathogenic bacteria.


Guidance on left bundle branch pacing using continuous pacing technique and changes in lead V1 characteristics under real-time monitoring.
The changes in the morphology and characteristics of the V1 leads during left bundle branch capturing still need to be fully understood. This study aims to provide some suggestions about the LBB capture process through the morphology and characteristics of the V1 lead. LBBP using the continuous pacing and morphology monitoring technique during screw-in using a revolving connector (John Jiang's connecting cable). The morphology and features of V1 leads are recorded by continuous monitoring technology. The most common morphology in the LVSP stage is QR, while in the NS-LBBP (low output) stage and the NS-LBBP (lower output) stage, it is rSR. In the S-LBBP stage, it is rsR. The predominant morphology is with r/R waves in S-LBBP, which includes variations like rSR, rsR, rSr, rsr, rR, rs, rS, and R type, making up 96.7% of the total. The r waves in lead V1 are associated with agitated myocardium conducted from the left bundle branch. The initial r-wave in lead V1 may be a marker during the follow-up of patients with selective LBB capture.


Changes in avidity and level of immunoglobulin G antibodies to Mycobacterium tuberculosis in sera of patients undergoing treatment for pulmonary tuberculosis.
Much is known about specific antibodies and their titers in patients with tuberculosis. However, little is known about the avidity of these antibodies or whether changes in avidity occur during the progression of the disease or during treatment. The aims of this study were to determine the avidity of antibodies to Mycobacterium tuberculosis in patients with pulmonary tuberculosis, to explore the value of avidity determination for the diagnosis of tuberculosis, and to study changes in levels of antibodies and their avidity during treatment. Antibody avidity was measured by an enzyme-linked immunosorbent assay with thiocyanate elution. Avidity indices and serum levels of immunoglobulin G to M. tuberculosis were determined for 22 patients with pulmonary tuberculosis before and during treatment and for 24 patients with other pulmonary diseases. Antibody levels and avidity were both significantly higher in untreated tuberculosis patients than in the controls. Avidity determination had more diagnostic potential than determination of the antibody levels. Tuberculosis patients with a long duration of symptoms had higher antibody avidity than those with a recent onset of symptoms, indicating affinity maturation of specific antibodies during active disease. In the early phase of treatment, a decrease in antibody avidity was observed for 73% of all tuberculosis patients, accompanied by an initial increase in antibody levels in 36% of these patients. These phenomena could be explained by an intense stimulation of the humoral response by antigens released from killed bacteria, reflecting early bactericidal activity of antituberculous drugs leading to the production of low-affinity antibodies against these released antigens.


On the difference in the net ecosystem exchange of CO2 between deciduous and evergreen forests in the southeastern United States.
The southeastern United States is experiencing a rapid regional increase in the ratio of pine to deciduous forest ecosystems at the same time it is experiencing changes in climate. This study is focused on exploring how these shifts will affect the carbon sink capacity of southeastern US forests, which we show here are among the strongest carbon sinks in the continental United States. Using eight-year-long eddy covariance records collected above a hardwood deciduous forest (HW) and a pine plantation (PP) co-located in North Carolina, USA, we show that the net ecosystem exchange of CO2 (NEE) was more variable in PP, contributing to variability in the difference in NEE between the two sites (ΔNEE) at a range of timescales, including the interannual timescale. Because the variability in evapotranspiration (ET) was nearly identical across the two sites over a range of timescales, the factors that determined the variability in ΔNEE were dominated by those that tend to decouple NEE from ET. One such factor was water use efficiency, which changed dramatically in response to drought and also tended to increase monotonically in nondrought years (P < 0.001 in PP). Factors that vary over seasonal timescales were strong determinants of the NEE in the HW site; however, seasonality was less important in the PP site, where significant amounts of carbon were assimilated outside of the active season, representing an important advantage of evergreen trees in warm, temperate climates. Additional variability in the fluxes at long-time scales may be attributable to slowly evolving factors, including canopy structure and increases in dormant season air temperature. Taken together, study results suggest that the carbon sink in the southeastern United States may become more variable in the future, owing to a predicted increase in drought frequency and an increase in the fractional cover of southern pines.


Health Care Professionals' Experiences of Patient-Professional Communication Over Patient Portals: Systematic Review of Qualitative Studies.
The popularity of web-based patient-professional communication over patient portals is constantly increasing. Good patient-professional communication is a prerequisite for high-quality care and patient centeredness. Understanding health care professionals' experiences of web-based patient-professional communication is important as they play a key role in engaging patients to use portals. More information is needed on how patient-professional communication could be supported by patient portals in health care. This systematic review of qualitative studies aims to identify how health care professionals experience web-based patient-professional communication over the patient portals. Abstract and full-text reviews were conducted by 2 reviewers independently. A total of 4 databases were used for the study: CINAHL (EBSCO), ProQuest (ABI/INFORM), Scopus, and PubMed. The inclusion criteria for the reviewed studies were as follows: the examination of health care professionals' experiences, reciprocal communication between patients and health care professionals, peer-reviewed scientific articles, and studies published between 2010 and 2019. The Joanna Briggs Institute's quality assessment criteria were used in the review process. A total of 13 included studies were analyzed using a thematic synthesis, which was conducted by 3 reviewers. A total of 6 analytical themes concerning health care professionals' experiences of web-based patient-professional communication were identified. The themes were related to health care professionals' work, change in communication over patient portals, patients' use of patient portals, the suitability of patient portals for communication, the convenience of patient portals for communication, and change in roles. Health care professionals' experiences contain both positive and negative insights into web-based patient-professional communication over patient portals. Most commonly, the positive experiences seem to be related to the patients and patient outcomes, such as having better patient engagement. Health care professionals also have negative experiences, for example, web-based patient-professional communication sometimes has deficiencies and has a negative impact on their workload. These negative experiences may be explained by the poor functionality of the patient portals and insufficient training and resources. To reduce health care professionals' negative experiences of web-based patient-professional communication, their experiences should be taken into account by policy makers, health care organizations, and information technology enterprises when developing patient portals. In addition, more training regarding web-based patient-professional communication and patient portals should be provided to health care professionals.


Lottery incentives have short-term impact on ART initiation among men: results from a randomized pilot study.
Among people living with HIV in South Africa, viral suppression is lower among men than women. The study aim was to test the impact of lottery incentives, which reward positive health choice (e.g. antiretroviral therapy (ART) linkage) with a chance to win a prize, on strengthening the HIV care continuum including ART initiation and viral suppression for men. We conducted a randomized, prospective trial of lottery incentives in the context of HIV testing and linkage to ART in rural KwaZulu-Natal, South Africa. Men living with HIV were randomly allocated to: lottery incentives and motivational text messages or motivational text messages only. Lottery prize eligibility was conditional on clinic registration, ART initiation, or viral suppression by one, three and six months respectively. After completing each continuum step, participants in the lottery group were notified whether they had won and were encouraged to continue in care. Lottery prizes were either a mobile phone, data or a gift card (valued at R1000/$100). Kaplan-Meier curves were plotted to determine time to ART initiation by study group. The primary outcome was viral suppression at six months. Between November 2017 and December 2018, we tested 740 men for HIV and enrolled 131 HIV-positive men who reported not being on ART. At baseline, 100 (76%) participants were 30 years and older, 95 (73%) were unemployed and the median CD4 count was 472 cells/μL. At study exit, 84% (110/131) of participants had visited a clinic and 62% (81/131) were virally suppressed. Compared to motivational text messages, lottery incentives decreased the median time to ART initiation from 126 to 66 days (p = 0.0043, age-adjusted Cox regression) among all participants, and, from 134 days to 20 days (p = 0.0077) among participants who were not virally suppressed at baseline. Lottery incentives had an inconclusive effect on clinic registration (RR = 1.21, 95% CI: 0.83 to 1.76) and on viral suppression at six months (RR = 1.13, 95% CI: 0.73 to 1.75) compared to motivational text messages. Conditional lottery incentives shortened the time to ART initiation among South African men. Behavioural economics strategies strengthen linkage to ART, but the study power was limited to see an impact on viral suppression. NCT03808194.


Global cell surface conformational shift mediated by a Candida albicans adhesin.
Candida albicans maintains both commensal and pathogenic states in humans. Both states are dependent on cell surface-expressed adhesins, including those of the Als family. Heterologous expression of Als5p at the surface of Saccharomyces cerevisiae results in Als5p-mediated adhesion to various ligands, followed by formation of multicellular aggregates. Following adhesion of one region of the cell to fibronectin-coated beads, the entire surface of the cells became competent to mediate cell-cell aggregation. Aggregates formed in the presence of metabolic inhibitors or signal transduction inhibitors but were reduced in the presence of 8-anilino-1-naphthalene-sulfonic acid (ANS) or Congo Red (CR), perturbants that inhibit protein structural transitions. These perturbants also inhibited aggregation of C. albicans. An increase in ANS fluorescence, which accompanied Als-dependent cellular adhesion, indicated an increase in cell surface hydrophobicity. In addition, C. albicans and Als5p-expressing S. cerevisiae showed an aggregation-induced birefringence indicative of order on the cell surface. The increase in birefringence did not occur in the presence of the aggregation disruptants ANS and CR. These results suggest a model for Als5p-mediated aggregation in which an adhesion-triggered change in the conformation of Als5p propagates around the cell surface, forming ordered aggregation-competent regions.


NSAIDs suppress the expression of claudin-2 to promote invasion activity of cancer cells.
Non-steroidal anti-inflammatory drugs (NSAIDs) show chemopreventive effects; however, the precise molecular mechanism of these effects is still unclear. On the other hand, the expression of proteins that form tight junctions (TJs) (such as claudins) in clinically isolated tumors is frequently altered relative to normal tissue. We previously reported that NSAIDs upregulate the expression of claudin-4 and that this upregulation contributes to NSAID-dependent inhibition of both migration activity and anchorage-independent growth of cancer cells. In the current study, we have systematically examined the effects of various NSAIDs on the expression of various TJ proteins and have found that NSAIDs specifically and drastically inhibit the expression of claudin-2. Overexpression or suppression of claudin-2 expression caused stimulation or inhibition, respectively, of the invasion and migration activity of cancer cells. Furthermore, NSAIDs inhibited the invasion and migration activity of cancer cells and this inhibition was suppressed by overexpression of claudin-2. In contrast, neither cell growth nor apoptosis induced by lack of anchorage of cancer cells was affected by overexpression or suppression of expression of claudin-2. These results suggest that inhibition of claudin-2 expression by NSAIDs contributes to NSAID-dependent inhibition of invasion of cancer cells in vitro and that it may be involved in the chemopreventive effects of NSAIDs by inhibiting metastasis in vivo.


Volatile hydrocarbons inhibit methanogenic crude oil degradation.
Methanogenic degradation of crude oil in subsurface sediments occurs slowly, but without the need for exogenous electron acceptors, is sustained for long periods and has enormous economic and environmental consequences. Here we show that volatile hydrocarbons are inhibitory to methanogenic oil biodegradation by comparing degradation of an artificially weathered crude oil with volatile hydrocarbons removed, with the same oil that was not weathered. Volatile hydrocarbons (nC5-nC10, methylcyclohexane, benzene, toluene, and xylenes) were quantified in the headspace of microcosms. Aliphatic (n-alkanes nC12-nC34) and aromatic hydrocarbons (4-methylbiphenyl, 3-methylbiphenyl, 2-methylnaphthalene, 1-methylnaphthalene) were quantified in the total hydrocarbon fraction extracted from the microcosms. 16S rRNA genes from key microorganisms known to play an important role in methanogenic alkane degradation (Smithella and Methanomicrobiales) were quantified by quantitative PCR. Methane production from degradation of weathered oil in microcosms was rapid (1.1 ± 0.1 μmol CH4/g sediment/day) with stoichiometric yields consistent with degradation of heavier n-alkanes (nC12-nC34). For non-weathered oil, degradation rates in microcosms were significantly lower (0.4 ± 0.3 μmol CH4/g sediment/day). This indicated that volatile hydrocarbons present in the non-weathered oil inhibit, but do not completely halt, methanogenic alkane biodegradation. These findings are significant with respect to rates of biodegradation of crude oils with abundant volatile hydrocarbons in anoxic, sulphate-depleted subsurface environments, such as contaminated marine sediments which have been entrained below the sulfate-reduction zone, as well as crude oil biodegradation in petroleum reservoirs and contaminated aquifers.


Adsorption and enzymatic cleavage of osteopontin at interfaces with different surface chemistries.
Osteopontin is a highly charged glycoprotein present in the extra cellular matrix of a wide range of tissues. It is, in particular, relevant for biomaterials through its role in mineralized tissue remodeling. The adsorption and enzymatic cleavage of osteopontin at four different surface chemistries (methyl-, carboxylic-, and amine-terminated alkanethiol self-assembled monolayers and bare gold) have been studied utilizing a combination of the quartz crystal microbalance with dissipation and surface plasmon resonance. Full length bovine milk osteopontin was used which is well characterized with respect to post-translational modifications. Osteopontin adsorbed at all the surfaces formed thin (approximately 2-5 nm) hydrated layers with the highest amount of protein and the highest density layers observed at the hydrophobic surface. Less protein and a higher level of hydration was observed at the polar surfaces with the highest level of hydration being observed at the gold surface. The energy dissipation of these thin films (as measured by the DeltaD/DeltaF value) was altered at the different surface chemistries and interestingly a higher dissipation correlated with a higher density. Thrombin was able to bind and cleave the surface bound osteopontin at the hydrophobic surface. The altered levels of osteopontin binding, hydration of the layer, and susceptibility to thrombin cleavage suggest that osteopontin adopts different conformations and/or orientations at the different material surfaces.


A manufactu(RED) ethics: labor, HIV, and the body in Lesotho's "sweat-free" garment industry.
Employing mostly women and producing for major U.S. labels, Lesotho's primarily foreign-owned garment industry undertook efforts to become "sweat-free" in 2006; simultaneously, it also began producing for the Product(RED) campaign. This article explores the parameters and ethical challenges of an industry-wide, public-private partnership providing HIV prevention and treatment services in this industry. Here, HIV services are intimately bound up in emerging patterns of humanitarian consumption and the production of an ethical industry. Within this ethical production zone, all is not what it seems: Labor violations persist, workers confront occupational hazards, and an elaborate theatrics of ethical practice plays out on the factory floor during routine inspections. This article explores the place and purpose of HIV treatment in the context of such humanitarian fetishism, highlighting the uses to which worker bodies are put and the conceptions of bodily well-being that prevail in these new "moral" economies.


Influence of dietary crude protein and phosphorus on ileal digestion of phosphorus and amino acids in growing pigs.
A study was conducted to investigate the effect of dietary CP levels on ileal P digestion in growing pigs. A total of 18 ileal-cannulated pigs (44.2 ± 3.2 kg initial BW) were used in a duplicated 9 × 3 incomplete Latin square design, with 9 treatments and three 7-d experimental periods giving 6 replicates per treatment. The 9 treatments consisted of 1 N-free diet to estimate basal endogenous loss of AA and 8 corn-soybean meal-based diets in a 2 × 4 factorial arrangement, which included 2 CP levels (6.9 or 13.4%) and 4 apparent total tract digestible P (ATTDP) levels (0.09, 0.16, 0.24, or 0.32%). Soybean meal and monocalcium phosphate were used to adjust the CP level and ATTDP level, respectively. Limestone was included in diets to maintain a Ca:ATTDP ratio of 2.5 across treatments. Ileal digesta was continuously collected for 10 h on the last 2 d of each 7-d experimental period. The ileal digesta samples were lyophilized and analyzed to calculate ileal digested N and P. Data were analyzed using PROC MIXED of SAS (9.3) and contrasts were used to test the linear and quadratic effects of increasing levels of P within each CP level. In the low-CP group (6.9%), the ileal digested P was 0.71, 1.16, 1.64, and 2.03 g/kg DMI for diets that contained 0.09, 0.16, 0.24, and 0.32% ATTDP, respectively, and was 0.70, 1.54, 2.03, and 1.99 g/kg DMI, respectively, for their counterparts in the high-CP group (13.4%). A low dietary CP level decreased ( < 0.05) ileal digested P (g/kg DMI). The ileal digested P (g/kg DMI) linearly increased ( < 0.01) with increasing ATTDP levels in the low-CP group, but the pattern was linear ( < 0.01) and quadratic ( < 0.01) in the high-CP group. In the low- and high-CP diets, the determined true ileal digestibility of P in monocalcium phosphate was 54.4 and 75.6%, respectively. In conclusion, this research indicated that the ileal digestion of P could be limited by protein deficiency. The dietary CP level should be considered in P digestibility-related studies.


Participation of follicle associated epithelium (fae), macrophages, and plasma cells in the function of appendix.
Rabbit appendix was studied by correlated scanning and transmission electron microscopy and light microscopy, taking advantage of luminal bacteria as naturally occurring markers for following antigen uptake. The three-dimensional relationship of lymphoid follicles to other structures was revealed clearly. The smooth luminal surface of the appendix is interrupted by stomata each leading to a recessed interval between the dome epithelium covering the apical pole of each follicle and the goblet cell-rich epithelium forming a cupola on the opposing surface. Bacteria abound in this recess. They are taken up by follicle associated epithelial (FAE) cells which comprise part of the dome epithelium. Intraepithelial macrophages phagocytize the bacteria. Bacteria-laden macrophages are numerous in the dome area of the follicle and in the germinal center. Large lymphatic sinuses surrounding the follicles are well visualized following stimulation of fluid movement. Plasma cells are located within the dome epithelial layer and immediately beneath it. Plasma cells may be concentrated along the base of epithelial cells, with the absence of a basal lamina indicating an absence of the molecular selection which it would normally provide. The appendix, like Peyer's patches, takes up antigens, processes them, and gives evidence of reaction to them locally, with the secretions from plasma cells possibly modulating antigen entry.


Identification and characterization of a novel β-D-galactosidase that releases pyruvylated galactose.
Pyruvyl modification of oligosaccharides is widely seen in both prokaryotes and eukaryotes. Although the biosynthetic mechanisms of pyruvylation have been investigated, enzymes that metabolize and degrade pyruvylated oligosaccharides are not well known. Here, we searched for a pyruvylated galactose (PvGal)-releasing enzyme by screening soil samples. We identified a Bacillus strain, as confirmed by the 16S ribosomal RNA gene analysis, that exhibited PvGal-ase activity toward p-nitrophenyl-β-D-pyruvylated galactopyranose (pNP-β-D-PvGal). Draft genome sequencing of this strain, named HMA207, identified three candidate genes encoding potential PvGal-ases, among which only the recombinant protein encoded by ORF1119 exhibited PvGal-ase activity. Although ORF1119 protein displayed broad substrate specificity for pNP sugars, pNP-β-D-PvGal was the most favorable substrate. The optimum pH for the ORF1119 PvGal-ase was determined as 7.5. A BLAST search suggested that ORF1119 homologs exist widely in bacteria. Among two homologs tested, BglC from Clostridium but not BglH from Bacillus showed PvGal-ase activity. Crystal structural analysis together with point mutation analysis revealed crucial amino acids for PvGal-ase activity. Moreover, ORF1119 protein catalyzed the hydrolysis of PvGal from galactomannan of Schizosaccharomyces pombe, suggesting that natural polysaccharides might be substrates of the PvGal-ase. This novel PvGal-catalyzing enzyme might be useful for glycoengineering projects to produce new oligosaccharide structures.


New earth system model for optical performance evaluation of space instruments.
In this study, a new global earth system model is introduced for evaluating the optical performance of space instruments. Simultaneous imaging and spectroscopic results are provided using this global earth system model with fully resolved spatial, spectral, and temporal coverage of sub-models of the Earth. The sun sub-model is a Lambertian scattering sphere with a 6-h scale and 295 lines of solar spectral irradiance. The atmospheric sub-model has a 15-layer three-dimensional (3D) ellipsoid structure. The land sub-model uses spectral bidirectional reflectance distribution functions (BRDF) defined by a semi-empirical parametric kernel model. The ocean is modeled with the ocean spectral albedo after subtracting the total integrated scattering of the sun-glint scatter model. A hypothetical two-mirror Cassegrain telescope with a 300-mm-diameter aperture and 21.504 mm × 21.504-mm focal plane imaging instrument is designed. The simulated image results are compared with observational data from HRI-VIS measurements during the EPOXI mission for approximately 24 h from UTC Mar. 18, 2008. Next, the defocus mapping result and edge spread function (ESF) measuring result show that the distance between the primary and secondary mirror increases by 55.498 μm from the diffraction-limited condition. The shift of the focal plane is determined to be 5.813 mm shorter than that of the defocused focal plane, and this result is confirmed through the estimation of point spread function (PSF) measurements. This study shows that the earth system model combined with an instrument model is a powerful tool that can greatly help the development phase of instrument missions.


Unusual clot formation following insertion of central double lumen catheter for hemodialysis.
We report a case of a patient with chronic renal failure (CRF) due to fibrillary glomerulonephritis, who was admitted to our Hospital with fever accompanied by chills, of two days duration. Due to aggravation of his CRF, accompanied by severe metabolic acidosis, an internal jugular double lumen central vein catheter was inserted, through which he was hemodialyzed. Blood cultures were negative. The catheter was removed and the tip was cultured; it revealed colonization with hemolytic Staphylococcus sensitive to vancomycin and metilmycin. They were both given to the patient with excellent results. Trans-thoracic and trans-esophageal echocardiography revealed a clot in the right atrium with an abnormal circumference, connected to the tip of the second catheter. Anticoagulants were administered (calsium nadroparin, converted seven days later to acenocoumarol) and the size of the clot was significantly reduced. The case is presented in order to prove the significant contribution of trans-esophageal echocardiography as a non-invasive imaging technique for the detection of a clot in the right atrium and the impressive influence of appropriate therapeutic management in saving the patient's life and reducing the clot size. The trans-esophageal echocardiogram provides invaluable information concerning the evolution, development and regression of right atrium clots formed at the tip of jugular catheters.


Spectroscopic insights into the decreased efficiency of chlorosomes containing bacteriochlorophyll f.
Chlorosomes are light-harvesting antenna complexes that occur in green photosynthetic bacteria which have only been shown naturally to contain bacteriochlorophyll (BChl) c, d, or e as the principal light-harvesting pigments. BChl f has long been thought to be an obvious fourth member of the so-called Chlorobium chlorophylls, because it possesses a C-7 formyl group like BChl e and lacks a methyl group at C-20 like BChl d. In organisms that synthesize BChl c or e, the bchU gene product encodes the enzyme that methylates the C-20 position of these molecules. A bchU null mutant of the green sulfur bacterium Chlorobaculum limnaeum strain 1677(T), which normally synthesizes BChl e, has recently been generated via insertional inactivation, and it produces chlorosomes containing BChl f [Vogl et al., 2012]. In this study, chlorosomes containing BChl f and monomeric BChl f in pyridine were characterized using a variety of spectroscopic techniques, including fluorescence emission and excitation spectroscopy, fluorescence lifetime and quantum yield determinations, and circular dichroism. These spectroscopic measurements, as well as Gaussian simulation of the data, show that chlorosomes containing BChl f are less efficient in energy transfer than those with BChl e. This can primarily be attributed to the decreased spectral overlap between the oligomeric BChl f (energy donor) fluorescence emission and the BChl a (energy acceptor) absorption in the chlorosome baseplate. This study allows us to hypothesize that, if they exist in nature, BChl f-containing organisms most likely live in rare high-light, anoxic conditions devoid of Chl a, d, or BChl e filtering. K. Vogl, M. Tank, G.S. Orf, R.E. Blankenship, D.A. Bryant, Bacteriochlorophyll f: properties of chlorosomes containing the "forbidden chlorophyll," Front. Microbiol. 3 (2012) 298.


Pupillary dilation as a measure of sexual arousal. A reply to Hamel.
A comment is presented on a report dealing with pupillary dilation as a measure of sexual interest. Objections are made concerning the use of an earlier paper by the author as a "straw man," and caution pertaining to traditional methodology is offered.


Membrane Functionalization Approaches toward Per- and Polyfluoroalkyl Substances and Selected Metal Ion Separations.
Adsorption and ion exchange technologies are two of the most widely used approaches to separate pollutants from water; however, their intrinsic diffusion limitations continue to be a challenge. Pore functionalized membranes are a promising technology that can help overcome these challenges, but the extents of their competitive benefits and broad applicability have not been systematically evaluated. Herein, three types of adsorptive/ion exchange (IX) polymers containing strong/weak acid, strong base, and iron-chitosan complex groups were synthesized in the pores and partially on the surface of microfiltration (MF) membranes and tested for the removal of organic and inorganic cations and anions from water, including arsenic, per- and polyfluoroalkyl substances (PFAS), and calcium (hardness). When directly compared with beads (0.5-6 mm) and crushed resins (0.05 mm), adsorptive/IX pore-functionalized membranes demonstrated an increased relative sorption capacity, up to 2 orders of magnitude faster kinetics and the ability to regenerate up to 70-100% of their capacity while concentrating the initial solution concentration up to 12 times. The simple and versatile synthesis approach used to functionalize membranes, notably independent of the polymer type of the MF membrane, utilized pores throughout the entire cross section of the membrane to immobilize the polymers that contain the functional groups. Utilizing the pore volume of commercial membranes (6-112 mL/m2), the scientific weight capacity of the polymer (3.1-11.5 mequiv/g), and the synthesis conditions (e.g., monomer concentration), the theoretical adsorption/IX capacities per area of the membranes were calculated to be as high as 550 mequiv/m2, substantially higher than the 175 mequiv/m2 value needed to compete with commercially available IX resins. This work therefore shows that pore functionalized membranes are a promising path to tackle water contamination challenges, lowering separation diffusion limitations.


Research and development of Zika virus vaccines.
Zika virus (ZIKV) is a member of the family Flaviviridae, genus Flavivirus, and is transmitted by Aedes sp. mosquitoes. There are three genetic lineages of ZIKV: the East African, West African and Asian lineages. Until recently, Zika fever (ZF) has normally been considered a rare, mild febrile disease, but reports since 2012 have shown potentially severe complications associated with ZIKV infection, including microcephaly and Guillain-Barré syndrome. There are no licensed vaccines for ZIKV; however, many vaccine platforms/approaches that have been utilised for other flavivirus vaccines are being applied to ZIKV. Given the current outbreak of ZIKV in the Americas with its associated risks to pregnancy, we summarise what is known about the virus, how knowledge of currently licensed flavivirus vaccines can be applied to ZIKV vaccine development and the assessments of potential challenges for ZIKV vaccine testing and evaluation.


Effects of bacterial lipopolysaccharide on the induction and expression of cell-mediated immunity. I. Depression of the afferent arc.
Bacterial lipopolysaccharide (LPS), being a B cell mitogen, enhances antibody production. When given prior to an injection of sheep red blood cells (SRBC) it caused a premature but not significantly increased production of antibody forming cells in regionally stimulated lymph nodes. There was a corresponding depression of T cell activity, as measured by the relative levels of delayed-type hypersensitivity (DTH) in normal and LPS-treated mice. Although LPS also induced hyperplasia of the reticuloendothelial system, and a consequent reduction in the effective antigenic dose of SRBC, this was not the reason for the reduced level of T cell activity. The opposing effects of LPS on T and B cell responses was due to premature inhibition of the T cell response by an accelerated production of antibody. This caused an earlier onset of the inhibitory mechanism which regulates the T cell response and is specific for the inducing antigen.


Training Paradigms in Hepato-Pancreatico-Biliary Surgery: an Overview of the Different Fellowship Pathways.
Hepato-pancreatico-biliary (HPB) surgery, and the training of HPB surgeons, has evolved significantly over the last several decades. The current state of training in HPB surgery in North America is defined through three main pathways: the Complex General Surgical Oncology (CGSO) ACGME fellowship, the American Society of Transplant Surgeons (ASTS) fellowship, and the Americas Hepatopancreaticobiliary Association (AHPBA) fellowship. These fellowships offer variable experiences in pancreas, liver, and biliary cases, and each pathway offers a unique perspective on HPB surgery. The CGSO ACGME, ASTS, and AHPBA fellowships represent decades of work by the three major surgical leadership stakeholders to improve and ensure quality training of future HPB surgeons. The best care is provided by the HPB surgeon who has been trained to understand the importance of all available treatment options within the context of a multidisciplinary setting. The three fellowship pathways are outlined in this paper with the nuances and variations characteristic of the different training programs highlighted.


A phorbol ester-sensitive kinase catalyzes the phosphorylation of P0 glycoprotein in myelin.
The proposed structural protein of peripheral nerve myelin, P0, has been shown to have several covalent modifications. In addition to being glycosylated, sulfated, and acylated, P0 is phosphorylated, with the intracellular site of this latter addition being in question. By employing nerve injury models that exhibit different levels of P0 biosynthesis in the absence and presence of myelin assembly, we have examined the cellular location of P0 phosphorylation. It is demonstrated that there is comparable P0 phosphorylation in both normal and crush-injured adult rat sciatic nerves, although the level of biosynthesis of P0 differs between these myelin maintaining and actively myelinating nerve models, respectively. The glycoprotein does not appear to be phosphorylated readily in the transected adult sciatic nerve, a preparation in which P0 biosynthesis is observed but that lacks myelin membrane. These observations suggest that the modification is not associated with the biosynthesis or maturation of P0 in the endoplasmic reticulum or Golgi, but that it instead occurs after myelin assembly. That P0 phosphorylation occurs in the normal nerve even when translation is inhibited by cycloheximide treatment lends further support to this conclusion. P0 is shown to be phosphorylated on one or more serine residues, with all or most of the phosphate group(s) being labile as evidenced by pulse-chase analysis. Addition of a biologically active phorbol ester, 12-O-tetradecanoylphorbol-13-acetate or 4 beta-phorbol 12,13-dibutyrate, substantially increases the extent of [32P]orthophosphate incorporation into the glycoprotein of normal and crushed nerve but not transected nerve. Biologically inactive 4 alpha-phorbol 12,13-didecanoate has no effect on P0 phosphorylation. Similarly, the addition of the cyclic AMP analog 8-bromo-cyclic AMP causes no appreciable changes in P0 labeling. These findings indicate that the phorbol ester-sensitive enzyme, protein kinase C, may be responsible for the phosphorylation of P0 within the myelin membrane.


Aging and its effects on inflammation in skeletal muscle at rest and following exercise-induced muscle injury.
The world's elderly population is expanding rapidly, and we are now faced with the significant challenge of maintaining or improving physical activity, independence, and quality of life in the elderly. Counteracting the progressive loss of muscle mass that occurs in the elderly, known as sarcopenia, represents a major hurdle in achieving these goals. Indirect evidence for a role of inflammation in sarcopenia is that markers of systemic inflammation correlate with the loss of muscle mass and strength in the elderly. More direct evidence is that compared with skeletal muscle of young people, the number of macrophages is lower, the gene expression of several cytokines is higher, and stress signaling proteins are activated in skeletal muscle of elderly people at rest. Sarcopenia may also result from inadequate repair and chronic maladaptation following muscle injury in the elderly. Macrophage infiltration and the gene expression of certain cytokines are reduced in skeletal muscle of elderly people compared with young people following exercise-induced muscle injury. Further research is required to identify the cause(s) of inflammation in skeletal muscle of elderly people. Additional work is also needed to expand our understanding of the cells, proteins, and transcription factors that regulate inflammation in the skeletal muscle of elderly people at rest and after exercise. This knowledge is critical for devising strategies to restrict sarcopenia, and improve the health of today's elderly population.


Beyond Object Proposals: Random Crop Pooling for Multi-Label Image Recognition.
Learning high-level image representations using object proposals has achieved remarkable success in multi-label image recognition. However, most object proposals provide merely coarse information about the objects, and only carefully selected proposals can be helpful for boosting the performance of multi-label image recognition. In this paper, we propose an object-proposal-free framework for multi-label image recognition: random crop pooling (RCP). Basically, RCP performs stochastic scaling and cropping over images before feeding them to a standard convolutional neural network, which works quite well with a max-pooling operation for recognizing the complex contents of multi-label images. To better fit the multi-label image recognition task, we further develop a new loss function-the dynamic weighted Euclidean loss-for the training of the deep network. Our RCP approach is amazingly simple yet effective. It can achieve significantly better image recognition performance than the approaches using object proposals. Moreover, our adapted network can be easily trained in an end-to-end manner. Extensive experiments are conducted on two representative multi-label image recognition data sets (i.e., PASCAL VOC 2007 and PASCAL VOC 2012), and the results clearly demonstrate the superiority of our approach.


Toward a single-layer two-dimensional honeycomb supramolecular organic framework in water.
The self-assembly of well-defined 2D supramolecular polymers in solution has been a challenge in supramolecular chemistry. We have designed and synthesized a rigid stacking-forbidden 1,3,5-triphenylbenzene compound that bears three 4,4'-bipyridin-1-ium (BP) units on the peripheral benzene rings. Three hydrophilic bis(2-hydroxyethyl)carbamoyl groups are introduced to the central benzene ring to suppress 1D stacking of the triangular backbone and to ensure solubility in water. Mixing the triangular preorganized molecule with cucurbit[8]uril (CB[8]) in a 2:3 molar ratio in water leads to the formation of the first solution-phase single-layer 2D supramolecular organic framework, which is stabilized by the strong complexation of CB[8] with two BP units of adjacent molecules. The periodic honeycomb 2D framework has been characterized by various (1)H NMR spectroscopy, dynamic light scattering, X-ray diffraction and scattering, scanning probe and electron microscope techniques and by comparing with the self-assembled structures of the control systems.


Intestinal carbamoyl phosphate synthase I in human and rat. Expression during development shows species differences and mosaic expression in duodenum of both species.
The clinical importance of carbamoyl phosphate synthase I (CPSI) relates to its capacity to metabolize ammonia, because CPSI deficiencies cause lethal serum ammonia levels. Although some metabolic parameters concerning liver and intestinal CPSI have been reported, the extent to which enterocytes contribute to ammonia conversion remains unclear without a detailed description of its developmental and spatial expression patterns. Therefore, we determined the patterns of enterocytic CPSI mRNA and protein expression in human and rat intestine during embryonic and postnatal development, using in situ hybridization and immunohistochemistry. CPSI protein appeared during human embryogenesis in liver at 31-35 e. d. (embryonic days) before intestine (59 e.d.), whereas in rat CPSI detection in intestine (at 16 e.d.) preceded liver (20 e.d.). During all stages of development there was a good correlation between the expression of CPSI protein and mRNA in the intestinal epithelium. Strikingly, duodenal enterocytes in both species exhibited mosaic CPSI protein expression despite uniform CPSI mRNA expression in the epithelium and the presence of functional mitochondria in all epithelial cells. Unlike rat, CPSI in human embryos was expressed in liver before intestine. Although CPSI was primarily regulated at the transcriptional level, CPSI protein appeared mosaic in the duodenum of both species, possibly due to post-transcriptional regulation.


Chaos in learning a simple two-person game.
We investigate the problem of learning to play the game of rock-paper-scissors. Each player attempts to improve her/his average score by adjusting the frequency of the three possible responses, using reinforcement learning. For the zero sum game the learning process displays Hamiltonian chaos. Thus, the learning trajectory can be simple or complex, depending on initial conditions. We also investigate the non-zero sum case and show that it can give rise to chaotic transients. This is, to our knowledge, the first demonstration of Hamiltonian chaos in learning a basic two-person game, extending earlier findings of chaotic attractors in dissipative systems. As we argue here, chaos provides an important self-consistency condition for determining when players will learn to behave as though they were fully rational. That chaos can occur in learning a simple game indicates one should use caution in assuming real people will learn to play a game according to a Nash equilibrium strategy.


Ovatodiolide induces autophagy-mediated cell death through the p62-Keap1-Nrf2 signaling pathway in chronic myeloid leukemia cells.
Ovatodiolide is a macrocyclic diterpenoid compound with various biological activities that displays considerable anticancer potential in different tumor models. However, the underlying mechanism for this antineoplastic activity remains unclear. The aim of the present study was to investigate the anticancer effect and possible molecular mechanism of ovatodiolide in human chronic myeloid leukemia (CML). Ovatodiolide suppressed cell colony formation and induced apoptosis in the K562 and KU812 cells. We also observed that ovatodiolide enhanced the production of reactive oxygen species (ROS), activated Nrf2 signaling, and inhibited mTOR phosphorylation. Autophagic flux was shown to be enhanced after treatment with ovatodiolide in K562 cells. Furthermore, autophagy inhibition alleviated ovatodiolide-induced cell apoptosis, whereas autophagy promotion aggravated apoptosis in CML cells. These results demonstrated that ovatodiolide activates autophagy-mediated cell death in CML cells. Additionally, ovatodiolide transcriptionally activated the expression of p62, and the p62 levels were negatively regulated by autophagy. Moreover, p62-Keap1-Nrf2 signaling was confirmed to be involved in ovatodiolide-induced cell death. Accordingly, LC3B knockdown augmented the ovatodiolide-induced p62 expression, increased the p62-Keap1 interaction, and enhanced the translocation of Nrf2 into the nucleus. In contrast, p62 inhibition abolished the effects that were induced through ovatodiolide treatment. Nrf2 inhibition with ML385 diminished the protective effect of autophagy inhibition in CML cells. Collectively, our results indicate that ovatodiolide induces oxidative stress and provokes autophagy, which effectively decreases the expression of p62 and weakens the protective effect of Nrf2 signaling activation, thus contributing to apoptosis in CML cells.


Development of a novel AIDS vaccine: the HIV-1 transactivator of transcription protein vaccine.
Classical approaches aimed at targeting the HIV-1 envelope as well as other structural viral proteins have largely failed. The HIV-1 transactivator of transcription (Tat) is a key HIV virulence factor, which plays pivotal roles in virus gene expression, replication, transmission and disease progression. Notably, anti-Tat Abs are uncommon in natural infection and, when present, correlate with the asymptomatic state and lead to lower or no disease progression. Hence, targeting Tat represents a pathogenesis-driven intervention. Here, we review the rationale and the translational development of a therapeutic vaccine targeting the Tat protein. Preclinical and Phase I studies, Phase II trials with Tat in anti-Tat Ab-negative, virologically suppressed highly active antiretroviral therapy-treated subjects in Italy and South Africa were conducted. The results indicate that Tat-induced immune responses are necessary to restore immune homeostasis, to block the replenishment and to reduce the size of the viral reservoir. Additionally, they may help in establishing key parameters for highly active antiretroviral therapy intensification and a functional cure. We propose the therapeutic setting as the most feasible to speed up the testing and comparison of preventative vaccine candidates, as the distinction lies in the use of the vaccine in uninfected versus infected subjects and not in the vaccine formulation.


Sports activity participation after curved periacetabular osteotomy for acetabular dysplasia.
Curved periacetabular osteotomy (CPO) was developed to treat acetabular dysplasia. Given that CPO can improve physical function in the early post-operative period, patients might be able to participate in sports activities post-operatively. Therefore, this study examined the post-operative sports activity participation and characteristics of acetabular dysplasia patients who have undergone CPO. A total of 52 patients who underwent CPO for acetabular dysplasia were given a questionnaire on pre- and post-operative sports activities; 43 patients responded. We surveyed patients' sports activities, satisfaction, and physical function. Patients were divided according to whether they participated in sports activities after CPO. Physical function was compared before and after CPO. The pre- and post-operative sports activity participation rates were 55.8 and 72.1%, respectively. Patients mostly performed low-impact sports activities. Moreover, patients who participated in sports activities post-operatively had smaller pre-operative range of motion of hip flexion and returned to full weight bearing earlier. Among acetabular dysplasia patients who underwent CPO, 72.1% participated in sports activities post-operatively. Post-operatively, patients participated not only in low-impact sports activities, but also in high-impact ones. These findings might be useful for advising patients who are concerned about participating in sports activities after CPO.


Self-Powered Perovskite/CdS Heterostructure Photodetectors.
Methylammonium lead halide perovskites have gained a lot of attention because of their remarkable physical properties and potential for numerous (opto)electronic applications. Here, high-performance photodetectors based on CH3NH3PbI3 (MAPbI3)/CdS heterostructures are demonstrated. The resulting self-powered MAPbI3/CdS photodetectors show excellent operating characteristics including a maximum detectivity of 2.3 × 1011 Jones with a responsivity of 0.43 A/W measured at 730 nm. A temporal response time of less than 14 ms was achieved. The mechanisms of charge separation and transport at the interface of the MAPbI3/CdS junction were investigated via conductive atomic force microscopy (AFM) and photoconductive AFM. Obtained results show that grain boundaries exhibit higher photocurrent than flat regions of the top perovskite layer, which indicates that excitons preferentially separate at the grain boundaries of the perovskite thin film, that is, at the edges of the MAPbI3 crystals. The study of the photoelectric mechanism at the nanoscale suggests the device performance could potentially be fine-tuned through grain boundary engineering, which provides essential insights for the fabrication of the high-performance photodetector. The demonstrated self-powered photodetector is promising for numerous applications in low-energy consumption optoelectronic devices.


Prostacyclin synthase expression and epigenetic regulation in nonsmall cell lung cancer.
Prostacyclin synthase (PGIS) metabolizes prostaglandin H(2), into prostacyclin. This study aimed to determine the expression profile of PGIS in nonsmall cell lung cancer (NSCLC) and examine potential mechanisms involved in PGIS regulation. PGIS expression was examined in human NSCLC and matched controls by reverse transcriptase polymerase chain reaction (RT-PCR), Western analysis, and immunohistochemistry. A 204-patient NSCLC tissue microarray was stained for PGIS and cyclooxygenase 2 (COX2) expression. Staining intensity was correlated with clinical parameters. Epigenetic mechanisms underpinning PGIS promoter expression were examined using RT-PCR, methylation-specific PCR, and chromatin immunoprecipitation analysis. PGIS expression was reduced/absent in human NSCLC protein samples (P < .0001), but not mRNA relative to matched controls. PGIS tissue expression was higher in squamous cell carcinoma (P = .004) and in male patients (P < .05). No significant correlation of PGIS or COX2 expression with overall patient survival was observed, although COX2 was prognostic for short-term (2-year) survival (P < .001). PGIS mRNA expression was regulated by DNA CpG methylation and histone acetylation in NSCLC cell lines, with chromatin remodeling taking place directly at the PGIS gene. PGIS mRNA expression was increased by both demethylation agents and histone deacetylase inhibitors. Protein levels were unaffected by demethylation agents, whereas PGIS protein stability was negatively affected by histone deacetylase inhibitors. PGIS protein expression is reduced in NSCLC, and does not correlate with overall patient survival. PGIS expression is regulated through epigenetic mechanisms. Differences in expression patterns between mRNA and protein levels suggest that PGIS expression and protein stability are regulated post-translationally. PGIS protein stability may have an important therapeutic role in NSCLC.


Economic analysis of an internet-based depression prevention intervention.
The transition through adolescence places adolescents at increased risk of depression, yet care-seeking in this population is low, and treatment is often ineffective. In response, we developed an Internet-based depression prevention intervention (CATCH-IT) targeting at-risk adolescents. We explore CATCH-IT program costs, especially safety costs, in the context of an Accountable Care Organization as well as the perceived value of the Internet program. Total and per-patient costs of development were calculated using an assumed cohort of a 5,000-patient Accountable Care Organization. Total and per-patient costs of implementation were calculated from grant data and the Medicare Resource-Based Relative Value Scale (RBRVS) and were compared to the willingness-to-pay for CATCH-IT and to the cost of current treatment options. The cost effectiveness of the safety protocol was assessed using the number of safety calls placed and the percentage of patients receiving at least one safety call. The willingness-to-pay for CATCH-IT, a measure of its perceived value, was assessed using post-study questionnaires and was compared to the development cost for a break-even point. We found the total cost of developing the intervention to be USD 138,683.03. Of the total, 54% was devoted to content development with per patient cost of USD 27.74. The total cost of implementation was found to be USD 49,592.25, with per patient cost of USD 597.50. Safety costs accounted for 35% of the total cost of implementation. For comparison, the cost of a 15-session group cognitive behavioral therapy (CBT) intervention aimed at at-risk adolescents was USD 1,632 per patient. Safety calls were successfully placed to 96.4% of the study participants. The cost per call was USD 40.51 with a cost per participant of USD 197.99. The willingness-to-pay for the Internet portion of CATCH-IT had a median of USD 40. The break-even point to offset the cost of development was 3,468 individuals. Developing Internet-based interventions like CATCH-IT appears economically viable in the context of an Accountable Care Organization. Furthermore, while the cost of implementing an effective safety protocol is proportionally high for this intervention, CATCH-IT is still significantly cheaper to implement than current treatment options. Limitations of this research included diminished participation in follow-up surveys assessing willingness-to-pay. IMPLICATIONS FOR HEALTH CARE PROVISION AND USE AND HEALTH POLICIES: This research emphasizes that preventive interventions have the potential to be cheaper to implement than treatment protocols, even before taking into account lost productivity due to illness. Research such as this business application analysis of the CATCH-IT program highlights the importance of supporting preventive medical interventions as the healthcare system already does for treatment interventions. This research is the first to analyze the economic costs of an Internet-based intervention. Further research into the costs and outcomes of such interventions is certainly warranted before they are widely adopted. Furthermore, more research regarding the safety of Internet-based programs will likely need to be conducted before they are broadly accepted.


High-resolution study of spin excitations in the singlet ground state of SrCu2(BO3)2.
High-resolution, inelastic neutron scattering measurements on SrCu2(BO3)2, a realization of the Shastry-Sutherland model for two-dimensional Heisenberg antiferromagnets, reveal the dispersion of the three single triplet excitations continuously across the (H,0) direction within its tetragonal basal plane. These measurements also show distinct Q dependencies for the single and multiple triplet excitations, and that these excitations are largely dispersionless perpendicular to this plane. The temperature dependence of the intensities of these excitations is well described as the complement of the dc susceptibility of SrCu2(BO3)2.


Dodine as a Kosmo-Chaotropic Agent.
Denaturants such as the guanidinium cation unfold proteins at molar concentrations, which interferes with ultraviolet- and infrared-based spectroscopy measurements. Dodine denatures some proteins cooperatively at a thousand-fold lower concentration, allowing for spectroscopy measurements. Nonetheless, dodine's microscopic mechanism of interaction with proteins is not understood. We probe the effect of dodine on α-helices and tertiary structure by investigating the stability of the small helical protein B. Experiments show that dodine promotes formation of helical structure (a kosmotropic effect), while inducing the loss of tertiary structure (a chaotropic effect). Although dodine destabilizes native protein structure, it does not lower the thermal denaturation midpoint temperature of protein B. All-atom simulations reveal the cause for both observations: The denaturant action of dodine's guanidyl headgroup is counteracted by its aliphatic tail, which stabilizes amphipathic helices and associates with an expanded protein core. The Janus-like behavior of headgroup and tail make dodine a simultaneous stabilizer-destabilizer or "kosmo-chaotrope".


Eye-lens accommodation load and static trapezius muscle activity.
The purpose of this experimental study was to investigate if sustained periods of oculomotor load impacts on neck/scapular area muscle activity. The static trapezius muscle activity was assessed from bipolar surface electromyography, normalized to a submaximal contraction. Twenty-eight subjects with a mean age of 29 (range 19-42, SD 8) viewed a high-contrast fixation target for two 5-min periods through: (1) -3.5 dioptre (D) lenses; and (2) 0 D lenses. The target was placed 5 D away from the individual's near point of accommodation. Each subject's ability to compensate for the added blur was extracted via infrared photorefraction measurements. Subjects whose accommodative response was higher in the -D blur condition (1) showed relatively more static bilateral trapezius muscle activity level. During no blur (2) there were no signs of relationships. The results indicate that sustained eye-lens accommodation at near, during ergonomically unfavourable viewing conditions, could possibly represent a risk factor for trapezius muscle myalgia.


Estimation of the orientation term of the general quadrature transform from a single n-dimensional fringe pattern.
The spatial orientation of the fringe has been demonstrated to be a key point in the reliable phase demodulation from a single n-dimensional fringe pattern regardless of the frequency spectrum of the signal. The recently introduced general n-dimensional quadrature transform (GQT) makes explicit the importance of the fringe orientation in the demodulation process. The GQT is a quadrature operator that transforms cos phi into -sin phi--where phi is the modulating phase--and it is composed of two terms: an orientation factor directly related to the fringe's spatial orientation and an isotropic n-dimensional generalization of the one-dimensional Hilbert transform. We present a method for the determination of the orientation factor in a general n-dimensional case and its application to the demodulation of a single fringe pattern by the GQT. We have tested the algorithm with simulated as well as real photoelastic fringe patterns with good results.


Using fuzzy logic approach to predict work-related musculoskeletal disorders among automotive assembly workers.
Background: Musculoskeletal disorders (MSDs) are considered an important health concern, particularly in automotive assembly industries. Evaluation of the effects of all MSDs risk factors is difficult due to its multifactorial nature. In addition, the risk factors cannot be detected accurately when they are only based on individual opinions. Thus, in this study, fuzzy logic tool was used to evaluate the combined effects of all risk factors on MSDs. Methods: This cross sectional study was conducted on 100 male workers in an automotive industry. Job satisfaction, job stress, job fatigue, and body posture were evaluated by a self-reported questionnaire. Body posture was evaluated using Rapid Entire Body Assessment (REBA). Primary data analysis on extracting the input variables of MATLAB was performed by SPSS 22, with a significant level of 0.05. T test, one-way Anova, and Pearson correlation analysis were used to extract the input variables for the fuzzy logic model. The results obtained from the Nordic questionnaire was selected as the output of the fuzzy model. Fuzzy logic assessment was performed using MATLAB software version 7.0. Results: There were significant differences between WMSDs factors, including job fatigue, strain, working posture, and the REBA final score, and pain in all limbs of the body (p<0.05). A significant difference was also found between working posture with wrist score (p<0.05). The findings on defuzzification showed a strong correlation between real and modelling results. Conclusion: The results showed that many factors such as posture, fatigue, and strain affect MSDs. Based on the obtained results, all categories of risk factors, including personal, psychosocial, and occupational, should be considered to predict MSDs, which can be achieved by a modeling approach.


Spatiotemporal variation of dissolved oxygen concentration in the northern Gulf of Mexico during the period of 1992-2017.
Dissolved oxygen (DO) concentration is an important index of ocean systems. In this study, spatiotemporal variations in DO were analyzed in the northern Gulf of Mexico (NGM) for the period of 1992-2017. Temporal variation involves annual, monthly, and seasonal variations. The spatial scope ranged from the sea surface to 1000 m depth. The results show that: (1) DO decreased from 1992 to 2017 with a brief increase; (2) clear seasonal variation characteristics of DO were presented, and the seasonal average DO was relatively low in summer; (3) the monthly average trends of sea surface DO in different years were relatively consistent; and (4) with increasing water depth from seawater surface to 1000 m, the average DO of 1992 to 2017 tended to decrease and then increase. The results aim at shedding some light on the assessment of DO change in the NGM over the past quarter century.


PPARs in Neurodegenerative and Neuroinflammatory Pathways.
PPARs are lipid sensors activated by dietary lipids or their metabolites, mainly fatty acids and eicosanoids, that play critical roles in CNS biology, since brain has a very high lipid content and has the higher energetic metabolism in the body. In neurodegenerative diseases in addition to metabolic impairment, also neuroinflammation is observed and PPARs are also closely linked to inflammatory processes. Several studies have revealed a complicated relationship between the innate immune response and tissue metabolism. In the brain, during pathological conditions, an alteration in metabolic status occurs, particularly involving glucose utilization and production, a condition which is generally related to metabolic changes. Taking into account the high expression of PPARs in the brain, this review will focus on the role of these transcription factors in CNS diseases.


Hemp (Cannabis sativa L.) Seed Phenylpropionamides Composition and Effects on Memory Dysfunction and Biomarkers of Neuroinflammation Induced by Lipopolysaccharide in Mice.
Hempseed has achieved a growing popularity in human nutrition, particularly regarding essential amino acids and fatty acids. The multiple positive attributes of hempseed have led to the further study of its constituents. In this study, hempseed extract containing phenylpropionamides (TPA) was obtained and its chemical profile and content were obtained using high-performance liquid chromatography technology based on previous study. The anti-neuroinflammatory effect of TPA extract was evaluated using a lipopolysaccharide (LPS)-induced mouse model. Fourteen phenylpropionamides (TPA) were identified in the obtained extract with a total content of 233.52 ± 2.50 μg/mg extract. In mice, TPA prevented the learning and spatial memory damage induced by LPS. Increased brain levels of IL-1β, IL-6, and TNF-α in the LPS-induced mice were reduced by TPA treatment. Furthermore, TPA attenuated LPS-induced hippocampal neuronal damage in mice. This study demonstrates the nutraceutical potential of hempseed from a neuroprotective perspective.


Comparative chromosome painting in hummingbirds (Trochilidae).
Hummingbirds (Trochilidae) are one of the most enigmatic avian groups, and also among the most diverse, with approximately 360 recognized species in 106 genera, of which 43 are monotypic. This fact has generated considerable interest in the evolutionary biology of the hummingbirds, which is reflected in a number of DNA-based studies. However, only a few of them explored chromosomal data. Given this, the present study provides an analysis of the karyotypes of three species of Neotropical hummingbirds, Anthracothorax nigricollis (ANI), Campylopterus largipennis (CLA), and Hylocharis chrysura (HCH), in order to analyze the chromosomal processes associated with the evolution of the Trochilidae. The diploid number of ANI is 2n=80 chromosomes, while CLA and HCH have identical karyotypes, with 2n=78. Chromosome painting with Gallus gallus probes (GGA1-12) shows that the hummingbirds have a karyotype close to the proposed ancestral bird karyotype. Despite this, an informative rearrangement was detected: an in-tandem fusion between GGA7 and GGA9 found in CLA and HCH, but absent in ANI. A comparative analysis with the tree of life of the hummingbirds indicated that this fusion must have arisen following the divergence of a number of hummingbird species.


Methanogenesis in the sediments of Rio Tinto, an extreme acidic river.
Río Tinto (Iberian Pyritic Belt, SW Spain) is well known for its low pH (mean pH 2.3), high redox potential (> +400 mV) and high concentration of heavy metals. In this work we describe and analyse the presence of methanogenic archaea in the extreme acidic and oxidizing environment of the Tinto basin. Methane formation was measured in microcosms inoculated with sediments from the Rio Tinto basin. Methanol, formate, volatile fatty acids and lactate stimulated the production of methane. Methane formation was associated with a decrease of redox potential and an increase in pH. Cores showed characteristic well-defined black bands in which a high acetate concentration was measured among the otherwise reddish-brown sediments with low acetate concentration. Methanosaeta concilii was detected in the black bands. In enrichment cultures, M. concilii (enriched with a complex substrate mixture), Methanobacterium bryantii (enriched with H(2)) and Methanosarcina barkeri (enriched with methanol) were identified. Our results suggest that methanogens thrive in micro-niches with mildly acidic and reducing conditions within Rio Tinto sediments, which are, in contrast, immersed in an otherwise extremely acidic and oxidizing environment.


PGP 9.5 distribution patterns in biopsies from early lesions of atopic dermatitis.
Peripheral nerve fibres are often increased in lesional skin of atopic dermatitis (AD) patients. We attempted to study nerve fibre profiles, using PGP 9.5 as neuronal marker, in early AD lesions in 10 patients, as compared to non-lesional skin in the same patients and skin from healthy controls. The number of PGP 9.5-positive nerve fibre profiles was not different in the biopsies taken from normal-looking AD skin and healthy controls. The total number of PGP 9.5-positive nerve fibre profiles in the whole skin sections was higher in both the epidermis and the dermis in the group of skin biopsies taken from early lesions of AD patients. Further, the number of epidermal PGP 9.5-positive dendritic cells was increased in AD skin. It seems reasonable that PGP 9.5-positive nerve fibres and PGP 9.5-positive dendritic cells have pathological roles in AD. The findings might serve as a basis for further studies in evaluating novel diagnostic and therapeutic approaches.


AMP-activated protein kinase inhibits transforming growth factor-beta-induced Smad3-dependent transcription and myofibroblast transdifferentiation.
In wound healing, myofibroblast transdifferentiation (MFT) is a metaplastic change in phenotype producing profibrotic effector cells that secrete and remodel the extracellular matrix. Unlike pathways that induce MFT, the molecular mechanisms that negatively regulate MFT are poorly understood. Here, we report that AMP-activated protein kinase (AMPK) blocks MFT in response to transforming growth factor-beta (TGFbeta). Pharmacological activation of AMPK inhibited TGFbeta-induced secretion of extracellular matrix proteins collagen types I and IV and fibronectin. AMPK activation also prevented induction of the myofibroblast phenotype markers alpha-smooth muscle actin and the ED-A fibronectin splice variant. AMPK activators did not prevent MFT in cells transduced with an adenovirus expressing dominant negative, kinase-dead AMPKalpha2. Moreover, AMPK activators did not inhibit MFT induction in AMPK(alpha1,2)(-/-) fibroblasts, demonstrating a requirement for AMPK(alpha) expression. Adenoviral transduction of constitutively active AMPK(alpha2) was sufficient to prevent TGFbeta-induced collagen I, alpha-smooth muscle actin, and ED-A fibronectin. AMPK did not reduce TGFbeta-stimulated Smad3 COOH-terminal phosphorylation and nuclear translocation, which are necessary for MFT. However, AMPK activation inhibited TGFbeta-induced transcription driven by Smad3-binding cis-elements. Consistent with a role for AMPK in transcriptional regulation, nuclear translocation of AMPKalpha2 correlated with the appearance of active AMPKalpha in the nucleus. Collectively, these results demonstrate that AMPK inhibits TGFbeta-induced transcription downstream of Smad3 COOH-terminal phosphorylation and nuclear translocation. Furthermore, activation of AMPK is sufficient to negatively regulate MFT in vitro.


Epigallocatechin-3-gallate inhibits IL-6 synthesis and suppresses transsignaling by enhancing soluble gp130 production.
Regulation of IL-6 transsignaling by the administration of soluble gp130 (sgp130) receptor to capture the IL-6/soluble IL-6R complex has shown promise for the treatment of rheumatoid arthritis (RA). However, enhancing endogenous sgp130 via alternative splicing of the gp130 gene has not yet been tested. We found that epigallocatechin-3-gallate (EGCG), an anti-inflammatory compound found in green tea, inhibits IL-1beta-induced IL-6 production and transsignaling in RA synovial fibroblasts by inducing alternative splicing of gp130 mRNA, resulting in enhanced sgp130 production. Results from in vivo studies using a rat adjuvant-induced arthritis model showed specific inhibition of IL-6 levels in the serum and joints of EGCG-treated rats by 28% and 40%, respectively, with concomitant amelioration of rat adjuvant-induced arthritis. We also observed a marked decrease in membrane-bound gp130 protein expression in the joint homogenates of the EGCG-treated group. In contrast, quantitative RT-PCR showed that the gp130/IL-6Ralpha mRNA ratio increased by approximately 2-fold, suggesting a possible mechanism of sgp130 activation by EGCG. Gelatin zymography results showed EGCG inhibits IL-6/soluble IL-6R-induced matrix metalloproteinase-2 activity in RA synovial fibroblasts and in joint homogenates, possibly via up-regulation of sgp130 synthesis. The results of these studies provide previously undescribed evidence of IL-6 synthesis and transsignaling inhibition by EGCG with a unique mechanism of sgp130 up-regulation, and thus hold promise as a potential therapeutic agent for RA.


Low molecular weight heparin-catalyzed inactivation of factor Xa and thrombin by antithrombin III--effect of platelet factor 4.
Low molecular weight (LMW) heparin preparations have unknown distributions of ATIII-binding material, so mean molecular weights as such might bear little information on their anti-factor Xa and anti-thrombin activities, and on the neutralization of these activities by platelet factor 4 (PF4). These properties were investigated in pure systems with proteins of human origin. Pseudo-first order rate constants of inactivation of factor Xa and thrombin by antithrombin III were determined as function of heparin concentration, in the presence of 4.0 mM CaCl2. Despite a large variation in the mean molecular weights, the ratios of the anti-factor Xa over the anti-thrombin activities were essentially the same for the 4th International Standard for heparin (0.46), the 1st International Standard for LMW heparin (0.32), CY216 (0.42) and enoxaparin (0.50). The ultra LMW heparin CY222 had only a 2-times higher ratio (0.98). Analysis of CY216 subfractions, obtained by gel filtration, showed that the heparin molecules of the upper region of the molecular weight distribution are responsible for the anti-thrombin, but also to a large extent for the anti-factor Xa activities. The results indicate that depolymerization of unfractionated heparin does not result in an increased anti-factor Xa/anti-thrombin ratio, because in the presence of Ca(2+)-ions the rate constants of inactivation of factor Xa are lowered as compared to those of native heparin. PF4-dependent neutralization of anti-factor Xa and anti-thrombin activities of fixed concentrations of the LMW heparins was studied by measuring rate constants as function of PF4 concentration.(ABSTRACT TRUNCATED AT 250 WORDS)


An enzyme with a deep trefoil knot for the active-site architecture.
Knots in polypeptide chains have been found in very few proteins. Only two proteins are considered to have a shallow 'trefoil' knot, which tucks a few residues at one end of the chain through a loop exposed on the protein surface. Recently, another protein was found by a mathematical algorithm to have a deep 'figure-of-eight' knot which had not been visually identified. In the present study, the crystal structure of a hypothetical RNA 2'-O-ribose methyltransferase from Thermus thermophilus (RrmA) was determined at 2.4 A resolution and a deep trefoil knot was found for the first time. The present knot is formed by the threading of a 44-residue polypeptide chain through a 41-residue loop and is better defined than the previously reported knots. Two of the three catalytic residues conserved in the 2'-O-ribose methyltransferase family are located in the knotting loop and in the knotted carboxy-terminal chain, which is the first observation that the enzyme active site is constructed right on the knot. On the other hand, the amino-terminal domain exhibits a geometrical similarity to the ribosomal proteins which recognize an internal loop of RNA.


Cell death by necrosis, a regulated way to go.
Apoptosis is a programmed form of cell death with well-defined morphological traits that are often associated with activation of caspases. More recently evidence has become available demonstrating that upon caspase inhibition alternative programs of cell death are executed, including ones with features characteristic of necrosis. These findings have changed our view of necrosis as a passive and essentially accidental form of cell death to that of an active, regulated and controllable process. Also necrosis has now been observed in parallel with, rather than as an alternative pathway to, apoptosis. Thus, cell death responses are extremely flexible despite being programmed. In this review, some of the hallmarks of different programmed cell death modes have been highlighted before focusing the discussion on necrosis. Obligatory events associated with this form of cell death include uncompensated cell swelling and related changes at the plasma membrane. In this context, representatives of the transient receptor channel family and their regulation are discussed. Also mechanisms that lead to execution of the necrotic cell death program are highlighted. Emphasis is laid on summarizing our understanding of events that permit switching between cell death modes and how they connect to necrosis. Finally, potential implications for the treatment of some disease states are mentioned.


The correlation between the SOS in trabecular bone and stiffness and density studied by finite-element analysis.
For the clinical assessment of osteoporosis (i.e., a degenerative bone disease associated with increased fracture risk), ultrasound has been proposed as an alternative or supplement to the dual-energy X-ray absorptiometry (DEXA) technique. However, the interaction of ultrasound waves with (trabecular) bone remains relatively poorly understood. The present study aimed to improve this understanding by simulating ultrasound wave propagation in 15 trabecular bone samples from the human lumbar spine, using microcomputed tomography-based finite-element modeling. The model included only the solid bone, without the bone marrow. Two structural parameters were calculated: the bone volume fraction (BV/TV) and the structural (apparent) elastic modulus (E(s)), and the ultrasound propagation parameter speed of sound (SOS). Relations between BV/TV and E(s) were similar to published experimental relations. At 1 MHz, correlations between SOS and the structural parameters BV/TV and Es were rather weak, but the results can be explained from the specific features of the trabecular structure and the intrinsic material elastic modulus E(i). In particular, the systematic differences between the three main directions provide information on the trabecular structure. In addition, at 1 MHz the correlation found between the simulated SOS values and those calculated from the simple bar equation was poor when the three directions are considered separately. Hence, under these conditions, the homogenization approach-including the bar equation-is not valid. However, at lower frequencies (50-300 kHz) this correlation significantly improved. It is concluded that detailed analysis of ultrasound wave propagation through the solid structure in various directions and with various frequencies, can yield much information on the structural and mechanical properties of trabecular bone.


Multimodal signaling in fowl, Gallus gallus.
Many social birds produce food-associated calls. In galliforms, these vocalizations are typically accompanied by a distinctive visual display, creating a multimodal signal known as tidbitting. This system is ideal for experimental analysis of the way in which signal components interact to determine overall efficacy. We used high-definition video playback to explore perception of male tidbitting by female fowl, Gallus gallus. Hens experienced four treatments consisting of multimodal tidbitting, visual tidbitting without sound, audible tidbitting without a male present, and a silent empty cage control. Hens took longer to begin food search when the display was silent, but the overall rate of this response did not differ among the multimodal, visual only or audio only playback treatments. These results suggest that the visual and vocal components of tidbitting are redundant, but they also highlight the importance of a temporal dimension for any categorization scheme. Visual displays also evoked inspection behavior, characterized by close binocular fixation on the head of the playback male, which is known to facilitate individual recognition. This may also allow hens to assess male quality. Such social responses reveal that tidbitting probably has multiple functions and provide a new insight into the selective factors responsible for the evolution of this complex multimodal signal.


Disentangling the influence of environmental and anthropogenic factors on the distribution of endemic vascular plants in Sardinia.
Due to the impelling urgency of plant conservation and the increasing availability of high resolution spatially interpolated (e.g. climate variables) and categorical data (e.g. land cover and vegetation type), many recent studies have examined relationships among plant species distributions and a diversified set of explanatory factors; nevertheless, global and regional patterns of endemic plant richness remain in many cases unexplained. One such pattern is the 294 endemic vascular plant taxa recorded on a 1 km resolution grid on the environmentally heterogeneous island of Sardinia. Sixteen predictors, including topographic, geological, climatic and anthropogenic factors, were used to model local (number of taxa inside each 1 km grid cell) Endemic Vascular Plant Richness (EVPR). Generalized Linear Models were used to evaluate how each factor affected the distribution of local EVPR. Significant relationships with local EVPR and topographic, geological, climatic and anthropogenic factors were found. In particular, elevation explained the larger fraction of variation in endemic richness but other environmental factors (e.g. precipitation seasonality and slope) and human-related factors (e.g. the Human Influence Index (HII) and the proportion of anthropogenic land uses) were, respectively, positively and negatively correlated with local EVPR. Regional EVPR (number of endemic taxa inside each 100 m elevation interval) was also measured to compare local and regional EVPR patterns along the elevation gradient. In contrast to local, regional EVPR tended to decrease with altitude partly due to the decreasing area covered along altitude. The contrasting results between local and regional patterns suggest that local richness increases as a result of increased interspecific aggregation along altitude, whereas regional richness may depend on the interaction between area and altitude. This suggests that the shape and magnitude of the species-area relationship might vary with elevation. This work provides-for the first time in Sardinia-a comprehensive analysis of the influence of environmental factors on the pattern of EVPR in the entire territory, from sea level to the highest peaks. Elevation, as well as other environmental and human-related variables, were confirmed to be influencing factors. In addition, variations of EVPR patterns at regional-to-local spatial scales inspire next investigations on the possible interaction between elevation and area in explaining patterns of plant species richness.


Creativity and oedipal fantasy in Austen's Emma: 'An ingenious and animating suspicion'.
Austen's Emma is one of the great novels of the Western tradition. In this paper the author explores the meaning of Emma's 'ingenious and animating suspicion' that Jane Fairfax seduced her best friend's husband, Mr Dixon. The interpretation that a psychoanalytic understanding makes possible shows how this suspicion represents an oedipal fantasy projected on to Miss Fairfax. Further exploration demonstrates how the fantasy is linked both to Emma's systematic unkindness to Jane Fairfax and to Emma's famous insult to Jane's aunt, Miss Bates. Emma's suspicion projects an oedipal fantasy with its incestuous impulses on to her rival and satisfies an envious aggression at the same time. The author's purpose in this paper is to bring to light through psychoanalytic understanding Austen's dramatisation of the complexity and creativity of the oedipal situation. In addition to the regression in oedipal fantasy, the primary process also functions with a progressive quality that expands and enriches the ego, a double movement described in Keats's 'negative capability', which has been elaborated by Bion. The primal-scene fantasies are often brought alive in the analytic transference. These situations and painful emotions are dramatically portrayed through Austen's genius as vehicles for change. A sudden integration follows a phase of disorganization: 'It darted through her with the speed of an arrow. Mr Knightley must marry no-one but herself'. Emma, who is Austen's 'imaginist', moves from the projected fantasy of the sad love triangle through envy aggression and the narcissistic blows of self-doubt and loss of love to moments of illumination and connection.


The effect of compressive loading magnitude on in situ chondrocyte calcium signaling.
Chondrocyte metabolism is stimulated by deformation and is associated with structural changes in the cartilage extracellular matrix (ECM), suggesting that these cells are involved in maintaining tissue health and integrity. Calcium signaling is an initial step in chondrocyte mechanotransduction that has been linked to many cellular processes. Previous studies using isolated chondrocytes proposed loading magnitude as an important factor regulating this response. However, calcium signaling in the intact cartilage differs compared to isolated cells. The purpose of this study was to investigate the effect of loading magnitude on chondrocyte calcium signaling in intact cartilage. We hypothesized that the percentage of cells exhibiting at least one calcium signal increases with increasing load. Fully intact rabbit femoral condyle and patellar bone/cartilage samples were incubated in calcium-sensitive dyes and imaged continuously under compressive loads of 10-40 % strain. Calcium signaling was primarily associated with the dynamic loading phase and greatly increased beyond a threshold deformation of about 10 % nominal tissue strain. There was a trend toward more cells exhibiting calcium signaling as loading magnitude increased (p = 0.133). These results provide novel information toward identifying mechanisms underlying calcium-dependent signaling pathways related to cartilage homeostasis and possibly the onset and progression of osteoarthritis.