Optimal planning target volume for stage I testicular seminoma: A Medical Research Council randomized trial. Medical Research Council Testicular Tumor Working Group.
To compare relapse rates and toxicity associated with para-aortic (PA) strip or PA and ipsilateral iliac lymph node irradiation (dogleg [DL] field) (30 Gy/15 fractions/3 weeks) for stage I testicular seminoma. Between July 1989 and May 1993, 478 men with testicular seminoma stage I (T1 to T3; no ipsilateral inguinoscrotal operation before orchiectomy) were randomized (PA, 236 patients; DL, 242 patients). Median follow-up time is 4.5 years. Eighteen relapses, nine in each treatment group, have occurred 4 to 35 months after radiotherapy; among these, four were pelvic relapses, all occurring after PA radiotherapy. However, the 95% confidence interval (CI) for the difference in pelvic relapse rates excludes differences of more than 4%. The 3-year relapse-free survival was 96% (95% CI, 94% to 99%) after PA radiotherapy and 96.6% (95% CI, 94% to 99%) after DL (difference, 0.6%; 95% confidence limits, -3.4%, +4.6%). One patient (PA field) has died from seminoma. Survival at 3 years was 99.3% for PA and 100% for DL radiotherapy. Acute toxicity (nausea, vomiting, leukopenia) was less frequent and less pronounced in patients in the PA arm. Within the first 18 months of follow-up, the sperm counts were significantly higher after PA than after DL irradiation. In patients with testicular seminoma stage I (T1 to T3) and with undisturbed lymphatic drainage, adjuvant radiotherapy confined to the PA lymph nodes is associated with reduced hematologic, gastrointestinal, and gonadal toxicity, but with a higher risk of pelvic recurrence, compared with DL radiotherapy. The recurrence rate is low with either treatment. PA radiotherapy is recommended as standard treatment in these patients.


Chemically modified poly(2-hydroxyethyl methacrylate) cryogel for the adsorption of heparin.
Various clinical procedures, such as cardiovascular surgery or extracorporeal blood purification, involve systemic anticoagulation using heparin. High concentrations of circulating heparin require neutralization due to possible serious bleeding complications. The intravenous administration of the heparin antagonist protamine sulfate is routinely clinically performed, but is frequently associated with adverse reactions. Therefore, there is a need for a valid and safe alternative to achieve extracorporeal heparin removal from blood or plasma, such as a filter, a matrix, or an adsorbent. Here, we describe the development of a macroporous poly(2-hydroxyethyl methacrylate)-based monolithic cryogel functionalized with l-lysine (pHEMA-lys) and the characterization of its selective heparin adsorption. The maximum binding capacity was quantified in vitro using aqueous and serum solutions under static and dynamic conditions, and fresh human plasma under static conditions. The pHEMA-lys bound 40,500 IU and 32,500 IU heparin/g cryogel at the equilibrium in aqueous solution and 50% serum, respectively. In human plasma spiked with 100 IU/mL of heparin, the binding was still highly efficient (4330 IU/g cryogel after 30 min, i.e., 87% of the initial concentration). The cryogels showed good blood compatibility, as indicated by negligible adsorption of albumin, antithrombin III, and total protein, and may thus be suitable for extracorporeal heparin removal.


Management and outcome of acute coronary syndrome patients in relation to prior history of atrial fibrillation.
The prognostic impact of atrial fibrillation (AF) in the setting of acute coronary syndrome (ACS) is controversial. Furthermore, there are limited real-world data on the management of ACS patients with history of AF. The Global Registry of Acute Coronary Events (GRACE/GRACE2) and Canadian Registry of Acute Coronary Events (CANRACE) enrolled 14,285 patients across Canada between 1999 and 2008. Patients were stratified by the presence of history of AF. We compared clinical characteristics, medical therapies, cardiac procedures, and clinical outcomes between the 2 groups. Overall, 1333 of the enrolled patients (9.3%) had history of AF, of whom 51.5% presented with non-ST-segment elevation myocardial infarction, 29.5% with unstable angina, and 19.1% with ST-segment elevation myocardial infarction. Compared with the group without, patients with a history of AF less frequently received evidence-based antiplatelet and antithrombin therapies, left ventricle ejection fraction assessment, and coronary angiography (all P < 0.001); they also had higher unadjusted rates of in-hospital death, myocardial (re)infarction, and heart failure. However, in multivariable analysis, history of AF was not found to be independently associated with in-hospital mortality (adjusted odds ratio [OR] = 1.12; 95% confidence interval (CI), 0.73-1.73; P = 0.61) or death and/or myocardial reinfarction (adjusted OR = 1.15; 95% CI, 0.87-1.5; P = 0.34). History of AF is common among ACS patients. They received less evidence-based medical and invasive therapies than ACS patients without history of AF. History of AF is a negative independent predictor of in-hospital coronary angiography but was not found to be independently associated with adverse outcomes.


Function-focused care programme for older people in Korean long-term care facilities.
This study aimed to develop and test the effect of the Korean Function-Focused Care Programme (K-FFCP) on cognition and physical and psychological status in long-term care facilities' (LTCs) residents. A quasi-experimental repeated-measures design was used. The K-FFCP was developed to maintain and maximise functions of older people in LTCs. After implementing the K-FFCP for six weeks, differences in the cognitive status, activities of daily living (ADL) performance, physical capability, grip strength, fear of falling, depression and anxiety status of the two groups were examined using repeated-measures analysis of variance. Patient outcomes were evaluated at baseline and at 6 and 12 weeks after the intervention. In the experimental group, a significant group-time interaction effect was reported for ADL performance and depression. The current findings provided evidence of its feasibility and indicated significant improvement in older people's functions using the K-FFCP. Although this study is a pilot test, these results may confirm the importance of the K-FFCP for the maintenance of ADL performance in older people in LTCs.


Update on viral hepatitis: 2005.
This review aims to provide a concise summary of relevant developments in the treatment and prevention of viral hepatitis based on publications between December 2004 and November 2005. Long-term therapy with lamivudine was shown to reduce the incidence of adverse clinical outcomes in patients with chronic hepatitis B. This benefit was, however, diminished in patients with viral breakthrough. Combination therapy (peginterferon or telbivudine plus lamivudine) decreased the risk of antiviral resistance but did not improve response compared with monotherapy. Entecavir and tenofovir were shown to be effective in suppressing lamivudine-resistant hepatitis B. A shorter duration of peginterferon-ribavirin treatment was shown to be feasible in genotype 2/3 hepatitis C patients. Relapsers to previous standard interferon-based therapy had higher sustained virological response rates than previous non-responders when retreated with peginterferon-ribavirin. Hepatitis C patients with advanced liver disease may achieve sustained virological response with peginterferon-ribavirin treatment, but the complication rate was high. New insights into the management of viral hepatitis have been gained over the past year. The problem of antiviral resistance in chronic hepatitis B is real and treatment options need to keep evolving. Advances have been made in hepatitis C treatment including patients with genotypes 2 or 3 infection and decompensated cirrhosis.


Protective effects of in vivo-expressed autotransporters against Bordetella pertussis infection.
Bordetella pertussis causes whooping cough, a severe and prolonged respiratory disease that results inhas high morbidity and mortality rates, particularly in developing countries. The number incidence of whooping cough cases is increasing in many countries despite high vaccine coverage. Causes for the re-emergence of the disease include the limited duration of protection conferred by the acellular pertussis vaccines (aP)s and pathogenic adaptations that involve antigenic divergence from vaccine strains. Therefore, current vaccines therefore need to be improved. In the present study, we focused on five autotransporters: namely SphB1, BatB, SphB2, Phg, and Vag8, which were previously found to be expressed by B. bronchiseptica during the course of infection in rats and examined their protective efficiencies as vaccine antigens. The passenger domains of these proteins were produced in recombinant forms and used as antigens. An intranasal murine challenge assay showed that immunization with a mixture of SphB1 and Vag8 (SV) significantly reduced bacterial load in the lower respiratory tract and a combination of aP and SV acts synergistically in effects of conferring protection against B. pertussis infection, implying that these antigens have potential as components to for improvinge th the currently available acellular pertussis vaccine.


Fusogenic membrane glycoprotein-mediated tumour cell fusion activates human dendritic cells for enhanced IL-12 production and T-cell priming.
Fusogenic membrane glycoproteins (FMG) are a family of viral genes that, when expressed in tumour cells, trigger extensive cell to cell fusion and subsequent cell death. Gene therapy approaches using FMG are also potentially immunogenic, since syncitia generated ex vivo can be therapeutic as antitumour vaccines in murine models. This study has addressed the mechanisms responsible for the immunogenicity of FMG-mediated cell death, and its applicability to human immune priming. We show that fusion of human Mel888 melanoma cells following transfection with FMG can reverse the suppressive effects of Mel888 on dendritic cells (DC) phenotype, and potentiate IL-12 production by DC on activation in a cell contact-dependent manner. DC loaded with fusing, but not intact, tumour cells primed a naive, tumour-specific cytotoxic T-cell response, which was MHC class I-restricted and associated with production of high levels of IFNgamma and, later, IL-5. Fusing cells were an effective source of antigen for DC cross-priming and presentation of the melanoma-specific antigen gp100 to a specific T-cell clone. These data show, in a human system, that FMG represent an immunogenic, as well as cytotoxic, gene therapy for cancer, reversing the inhibitory effects of tumour cells on DC to potentiate IL-12 production and naive T-cell priming.


On the cardiac contractile, electrophysiological and biochemical effects of endothall, a protein phosphatase inhibitor.
Protein phosphatase inhibitors, e.g. cantharidin, exert positive inotropic effects in mammalian heart preparations. Endothall, a synthetic herbicide which is chemically related to cantharidin, inhibits protein phosphatase activities in mouse liver preparations. However, the cardiac effects of endothall have hitherto not been studied. In guinea pig papillary muscles, endothall (1-100 micromol/l) failed to affect force of contraction, whereas cantharidin (1-100 micromol/l) increased force of contraction maximally to 313.4 +/- 32% of control at 10 micromol/l. In isolated guinea pig ventricular cardiomyocytes, endothall did neither change the free intracellular calcium concentration nor the amplitude of calcium current nor the phosphorylation state of regulatory phosphoproteins like phospholamban. In contrast, cantharidin (30 micromol/l) increased the free intracellular calcium concentration and the L-type calcium current to 149.6 +/- 9% and to 157.6 +/- 12% of control, respectively. Furthermore, cantharidin (1-100 micromol/l) augmented the phosphorylation of phospholamban maximally to 140.8 +/- 7% of control. Nevertheless, in guinea pig ventricular homogenates, both endothall and cantharidin inhibited phosphatase activity with EC(50) values of 1.92 and 0.32 micromol/l, respectively. Thus, in contrast to cantharidin, endothall failed to increase force of contraction, though it inhibited protein phosphatase activity. Clearly, endothall is not an appropriate tool to study the function of protein phosphatases in the mammalian heart.


Mushroom consumption, biomarkers, and risk of cardiovascular disease and type 2 diabetes: a prospective cohort study of US women and men.
Mushrooms are good dietary sources of important vitamins, minerals, and bioactive compounds which may be important in the prevention of chronic diseases. However, studies have not prospectively evaluated the potential health effects of mushrooms with respect to major cardiometabolic diseases. The aim of this study was to examine the association of mushroom consumption with major cardiometabolic diseases and mediating biomarkers in 2 large prospective US cohorts. We followed 67,139 women from the Nurses' Health Study (1986-2012) and 43,541 men from the Health Professionals Follow-up Study (1986-2012) who were free of chronic diseases. Mushroom consumption was assessed at baseline through the use of a food-frequency questionnaire. Cardiometabolic biomarkers were collected in subpopulations of the 2 cohorts. Cox proportional hazards models were used to estimate HRs and 95% CIs of cardiovascular disease (CVD), including coronary heart disease (CHD) and stroke, and type 2 diabetes (T2D), associated with mushroom consumption. We identified total 11,894 CVD (7,616 CHD; 4,278 stroke), and 10,206 T2D cases in >2 million person-years of follow-up. In the pooled multivariable-adjusted analysis, participants who consumed ≥5 servings of mushrooms per week had no significantly different risk of total CVD (HR: 1.02; 95% CI: 0.91, 1.14), CHD (HR: 1.00; 95% CI: 0.87, 1.16), stroke (HR: 1.05; 95% CI: 0.87, 1.25), or T2D (HR: 1.04; 95% CI: 0.93, 1.16) than participants who consumed mushrooms <1 time/mo. We consistently found no association between mushroom consumption and the aforementioned cardiometabolic diseases, in subgroups of sex, lifestyle factors, and medical conditions. Moreover, mushroom consumption was not associated with plasma biomarkers of lipids, insulin, and inflammation. We found no association of mushroom consumption with biomarkers and risks of CVD and T2D in US adults. More large prospective cohort studies are warranted to investigate this association in other racial/ethnic groups.


More Impaired Dynamic Ventilatory Muscle Oxygenation in Congestive Heart Failure than in Chronic Obstructive Pulmonary Disease.
Patients with chronic obstructive pulmonary disease (COPD) and congestive heart failure (CHF) often have dyspnea. Despite differences in primary organ derangement and similarities in secondary skeletal muscle changes, both patient groups have prominent functional impairment. With similar daily exercise performance in patients with CHF and COPD, we hypothesized that patients with CHF would have worse ventilatory muscle oxygenation than patients with COPD. This study aimed to compare differences in tissue oxygenation and blood capacity between ventilatory muscles and leg muscles and between the two patient groups. Demographic data, lung function, and maximal cardiopulmonary exercise tests were performed in 134 subjects without acute illnesses. Muscle oxygenation and blood capacity were measured using frequency-domain near-infrared spectroscopy (fd-NIRS). We enrolled normal subjects and patients with COPD and CHF. The two patient groups were matched by oxygen-cost diagram scores, New York Heart Association functional classification scores, and modified Medical Research Council scores. COPD was defined as forced expired volume in one second and forced expired vital capacity ratio ≤0.7. CHF was defined as stable heart failure with an ejection fraction ≤49%. The healthy subjects were defined as those with no obvious history of chronic disease. Age, body mass index, cigarette consumption, lung function, and exercise capacity were different across the three groups. Muscle oxygenation and blood capacity were adjusted accordingly. Leg muscles had higher deoxygenation (HHb) and oxygenation (HbO2) and lower oxygen saturation (SmO2) than ventilatory muscles in all participants. The SmO2 of leg muscles was lower than that of ventilatory muscles because SmO2 was calculated as HbO2/(HHb+HbO2), and the HHb of leg muscles was relatively higher than the HbO2 of leg muscles. The healthy subjects had higher SmO2, the patients with COPD had higher HHb, and the patients with CHF had lower HbO2 in both muscle groups throughout the tests. The patients with CHF had lower SmO2 of ventilatory muscles than the patients with COPD at peak exercise (p < 0.01). We conclud that fd-NIRS can be used to discriminate tissue oxygenation of different musculatures and disease entities. More studies on interventions on ventilatory muscle oxygenation in patients with CHF and COPD are warranted.


Fast-searching algorithm for vector quantization using projection and triangular inequality.
In this paper, a new and fast-searching algorithm for vector quantization is presented. Two inequalities, one used for terminating the searching process and the other used to delete impossible codewords, are presented to reduce the distortion computations. Our algorithm makes use of a vector's features (mean value, edge strength, and texture strength) to reject many unlikely codewords that cannot be rejected by other available approaches. Experimental results show that our algorithm is superior to other algorithms in terms of computing time and the number of distortion calculations. Compared with available approaches, our method can reduce the computing time and the number of distortion computations significantly. Compared with the best method of reducing distortion computation, our algorithm can further reduce the number of distortion calculations by 29% to 58.4%. Compared with the best encoding algorithm for vector quantization, our approach also further reduces the computing time by 8% to 47.7%.


Intelligent Head-Mounted Obstacle Avoidance Wearable for the Blind and Visually Impaired.
Individuals who are Blind and Visually Impaired (BVI) take significant risks and dangers on obstacles, particularly when they are unaccompanied. We propose an intelligent head-mount device to assist BVI people with this challenge. The objective of this study is to develop a computationally efficient mechanism that can effectively detect obstacles in real time and provide warnings. The learned model aims to be both reliable and compact so that it can be integrated into a wearable device with a small size. Additionally, it should be capable of handling natural head turns, which can generally impact the accuracy of readings from the device's sensors. Over thirty models with different hyper-parameters were explored and their key metrics were compared to identify the most suitable model that strikes a balance between accuracy and real-time performance. Our study demonstrates the feasibility of a highly efficient wearable device that can assist BVI individuals in avoiding obstacles with a high level of accuracy.


Antimotility agents for the treatment of Clostridium difficile diarrhea and colitis.
Antimotility agent use for the treatment of Clostridium difficile infection (CDI) is discouraged. We reviewed the literature and unpublished postmarketing surveillance reports regarding antimotility treatment of CDI. Twenty reports met inclusion criteria, describing 55 patients with CDI who were exposed to antimotility agents. All studies were case reports or series, with the exception of 1 retrospective review. Nineteen patients (35%) improved, with clinical resolution. Nine patients (16%) died, and 27 patients (49%) had unknown outcomes. Seventeen patients (31%) with CDI developed colonic dilation; 5 of these patients with severe CDI died. However, all patients who experienced complications or died were given antimotility agents alone initially, without an appropriate antibiotic. Twenty-three patients who received metronidazole or vancomycin coadministered with the antimotility agent experienced no complications. Evidence supporting the hypothesis that worsened outcomes are associated with antimotility therapy of CDI is lacking. Further study of the role of antimotility agents in providing symptomatic relief and reducing environmental contamination with infectious stool may be warranted.


Variability of mitochondrial subgenomic molecules in the meristematic cells of higher plants.
MtDNAs from BY-2 cells and rice root were analyzed by random amplified polymorphic DNA (RAPD) assay and Southern hybridization analysis. A number of differences were observed in the RAPD patterns amplified from mtDNAs sampled at different phases of the BY-2 cell culture. RAPD fragments also varied with the template DNAs derived from various areas of rice root tip. When a RAPD fragment was hybridized to restriction fragments of whole DNAs, isolated from the distal area of the apical meristem and differentiated elongation zone of a root, two distinct stoichiometric differences were observed in the hybridization signals. This suggests that the organization of mt-genome in prototypic cells in the root apical meristem differs from that found in the differentiated cells.


[State of the coronary circulation in patients with a chronic aneurysm of the heart].
Examination covered 56 patients with chronic post-infarction aneurysm of the heart. It included electro- and phonocariography, selective coronarography and ventriculography of the left heart. In the majority of these patients the affection started with myocardial infarction which gave rise to the development of the cardiac aneurysm. Coronarography disclosed a lesion of the left anterior descending artery in the form of an extended occlusion. Collateral coronary circulation was indefinable. Ventriculography of the left heart disclosed an aneurysm whose localization accorded with electrocardiographic indications. In the development of aneurysms of the heart the authors attach great importance to presence of collateral circulation at the instant of an acute lesion of the coronary artery.


Comparison of Early and Late Latent Syphilis- Colorado, 1991.
Latent syphilis (i.e., the presence of serological evidence for syphilis without clinical manifestations) is divided into early latent ([EL]<l-year's duration) and late latent ([LL] more than 1-year's duration) stages.1 LL syphilis, which is often associated with low nontreponemal test (e.g., rapid plasma reagin [RPR]) titers and is presumed to have been acquired in the distant past, is not routinely included in syphilis surveillance reports and analyses. Although a separate classification of "unknown latent syphilis" has been proposed,1 in practice, duration is unknown for nearly all syphilis cases that are classified as LL. This report compares EL and LL syphilis cases in Colorado during 1991 and demonstrates substantial overlap in their characteristics.Colorado EL and LL syphilis cases reported in 1991 were abstracted for information on age, sex, racial/ethnic group, and serologic test results (RPR). Persons aged≥60 years with RPR titers≤16 were not included among LL cases, because these.


[Effect of Panax notoginseng saponins on intercellular adhesion molecule-1 expression and neutrophil infiltration in cerebral infarction tissue of rats].
To investigate the effect of Panax notoginseng saponins (Pns) on the expression of intercellular adhesion molecule-1 (ICAM-1) and brain neutrophil infiltration induced by transient focal cerberal ischemia and reperfusion in rats. Focal cerebral ischemia-reperfusion model in rats (male) were made by transient occlusion of the middle cerebral artery (MCA) for 2 h followed by 24 h reperfusion. Pns 25 and 50 mg/kg were administered ip at 2 h after reperfusion. At 24 h after reperfusion, neutrophil infiltration and the expression of intercellular adhesion molecule-1 (ICAM-1) were measured with myeloperoxidase (MPO) activity, and immunohistochemistry, respectively. After ischemia reperfusion, the expression of ICAM-1 and MPO activity in the ischemic hemisphere cortex of MCA area and caudate putamen were markedly increased. Treatment with 50 mg/kg Pns inhibited neutrophil infiltration and the expression of ICAM-1. Pns inhibited neutrophil infiltration and the expression of ICAM-1 following focal cerebral ischemia-reperfusion in rats, attenuated the ischemic brain inflammation.


Modelling Dictyostelium discoideum morphogenesis: the culmination.
The culmination of the morphogenesis of the cellular slime mould Dictyostelium discoideum involves complex cell movements which transform a mound of cells into a globule of spores on a slender stalk. We show that cyclic AMP signalling and differential adhesion, combined with cell differentiation and slime production, are sufficient to produce the morphogenetic cell movements which lead to culmination. We have simulated the process of culmination using a hybrid cellular automata/partial differential equation model. With our model we have been able to reproduce the main features that occur during culmination, namely the straight downward elongation of the stalk, its anchoring to the substratum and the formation of the long thin stalk topped by the spore head. We conclude that the cyclic AMP signalling system is responsible for the elongation and anchoring of the stalk, but in a roundabout way: pressure waves that are induced by the chemotaxis towards cyclic AMP squeeze the stalk through the cell mass. This mechanism forces the stalk to elongate precisely in the direction opposite to that of the chemotactically moving cells. The process turns out to be 'guided' by inactive 'pathfinder' cells, which form the tip of the stalk. We show that the entire development is enacted by means of the aforementioned building blocks. This means that no global gradients or different modes of chemotaxis are needed to complete the culmination.


Evidence for a decrease in cardiovascular risk factors following recombinant growth hormone administration in abstinent anabolic-androgenic steroid users.
To determine whether six days recombinant human growth hormone (rhGH) in an abstinent anabolic-androgenic steroid (AAS) group had any cardiovascular and biochemical effects compared with a control group. Male subjects (n=48) were randomly divided, using a single blind procedure into two groups: (1) control group (C) n=24, mean+/-SD, age 32+/-11 years; height 1.8+/-0.06m; (2) rhGH using group (0.058IUkg(-1)day(-1)) (GH) n=24, mean+/-SD, age 32+/-9 years; height 1.8+/-0.07m. Physiological responses, anthropometry, arterial pulse wave velocity (APWV), blood pressure (BP), heart rate (HR), peak oxygen uptake (VO(2) peak) and biochemical indices were investigated. Body mass index, fat-free mass index and VO(2) peak significantly increased while body fat significantly decreased within GH (all P<0.017). Insulin like growth factor-I significantly increased within GH (P<0.017) and compared with C (P<0.05). Serum sodium significantly increased (P<0.017) and serum homocysteine, high sensitivity C-reactive protein, thyroid stimulating hormone and tetra-iodothyronine (T(4)), significantly decreased within GH (all P<0.017). T(4) significantly decreased compared with C (P<0.05). Arterial pulse wave velocity, peak and recovery systolic and diastolic BP, significantly decreased compared with C (P<0.05). Resting HR and rate pressure product (RPP) significantly increased compared with C (P<0.05). The findings of this study suggest that short term use of rhGH may have beneficial effects on endothelial function and specific inflammatory markers of cardiovascular disease in abstinent AAS users, but may have an adverse effect on the cardiovascular system, as evidenced by the increase in resting RPP.


Transient expression of Mnb/Dyrk1a couples cell cycle exit and differentiation of neuronal precursors by inducing p27KIP1 expression and suppressing NOTCH signaling.
The decision of a neural precursor to stop dividing and begin its terminal differentiation at the correct place, and at the right time, is a crucial step in the generation of cell diversity in the nervous system. Here, we show that the Down's syndrome candidate gene (Mnb/Dyrk1a) is transiently expressed in prospective neurons of vertebrate CNS neuroepithelia. The gain of function (GoF) of Mnb/Dyrk1a induced proliferation arrest. Conversely, its loss of function (LoF) caused over proliferation and cell death. We found that MNB/DYRK1A is both necessary and sufficient to upregulate, at transcriptional level, the expression of the cyclin-dependent kinase inhibitor p27(KIP1) in the embryonic chick spinal cord and mouse telencephalon, supporting a regulatory role for MNB/DYRK1A in cell cycle exit of vertebrate CNS neurons. All these actions required the kinase activity of MNB/DYRK1A. We also observed that MNB/DYRK1A is co-expressed with the NOTCH ligand Delta1 in single neuronal precursors. Furthermore, we found that MNB/DYRK1A suppressed NOTCH signaling, counteracted the pro-proliferative action of the NOTCH intracellular domain (NICD), stimulated Delta1 expression and was required for the neuronal differentiation induced by the decrease in NOTCH signaling. Nevertheless, although Mnb/Dyrk1a GoF led to extensive withdrawal of neuronal precursors from the cell cycle, it was insufficient to elicit their differentiation. Remarkably, a transient (ON/OFF) Mnb/Dyrk1a GoF efficiently induced neuronal differentiation. We propose that the transient expression of MNB/DYRK1A in neuronal precursors acts as a binary switch, coupling the end of proliferation and the initiation of neuronal differentiation by upregulating p27KIP1 expression and suppressing NOTCH signaling.


Influence of Co-Dosed Lipids from Biota Extracts on the Availability of Chemicals in In Vitro Cell-Based Bioassays.
Extraction of chemicals from biota leads to co-extraction of lipids. When dosing such extracts into in vitro bioassays, co-dosed lipids act as an additional phase that can reduce the bioavailability of the chemicals and the apparent sensitivity of the assay. Equilibrium partitioning between medium, cells, and co-dosed lipids was described with an existing equilibrium partitioning model for cell-based bioassays extended by an additional lipid phase. We experimentally investigated the influence of co-dosed lipids on the effects elicited by four test chemicals of different hydrophobicity in two bioassays, indicative of the aryl hydrocarbon receptor and oxidative stress response (AREc32). The partitioning model explained the effect of the test chemicals in the presence of spiked triolein within a factor of 0.33-5.83 between the measured and predicted effect concentration (EC). We applied the model to marine mammal blubber extracted with silicone. Extracts dosed in the AREc32 bioassay showed a linear increase of apparent EC with increasing lipid fraction. The partitioning model was used to interpret the role of the co-extracted lipid. A quantitative lipid correction of bioassay results in the presence of co-dosed lipids was possible for known compounds and defined mixtures, while we could only estimate a range for mixtures of unknown chemicals.


The Convolutional Group Sequential Test: Reducing Test Time for Evoked Potentials.
When using a statistical test for automatically detecting evoked potentials, the number of stimuli presented to the subject (the sample size for the statistical test) should be specified at the outset. For evoked response detection, this may be inefficient, i.e., because the signal-to-noise ratio (SNR) of the response is not known in advance, the user would usually err on the cautious side and use a relatively high number of stimuli to ensure adequate statistical power. A more efficient approach is to apply the statistical test repeatedly to the accumulating data over time, as this allows the test to be stopped early for the high SNR responses (thus reducing test time), or later for the low SNR responses. The caveat is that the critical decision boundaries for rejecting the null hypothesis need to be adjusted if the intended type-I error rate is to be obtained. This study presents an intuitive and flexible method for controlling the type-I error rate for sequentially applied statistical tests. The method is built around the discrete convolution of truncated probability density functions, which allows the null distribution for the test statistic to be constructed at each stage of the sequential analysis. Because the null distribution remains tractable, the procedure for finding the stage-wise critical decision boundaries is greatly simplified. The method also permits data-driven adaptations (using data from previous stages) to both the sample size and the statistical test, which offers new opportunities to speed up testing for evoked response detection.


The first case of Plasmodium ovale malaria from Bangladesh.
The authors report the first indigenous case of Plasmodium ovale infection from Bangladesh. The diagnosis was confirmed by PCR and sequence analysis. The patient had neither been outside of the country nor ever received blood transfusions. The authors concluded that there was evidence for a local transmission of P ovale malaria in Bangladesh. P ovale malaria should therefore always be considered a potential differential diagnosis in the indigenous population as well as travellers and migrants returning from South Asia, possibly up to years after their return.


Implementing Precision Medicine and Artificial Intelligence in Plastic Surgery: Concepts and Future Prospects.
Precision medicine, or the individualization of evidence-based medicine, is forthcoming. As surgeons, we must be prepared for the integration of patient and system factors. Plastic surgeons regard themselves as innovators and early adopters. As such, we need our adaptability now more than ever to implement digital advancements and precision medicine into our practices. The integration of artificial intelligence (AI) technology and the capture of big data techniques should foster the next great leaps in medicine and surgery, allowing us to capture the detailed minutiae of precision medicine. The algorithmic process of artificial neural networks will guide large-scale analysis of data, including features such as pattern recognition and rapid quantification, to organize and distribute data to surgeons seamlessly. This vast digital collection of information, commonly termed "big data," is only one potential application of AI. By incorporating big data, the cognitive abilities of a surgeon can be complemented by the computer to improve patient-centered care. Furthermore, the use of AI will provide individual patients with increased access to the broadening world of precision medicine. Therefore, plastic surgeons must learn how to use AI within the contexts of our practices to keep up with an evolving field in medicine. Although rudimentary in its practice, we present a glimpse of the potential applications of AI in plastic surgery to incorporate the practice of precision medicine into the care that we deliver.


Localization of gene and protein expressions of tumor necrosis factor-{alpha} and tumor necrosis factor receptor types I and II in the bovine corpus luteum during the estrous cycle.
One of the many roles of tumor necrosis factor (TNF)-α is to control mammalian corpus luteum (CL) PG synthesis and apoptotic cell death. Here, the cellular localization of TNF-α and its type I (TNF-RI) and type II (TNF-RII) receptors in bovine luteal tissue were analyzed using in situ hybridization, immunohistochemistry, and quantitative real-time PCR. Transcripts for TNF-α were expressed in bovine CL throughout the estrous cycle, but were significantly more abundant (P < 0.01) at the regressed luteal stage than at the other stages. Localization of TNF-α transcripts and protein were observed in large and small bovine luteal cells, as well as in immune cells. Moreover, transcripts for TNF-RI and TNF-RII were expressed in bovine CL throughout the estrous cycle. The abundance of TNF-RII transcripts was greater (P < 0.01) at the regressed luteal stage than at the other stages, whereas TNF-RI transcript abundance did not significantly change. Expression of TNF-RI and TNF-RII transcripts and proteins were observed in both the large and small luteal cells, and the proteins were also expressed in the immune cells and vascular endothelial cells. These results suggest that TNF-α sources include immune cells, as well as large and small luteal cells, and that TNF-RI and TNF-RII are present in the luteal cells of the bovine CL.


Recent evidence for an expanded role of the kynurenine pathway of tryptophan metabolism in neurological diseases.
The kynurenine pathway (KP) of tryptophan metabolism has emerged in recent years as a key regulator of the production of both neuroprotective (e.g. kynurenic and picolinic acid, and the essential cofactor NAD+) and neurotoxic metabolites (e.g. quinolinic acid, 3-hydroxykynurenine). The balance between the production of the two types of metabolites is controlled by key rate-limiting enzymes such as indoleamine-2,3-dioxygenase (IDO-1), and in turn, molecular signals such as interferon-γ (IFN-γ), which activate the KP metabolism of tryptophan by this enzyme, as opposed to alternative pathways for serotonin and melatonin production. Dysregulated KP metabolism has been strongly associated with neurological diseases in recent years, and is the subject of increasing efforts to understand how the metabolites are causative of disease pathology. Concurrent with these endeavours are drug development initiatives to use inhibitors to block certain enzymes in the pathway, resulting in reduced levels of neurotoxic metabolites (e.g. quinolinic acid, an excitotoxin and N-Methyl-d-Aspartate (NMDA) receptor agonist), while in turn enhancing the bioavailability of the neuroprotective metabolites such as kynurenic acid. Neurodegenerative diseases often have a substantial autoimmune or inflammatory component; hence a greater understanding of how KP metabolites influence the inflammatory cascade is required. Additionally, challenges exist in diseases like multiple sclerosis (MS) and motor neurone disease (MND), which do not have reliable biomarkers. Clinical diagnosis can often be prolonged in order to exclude other diseases, and often diagnosis occurs at an advanced state of disease pathology, which does not allow a lengthy time for patient assessment and intervention therapies. This review considers the current evidence for involvement of the KP in several neurological diseases, in biomarkers of disease and also the parallels that exist in KP metabolism with what is known in other diseases such as HIV, Alzheimer's disease/dementia, infection, immune privilege and cardiovascular disease. This article is part of the Special Issue entitled 'The Kynurenine Pathway in Health and Disease'.


Gender-dependent bladder response to one-day bladder outlet obstruction.
Development of bladder fibrosis, loss of compliance, and voiding dysfunction are among the severe consequences of various lower urinary conditions, for example, bladder outlet obstruction (BOO), neurogenic bladder, and radiotherapy to the pelvic area. The bladder remodelling results in significant changes in bladder function and architecture, and may ultimately be deleterious for kidney function. The molecular signals underlying pathologic bladder remodelling, as well as the impact of gender, remain poorly understood. To investigate the bladder remodelling after one day BOO, whether the remodelling is different between different bladder sections, and whether genders may affect the remodelling. Thirty male and 30 female C57BL/6NRj mice were randomly divided into Control, Sham and BOO groups with ten mice per group. A 24-h total urethral obstruction was performed at the proximal urethra. Histological changes were observed via H&E, trichrome and immunohistochemistry staining. Harvested bladders were divided into upper and lower sections for analysis. Protein and gene expression were detected by Western blotting and qPCR. No significant changes in bladder wall thickness were observed following BOO, while increased bladder mass after BOO was found in female mice only. We detected FN and ⍺-SMA upregulation in the male upper bladder segment. Female BOO mice bladders showed increased α-SMA expression in both bladder segments, but no difference of FN was observed in either bladder segments. BOO-induced upregulation of TGF-β and Gremlin were detected in both male and female bladders, while downregulation of BMP-7 was detected only in male bladders. Furthermore, phosphorylation of both SMAD2/3 and SMAD1/5/9 were increased in male bladders following BOO, whereas female mice exhibited increased pSMAD2/3 in the upper and increased pSMAD1/5/9 in the lower bladder segment. Our data indicate that some specific proteins and growth factors were detected as early alterations of tissue which may lead to fibrosis. In addition, the males tended to have more pronounced response than females. However, the causes and consequences of the findings need to be further investigated.


Real-time modal control implementation for adaptive optics.
The electronics, computing hardware, and computing used to provide real-time modal control for a laser guide-star adaptive optics system are presented. This approach offers advantages in the control of unobserved modes, the elimination of unwanted modes (e.g., tip and tilt), and automatic handling of the case of low-resolution lens arrays. In our two-step modal implementation, the input vector of gradients is first decomposed into a Zernike polynomial mode by a least-squares estimate. The number of modes is assumed to be less than or equal to the number of actuators. The mode coefficients are then available for collection and analysis or for the application of modal weights. Thus the modal weights may be changed quickly without recalculating the full matrix. The control-loop integrators are at this point in the algorithm. To calculate the deformable-mirror drive signals, the mode coefficients are converted to the zonal signals by a matrix multiply. When the number of gradients measured is less than the number of actuators, the integration in the control loop will be done on the lower-resolution grid to avoid growth of unobserved modes. These low-resolution data will then be effectively interpolated to yield the deformable-mirror drive signals.


Rheuma-VOR study: optimising healthcare of rheumatic diseases by multiprofessional coordinating centres.
Early diagnosis of inflammatory arthritis is critical to prevent joint damage and functional incapacities. However, the discrepancy between recommendations of early diagnosis and reality is remarkable. The Rheuma-VOR study aimed to improve the time to diagnosis of patients with early arthritis by coordinating cooperation between primary care physicians, specialists and patients in Germany. This prospective non-randomised multicentre study involved 2340 primary care physicians, 72 rheumatologists, 4 university hospitals and 4 rheumatology centres in 4 German Federal States. The two coprimary endpoints (time to diagnosis and screening performance of primary care physicians) were evaluated for early versus late implementation phase. Additionally, time to diagnosis and secondary endpoints (decrease of disease activity, increase in quality of life and overall well-being, improvement of fatigue, depression, functional ability, and work ability, reduction in drug and medical costs and hospitalisation) were compared with a reference cohort of the German Rheumatism Research Centre (DRFZ) reflecting standard care. A total of 7049 patients were enrolled in the coordination centres and 1537 patients were diagnosed with a rheumatic disease and consented to further participation. A follow-up consultation after 1 year was realised in 592 patients. The time to diagnosis endpoint and the secondary endpoints were met. In addition, the calculation of cost-effectiveness shows that Rheuma-VOR has a dominant cost-benefit ratio compared with standard care. Rheuma-VOR has shown an improvement in rheumatological care, patient-reported outcome parameters and cost savings by coordinating the cooperation of primary care physicians, rheumatologists and patients, in a nationwide approach.


Comparison of Dexmedetomidine-Ketamine-Midazolam and Isoflurane for Anesthesia of Black-tailed Prairie Dogs (Cynomys ludovicianus).
Few studies evaluate anesthesia in black-tailed prairie dogs (Cynomys ludovicianus). Isoflurane inhalant anesthesia is used in this species most commonly, but injectable protocols are poorly described. Here we compared the physiologic effects, including anesthetic depth, vital signs, and hematologic changes, of anesthetic protocols using isoflurane or a combination of dexmedetomidine, ketamine, and midazolam in black-tailed prairie dogs. In a randomized, complete crossover study design, intact male black-tailed prairie dogs (n = 9; age, 6 mo) were anesthetized by using a combination of dexmedetomidine (0.25 mg/kg IM), ketamine (40 mg/kg IM), and midazolam (1.5 mg/kg IM). For reversal, atipamezole (0.15 mg/kg) and flumazenil (0.05 mg/kg) were administered 45 min after induction. For comparison, isoflurane was administered at 5% in 100% oxygen at 5 L/min in an anesthetic induction chamber, followed by maintenance isoflurane 2% in 2 L/min oxygen through a tight-fitting facemask for 45 min. Induction and recovery time, respiratory rate, heart rate, body temperature, SpO₂, indirect blood pressure, and reflexes were monitored every 5 min during the anesthetic period. Blood samples for venous blood gases, PCV, and refractometric total protein were obtained from the cranial vena cava at 5 min and 45 min. Both protocols appeared to achieve safe and effective anesthesia. Except for blood pressure, all vital signs differed between the 2 treatments. Isoflurane anesthesia resulted in a slightly longer induction and lower respiratory rate and body temperature but increased likelihood of absent reflexes. DKM anesthesia resulted in a faster induction and less hypothermia but also prolonged recovery and lower heart rate and SpO₂ readings. These findings suggest that isoflurane provides a more stable and consistent anesthetic plane, whereas dexmedetomidine-ketamine-midazolam anesthesia may be an effective alternative for short procedures that require fast induction and limited analgesia.


A BAYESIAN SPATIAL AND TEMPORAL MODELING APPROACH TO MAPPING GEOGRAPHIC VARIATION IN MORTALITY RATES FOR SUBNATIONAL AREAS WITH R-INLA.
Hierarchical Bayes models have been used in disease mapping to examine small scale geographic variation. State level geographic variation for less common causes of mortality outcomes have been reported however county level variation is rarely examined. Due to concerns about statistical reliability and confidentiality, county-level mortality rates based on fewer than 20 deaths are suppressed based on Division of Vital Statistics, National Center for Health Statistics (NCHS) statistical reliability criteria, precluding an examination of spatio-temporal variation in less common causes of mortality outcomes such as suicide rates (SRs) at the county level using direct estimates. Existing Bayesian spatio-temporal modeling strategies can be applied via Integrated Nested Laplace Approximation (INLA) in R to a large number of rare causes of mortality outcomes to enable examination of spatio-temporal variations on smaller geographic scales such as counties. This method allows examination of spatiotemporal variation across the entire U.S., even where the data are sparse. We used mortality data from 2005-2015 to explore spatiotemporal variation in SRs, as one particular application of the Bayesian spatio-temporal modeling strategy in R-INLA to predict year and county-specific SRs. Specifically, hierarchical Bayesian spatio-temporal models were implemented with spatially structured and unstructured random effects, correlated time effects, time varying confounders and space-time interaction terms in the software R-INLA, borrowing strength across both counties and years to produce smoothed county level SRs. Model-based estimates of SRs were mapped to explore geographic variation.


Iron entry in neurons and astrocytes: a link with synaptic activity.
Iron plays a fundamental role in the development of the central nervous system (CNS) as well as in several neuronal functions including synaptic plasticity. Accordingly, neuronal iron supply is tightly controlled: it depends not only on transferrin-bound iron but also on non-transferrin-bound iron (NTBI), which represents a relevant quote of the iron physiologically present in the cerebrospinal fluid (CSF). Different calcium permeable channels as well as the divalent metal transporter 1 (DMT1) have been proposed to sustain NTBI entry in neurons and astrocytes even though it remains an open issue. In both cases, it emerges that the control of iron entry is tightly linked to synaptic activity. The iron-induced oxidative tone can, in physiological conditions, positively influence the calcium levels and thus the synaptic plasticity. On the other hand, an excess of iron, with the ensuing uncontrolled production of reactive oxygen species (ROS), is detrimental for neuronal survival. A protective mechanism can be played by astrocytes that, more resistant to oxidative stress, can uptake iron, thereby buffering its concentration in the synaptic environment. This competence is potentiated when astrocytes undergo activation during neuroinflammation and neurodegenerative processes. In this minireview we focus on the mechanisms responsible for NTBI entry in neurons and astrocytes and on how they can be modulated during synaptic activity. Finally, we speculate on the relevance they may have in both physiological and pathological conditions.


Synergistic effects of 2A-mediated polyproteins on the production of lignocellulose degradation enzymes in tobacco plants.
Cost-effective bioethanol production requires a supply of various low-cost enzymes that can hydrolyse lignocellulosic materials consisting of multiple polymers. Because plant-based enzyme expression systems offer low-cost and large-scale production, this study simultaneously expressed β-glucosidase (BglB), xylanase (XylII), exoglucanase (E3), and endoglucanase (Cel5A) in tobacco plants, which were individually fused with chloroplast-targeting transit peptides and linked via the 2A self-cleaving oligopeptideex from foot-and-mouth disease virus (FMDV) as follows: [RsBglB-2A-RaCel5A], [RsXylII-2A-RaCel5A], and [RsE3-2A-RaCel5A]. The enzymes were targeted to chloroplasts in tobacco cells and their activities were confirmed. Similarly to the results of a transient assay using Arabidopsis thaliana protoplasts, when XylII was placed upstream of the 2A sequence, the [RsXylII-2A-RaCel5A] transgenic tobacco plant had a more positive influence on expression of the protein placed downstream. The [RsBglB-2A-RaCel5A] and [RsE3-2A-RaCel5A] transgenic lines displayed higher activities towards carboxylmethylcellulose (CMC) compared to those in the [RsXylII-2A-RaCel5A] transgenic line. This higher activity was attributable to the synergistic effects of the different cellulases used. The [RsBglB-2A-RaCel5A] lines exhibited greater efficiency (35-74% increase) of CMC hydrolysis when the exoglucanase CBHII was added. Among the various exoglucanases, E3 showed higher activity with the crude extract of the [RsBglB-2A-RaCel5A] transgenic line. Transgenic expression of 2A-mediated multiple enzymes induced synergistic effects and led to more efficient hydrolysis of lignocellulosic materials for bioethanol production.


Clustering Based on Periodicity in High-Throughput Time Course Data.
Nuclear magnetic resonance (NMR) spectroscopy, traditionally used in analytical chemistry, has recently been introduced to studies of metabolite composition of biological fluids and tissues. Metabolite levels change over time, and providing a tool for better extraction of NMR peaks exhibiting periodic behavior is of interest. We propose a method in which NMR peaks are clustered based on periodic behavior. Periodic regression is used to obtain estimates of the parameter corresponding to period for individual NMR peaks. A mixture model is then used to develop clusters of peaks, taking into account the variability of the regression parameter estimates. Methods are applied to NMR data collected from human blood plasma over a 24-hour period. Simulation studies show that the extra variance component due to the estimation of the parameter estimate should be accounted for in the clustering procedure.


Annotation of gene promoters by integrative data-mining of ChIP-seq Pol-II enrichment data.
Use of alternative gene promoters that drive widespread cell-type, tissue-type or developmental gene regulation in mammalian genomes is a common phenomenon. Chromatin immunoprecipitation methods coupled with DNA microarray (ChIP-chip) or massive parallel sequencing (ChIP-seq) are enabling genome-wide identification of active promoters in different cellular conditions using antibodies against Pol-II. However, these methods produce enrichment not only near the gene promoters but also inside the genes and other genomic regions due to the non-specificity of the antibodies used in ChIP. Further, the use of these methods is limited by their high cost and strong dependence on cellular type and context. We trained and tested different state-of-art ensemble and meta classification methods for identification of Pol-II enriched promoter and Pol-II enriched non-promoter sequences, each of length 500 bp. The classification models were trained and tested on a bench-mark dataset, using a set of 39 different feature variables that are based on chromatin modification signatures and various DNA sequence features. The best performing model was applied on seven published ChIP-seq Pol-II datasets to provide genome wide annotation of mouse gene promoters. We present a novel algorithm based on supervised learning methods to discriminate promoter associated Pol-II enrichment from enrichment elsewhere in the genome in ChIP-chip/seq profiles. We accumulated a dataset of 11,773 promoter and 46,167 non-promoter sequences, each of length 500 bp, generated from RNA Pol-II ChIP-seq data of five tissues (Brain, Kidney, Liver, Lung and Spleen). We evaluated the classification models in building the best predictor and found that Bagging and Random Forest based approaches give the best accuracy. We implemented the algorithm on seven different published ChIP-seq datasets to provide a comprehensive set of promoter annotations for both protein-coding and non-coding genes in the mouse genome. The resulting annotations contain 13,413 (4,747) protein-coding (non-coding) genes with single promoters and 9,929 (1,858) protein-coding (non-coding) genes with two or more alternative promoters, and a significant number of unassigned novel promoters. Our new algorithm can successfully predict the promoters from the genome wide profile of Pol-II bound regions. In addition, our algorithm performs significantly better than existing promoter prediction methods and can be applied for genome-wide predictions of Pol-II promoters.


Differentiation and Characterization of Neural Progenitors and Neurons from Mouse Embryonic Stem Cells.
We describe the step-by-step procedure for culturing and differentiating mouse embryonic stem cells into neuronal lineages, followed by a series of assays to characterize the differentiated cells. The E14 mouse embryonic stem cells were used to form embryoid bodies through the hanging drop method, and then induced to differentiate into neural progenitor cells by retinoic acid, and finally differentiated into neurons. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence experiments revealed that the neural progenitors and neurons exhibit corresponding markers (nestin for neural progenitors and neurofilament for neurons) at day 8 and 12 post-differentiation, respectively. Flow cytometry experiments on an E14 line expressing a Sox1 promoter-driven GFP reporter showed that about 60% of cells at day 8 are GFP positive, indicating the successful differentiation of neural progenitor cells at this stage. Finally, RNA-seq analysis was used to profile the global transcriptomic changes. These methods are useful for analyzing the involvement of specific genes and pathways in regulating the cell identity transition during neuronal differentiation.


Studies on the interactions of SAP-1 (an N-terminal truncated form of cystatin S) with its binding partners by CD-spectroscopic and molecular docking methods.
SAP-1 is a 113 amino acid long single-chain protein which belongs to the type 2 cystatin gene family. In our previous study, we have purified SAP-1 from human seminal plasma and observed its cross-class inhibitory property. At this time, we report the interaction of SAP-1 with diverse proteases and its binding partners by CD-spectroscopic and molecular docking methods. The circular dichroism (CD) spectroscopic studies demonstrate that the conformation of SAP-1 is changed after its complexation with proteases, and the alterations in protein secondary structure are quantitatively calculated with increase of α-helices and reduction of β-strand content. To get insight into the interactions between SAP-1 and proteases, we make an effort to model the three-dimensional structure of SAP-1 by molecular modeling and verify its stability and viability through molecular dynamics simulations and finally complexed with different proteases using ClusPro 2.0 Server. A high degree of shape complementarity is examined within the complexes, stabilized by a number of hydrogen bonds (HBs) and hydrophobic interactions. Using HB analyses in different protein complexes, we have identified a series of key residues that may be involved in the interactions between SAP-1 and proteases. These findings will assist to understand the mechanism of inhibition of SAP-1 for different proteases and provide intimation for further research.


That "poker face" just might lose you the game! The impact of expressive suppression and mimicry on sensitivity to facial expressions of emotion.
Successful interpersonal functioning often requires both the ability to mask inner feelings and the ability to accurately recognize others' expressions--but what if effortful control of emotional expressions impacts the ability to accurately read others? In this study, we examined the influence of self-controlled expressive suppression and mimicry on facial affect sensitivity--the speed with which one can accurately identify gradually intensifying facial expressions of emotion. Muscle activity of the brow (corrugator, related to anger), upper lip (levator, related to disgust), and cheek (zygomaticus, related to happiness) were recorded using facial electromyography while participants randomized to one of three conditions (Suppress, Mimic, and No-Instruction) viewed a series of six distinct emotional expressions (happiness, sadness, fear, anger, surprise, and disgust) as they morphed from neutral to full expression. As hypothesized, individuals instructed to suppress their own facial expressions showed impairment in facial affect sensitivity. Conversely, mimicry of emotion expressions appeared to facilitate facial affect sensitivity. Results suggest that it is difficult for a person to be able to simultaneously mask inner feelings and accurately "read" the facial expressions of others, at least when these expressions are at low intensity. The combined behavioral and physiological data suggest that the strategies an individual selects to control his or her own expression of emotion have important implications for interpersonal functioning.


Depurination of Brome mosaic virus RNA3 in vivo results in translation-dependent accelerated degradation of the viral RNA.
Pokeweed antiviral protein (PAP) is a ribosome-inactivating protein isolated from the pokeweed plant (Phytolacca americana) that exhibits antiviral activity against several plant and animal viruses. We have shown previously that PAP depurinates Brome mosaic virus (BMV) RNAs in vitro and that prior incubation of these RNAs with PAP reduced their synthesis in barley protoplasts. To investigate the post-transcriptional effect of PAP on viral RNA in vivo, we transcribed BMV RNA3 and expressed PAP in the yeast, Saccharomyces cerevisiae, which is a surrogate host for BMV. With an inducible transcription system, we show that the half-life of RNA3 in PAP-expressing cells was significantly less than in cells expressing PAPx, its enzymatically inactive form. PAP bound to RNA3 and depurinated the RNA within open reading frames 3 and 4 and within untranslated regions of the RNA. The depurinated RNA was associated with polysomes, caused ribosomes to stall at the point of depurination, and was targeted for accelerated degradation by components of the No-go decay pathway. As a consequence of translation elongation arrest and increased RNA degradation, expression of PAP in yeast also decreased the level of protein 3a, encoded by the 5'-proximal open reading frame 3 of BMV RNA3. These data provide the first evidence of viral RNA depurination in vivo by any ribosome-inactivating protein and support our hypothesis that depurination contributes to the antiviral activity of PAP, by enhancing viral RNA degradation and reducing translation of viral protein product.


Knowledge, attitudes and behaviours of caregivers regarding children's exposure to environmental tobacco smoke among Arabic and Vietnamese-speaking communities in Sydney, Australia.
The aim of this study was to inform the design of a culturally appropriate health communication campaign addressing exposure of young children to Environmental Tobacco Smoke (ETS) in homes and cars, targeted at caregivers of children aged 0-6 years in the Arabic and Vietnamese-speaking communities in Sydney, Australia. Nine focus groups (five Arabic and four Vietnamese) were conducted by qualified bilingual facilitators, recorded, transcribed, then translated and analysed qualitatively. Results revealed concerning levels of knowledge, attitudes and behaviours regarding children's exposure to ETS. A poor understanding existed regarding what constituted passive smoking and the severity and extent of harm that ETS has on children. Smoking attitudes and behaviours described were not always consistent with the concern expressed about children's vulnerability to smoke. Initial changes in smoking patterns at home since the arrival of children were not always sustained. Despite awareness of the harm caused by smoking in the presence of their children, some caregivers continued to smoke in closed environments. Others felt it was sufficient protection to smoke in another room, with doors and/or windows open or by ensuring that visible smoke did not reach the child directly, whilst some smoked in the home or car only when children were not present. Smoking attitudes and behaviours in the presence of children were influenced by cultural and social norms associated with smoking. The desire to be sociable and hospitable often superseded concerns for children's health. The findings suggest that an effective communication campaign focusing on ETS and children in the Arabic and Vietnamese-speaking communities in Sydney should attempt to address knowledge deficits regarding ETS, as well as tackling entrenched culturally related attitudes and behaviours associated with smoking which are strongly linked to sociability and hospitality.


The role of gender as a barrier to the professional development of psychiatrists.
Despite efforts toward greater gender equality in clinical and academic psychiatry in recent years, more information is needed about the challenges in professional development within psychiatry, and how these may vary with gender. A cross-sectional 27-item online survey was conducted with psychiatrists and psychiatric trainee members of the European Psychiatric Association. A total of 561 psychiatrists and psychiatric trainees from 35 European countries participated representing a response rate of 52.8% for women and 17.7% for men from a total sample of 1,580. The specific challenges that women face in their professional development fall into two categories. One comprised women's negative attitudes concerning their abilities in self-promotion and networking. The other identified environmental barriers related to lack of opportunity and support and gender discrimination. Compared to men, women reported higher rates of gender discrimination in terms of professional advancement. Women were less likely to agree that their institutions had regular activities promoting inclusion, diversity, and training to address implicit gender bias. Working in high-income countries compared to middle-income countries relates to reporting institutional support for career progression. These findings are an open call to hospital leaders, deans of medical schools, and department chairs to increase efforts to eradicate bias against women and create safer, inclusive, and respectful environments for all psychiatrists, a special call to women psychiatrists to be aware of inner tendencies to avoid self-promotion and networking and to think positively and confidently about themselves and their abilities.


Tumour fatty acid metabolism in the context of therapy resistance and obesity.
Fatty acid metabolism is known to support tumorigenesis and disease progression as well as treatment resistance through enhanced lipid synthesis, storage and catabolism. More recently, the role of membrane fatty acid composition, for example, ratios of saturated, monounsaturated and polyunsaturated fatty acids, in promoting cell survival while limiting lipotoxicity and ferroptosis has been increasingly appreciated. Alongside these insights, it has become clear that tumour cells exhibit plasticity with respect to fatty acid metabolism, responding to extratumoural and systemic metabolic signals, such as obesity and cancer therapeutics, to promote the development of aggressive, treatment-resistant disease. Here, we describe cellular fatty acid metabolic changes that are connected to therapy resistance and contextualize obesity-associated changes in host fatty acid metabolism that likely influence the local tumour microenvironment to further modify cancer cell behaviour while simultaneously creating potential new vulnerabilities.


Genomic variation of an endosymbiotic dinoflagellate (Symbiodinium 'fitti') among closely related coral hosts.
Mutualisms where hosts are coupled metabolically to their symbionts often exhibit high partner fidelity. Most reef-building coral species form obligate symbioses with a specific species of photosymbionts, dinoflagellates in the family Symbiodiniaceae, despite needing to acquire symbionts early in their development from environmental sources. Three Caribbean acroporids (Acropora palmata, A. cervicornis and their F1 hybrid) are sympatric across much of their range, but often occupy different depth and light habitats. Throughout this range, both species and their hybrid associate with the endosymbiotic dinoflagellate Symbiodinium 'fitti'. Because light (and therefore depth) influences the physiology of dinoflagellates, we investigated whether S. 'fitti' populations from each host taxon were differentiated genetically. Single nucleotide polymorphisms (SNPs) among S. 'fitti' strains were identified by aligning shallow metagenomic sequences of acroporid colonies sampled from across the Caribbean to a ~600-Mb draft assembly of the S. 'fitti' genome (from the CFL14120 A. cervicornis metagenome). Phylogenomic and multivariate analyses revealed that genomic variation among S. 'fitti' strains partitioned to each host taxon rather than by biogeographical origin. This is particularly noteworthy because the hybrid has a sparse fossil record and may be of relatively recent origin. A subset (37.6%) of the SNPs putatively under selection were nonsynonymous mutations predicted to alter protein efficiency. Differences in genomic variation of S. 'fitti' strains from each host taxon may reflect the unique selection pressures created by the microenvironments associated with each host. The nonrandom sorting among S. 'fitti' strains to different hosts could be the basis for lineage diversification via disruptive selection, leading to ecological specialization and ultimately speciation.


Does drinking pattern modify the effect of alcohol on the risk of coronary heart disease? Evidence from a meta-analysis.
To evaluate the strength of the evidence provided by epidemiological literature investigating drinking pattern as effect modifier of alcohol intake on the risk of coronary heart disease (CHD). Meta-analysis of observational studies. Medline, citation tracking, from 1966 to 2006. Original studies investigating the amount of alcohol intake, combined with the frequency of alcohol consumption and/or pattern of alcohol drinking affecting the risk of CHD were extracted. Among them, cohort and case-control studies reporting sufficient data to perform statistical analyses and using people who abstained from alcohol as the reference were included. Six (4 cohort and 2 case-control) out of 118 studies reviewed met the inclusion criteria. Compared with those who abstained from alcohol, regular heavy drinkers and heavy irregular or binge drinkers showed significantly different pooled relative risks of 0.75 (95% confidence interval 0.64 to 0.89) and 1.10 (1.03 to 1.17) respectively. The dose-response relation between the amount of alcohol intake and CHD risk was significantly different in regular and irregular drinkers. A J-shaped curve, with nadir around 28 grams of alcohol per week, and last protective dose of 131 grams per week, was obtained including drinkers who consumed alcohol for 2 days a week or less. Conversely, in people who consumed alcohol for more than 2 days a week a significant protective effect was seen even when drinking high amounts of alcohol. This meta-analysis suggests that binge and heavy irregular drinking modify the favourable effect of alcohol intake on the CHD risk. However, this conclusion should be taken with caution because of the small number of studies considered.


Acetazolamide and human carbonic anhydrases: retrospect, review and discussion of an intimate relationship.
Acetazolamide (AZM) is a strong pharmacological sulphonamide-type (R-SO2-NH2, pKa 7.2) inhibitor of the activity of several carbonic anhydrase (CA) isoforms, notably of renal CA II (Ki, 12 nM) and CA IV (Ki, 74 nM). AZM is clinically used for about eighty years in various diseases including epilepsy and glaucoma. Pharmacological AZM increases temporarily the urinary excretion of bicarbonate (HCO3-) and sodium ions (Na+) and sustainably the urinary pH. AZM is excreted almost unchanged over several hours at high rates in the urine. Closely parallel concentrations of circulating and excretory AZM are observed upon administration of therapeutical doses of AZM. In a proof-of-principle study, we investigated the effects of the ingestion of a 250-mg AZM-containing tablet by a healthy volunteer on the urinary excretion of organic and inorganic substances over 5 h (range, 0, 0.5, 1, 1.5, 2, 3, 4, 5 h). Measured analytes included: AZM, amino acids and their metabolites such as guanidinoacetate, i.e. the precursor of creatine, of asymmetrically (ADMA) and symmetrically (SDMA) dimethylated arginine, nitrite (O = N-O-, pKa 3.4) and nitrate (O2N-O-, pKa -1.37), the major metabolites of nitric oxide (NO), the C-H acidic malondialdehyde (MDA; (CHO)2CH2, pKa 4.5), and creatinine for correction of analytes excretion. All analytes were measured by validated isotopologues using gas chromatography-mass spectrometry (GC-MS) methods. AZM excretion in the urine reached its maximum value after 2 h and was fairly stable for the next 3 h. Time series analysis by the ARIMA method was performed. AZM ingestion increased temporarily the urinary excretion of the amino acids Leu + Ile, nitrite and nitrate, decreased temporarily the urinary excretion of other amino acids. AZM decreased sustainably the urinary excretion of MDA, a biomarker of oxidative stress (i.e. lipid peroxidation). Whether this decrease is due to inhibition of the excretion of MDA or attenuation of oxidative stress by AZM is unknown. The acute and chronic effects of AZM on the urinary excretion of electrolytes and physiological substances reported in the literature are discussed in depth in the light of its extraordinary pharmacokinetics and pharmacodynamics. Tolerance development/drug resistance to AZM in chronic use and potential mechanisms are also addressed.


Comparing polybrominated diphenyl ethers (PBDEs) in airborne particles in Guangzhou and Hong Kong: sources, seasonal variations and inland outflow.
The historical application/usage and management of chemicals in Hong Kong have been distinctively different from mainland China. In the present study, polybrominated diphenyl ethers (PBDEs) were measured in year-round atmospheric particle samples collected from urban Hong Kong and Guangzhou for comparison. The concentrations of BDE-209 and Sigma9PBDEs (defined as the sum of BDE-28, -47, -66, -100, -99, -154, -153, -138 and -183) in Guangzhou ranged from 758 to 21,900 pg m(-3) and from 31.8 to 3320 pg m(-3), respectively, and in Hong Kong ranged from 8.5 to 895 pg m(-3) and from 1.0 to 386 pg m(-3), respectively. Elevated concentrations of PBDEs were observed in Guangzhou, showing significant atmospheric PBDE pollution. BDE-209, -47, and -99 were the dominant congeners in all the samples, suggesting that the widely used commercial penta- and deca-BDE products were the original sources. Distinct seasonal patterns were observed in the PBDE concentrations of aerosols in Hong Kong, higher during the winter monsoon period, and lower during summertime. The less distinct seasonal variations of PBDE concentrations in the aerosols of Guangzhou suggested the dominance of local pollution sources around the city. Significant correlations were found between BDE-209 and organic carbon (OC) or elemental carbon (EC) in the two cities, suggesting that combustion may be an important pathway introducing BDE-209 to the atmosphere. The lower BDE-209 concentrations along with higher OC/EC ratios implied that a quick loss of BDE-209 may occur during the aerosol aging processes. Back trajectory analysis showed that the high PBDE concentrations observed in Hong Kong may be related to the outflows from the inland area of the Pearl River Delta (PRD) by prevailing the northeast or northwest wind in winter.


Benchmarking the GW Approximation and Bethe-Salpeter Equation for Groups IB and IIB Atoms and Monoxides.
Energies from the GW approximation and the Bethe-Salpeter equation (BSE) are benchmarked against the excitation energies of transition-metal (Cu, Zn, Ag, and Cd) single atoms and monoxide anions. We demonstrate that best estimates of GW quasiparticle energies at the complete basis set limit should be obtained via extrapolation or closure relations, while numerically converged GW-BSE eigenvalues can be obtained on a finite basis set. Calculations using real-space wave functions and pseudopotentials are shown to give best-estimate GW energies that agree (up to the extrapolation error) with calculations using all-electron Gaussian basis sets. We benchmark the effects of a vertex approximation (ΓLDA) and the mean-field starting point in GW and the BSE, performing computations using a real-space, transition-space basis and scalar-relativistic pseudopotentials. While no variant of GW improves on perturbative G0W0 at predicting ionization energies, G0W0ΓLDA-BSE computations give excellent agreement with experimental absorption spectra as long as off-diagonal self-energy terms are included. We also present G0W0 quasiparticle energies for the CuO-, ZnO-, AgO-, and CdO- anions, in comparison to available anion photoelectron spectra.


[A Case of Gastric Cancer Responding to Neoadjuvant Chemotherapy Leading to Histological Change to Grade 3].
A 70-year-old man with cStage III A(cT3N2H0P0CYXM0)advanced gastric cancer in the lesser curvature with esophageal invasion and bulky lymph nodes was treated with S-1/CDDP. After 4 courses of chemotherapy, the tumor and lymph nodes were found to be reduced in a CT examination. Total gastrectomy with lymph node dissection(D2)was performed. Histopathological examination revealed no cancer cells in the stomach or lymph nodes, indicating Grade 3.


Sodium chloride and water transport in the descending limb of Henle.
The unique membrane characteristics of the thin descending limb of Henle (DLH) play an integral part in the operation of the countercurrent system. We examined these properties in vitro by perfusing isolated thin descending limbs of rabbits. Active transport of NaCl was ruled out by failure to demonstrate either net transport or transmembrane potential difference when perfusing with isosmolal ultrafiltrate of the same rabbit serum as the bath. Transmembrane potential was zero, and net fluid transport was -0.07 +/-0.06 nl mm(-1) min(-1), which also is not significantly different from zero. Passive permeability coefficient for Na(P(Na)) was determined from the disappearance rate of (22)Na from isosmolal perfusion solution. P(Na) was surprisingly low, 1.61 +/-0.27 x 10(-5) cm sec(-1), a figure which is significantly less than P(Na) in the proximal convoluted tubule (PCT). Reflection coefficient for NaCl (sigmaNaCl) was measured by perfusing the tubule with Na-free raffinose solution in a bath of rabbit serum to which sufficient NaCl was added to obtain conditions of zero net fluid movement. The measured sigmaNaCl of 0.96 +/-0.01 is significantly greater than sigmaNaCl in the PCT. Water permeability to osmotic gradients (L(p)) was determined by perfusing with ultrafiltrate of rabbit serum in a bath made hyperosmotic by addition of either 100 mOsm raffinose or NaCl. L(p) with raffinose was 1.71 +/-0.15 x 10(-4) ml cm(-2) sec(-1) atm(-1) and with NaCl 1.62 +/-0.05 x 10(-4) ml cm(-2) sec(-1) atm(-1), indicating much greater water permeability than in the PCT. In each case the measured increase in osmolality of the collected fluid was primarily due to water efflux without significant influx of solute. The finding of low permeability to sodium and high permeability to water is consonant with the hypotheses that high interstitial concentration of Na in the medulla generates an effective osmotic pressure which results in concentration of the fluid as it courses through the DLH primarily by abstraction of water without significant net entry of NaCl.


Locality and circulation in the Habsburg empire: disputing the Carlsbad medical salt, 1763-1784.
By looking at the fierce debates in the city of Carlsbad in Bohemia around the fabrication of medical salt by a local doctor, David Becher, from 1763 to 1784, the paper examines the interactions between different spheres or levels of circulation of knowledge in the Habsburg Empire. The dispute crystallized around the definition of the product, about its medical qualities and its relation with the water of the local mineral spring. The city's inhabitants contested the vision of the medical experts, fearing that the extraction of the medical salt from the spring water and its sale outside the town would have a negative effect on the number of visitors to the spa. Their vision implied a more or less 'popularized' form of alchemical thinking as it identified the mineral water with the extracted 'salt', conceived as the 'essence' of the water, produced by evaporation. The Carlsbad salt dispute highlights the complex interactions among the different networks in which knowledge circulated through the Habsburg Empire in the eighteenth century. The different actors relied on specific networks with different logics of discourse and different modes of circulation. In each case the relation between the local, the regional and the imperial had to be negotiated. The paper thus sketches out the different geographies of knowledge in the Habsburg Empire but also its localization in and around Carlsbad.


Plasma osmolality, volume, and renin activity at the "anaerobic threshold".
Plasma renin activity (PRA), volume (PV), osmolality, and hemodynamic parameters were examined in relation to the anaerobic threshold (AT) during progressive cycle ergometry (PE) and repetitive bouts of unilateral isokinetic knee extension-flexion (LE) at 50% maximum voluntary contractions in eight normotensive males. During PE, the observed rise in PRA paralleled that of lactate with abrupt increases occurring at the AT. Correlation of % delta lactate (La), % delta osmolality, and % delta PV with % delta PRA were r = 0.65, 0.36, and -0.51, respectively (all P less than 0.01). In addition, when mean arterial pressure was plotted as a function of VO2, the rate of rise was greater below the AT than above the AT (11.5 vs. -2.4 mmHg X l-1 X min, P less than 0.001). A time control study (TC) exercising subjects for the same duration but at work rates maintained below the AT resulted in significantly lower values for both PRA and La (7.18 vs. 11.27 mg angiotensin I (ANG I) X ml-1 X min and 3.16 vs. 9.93 mM, P less than 0.05 for TC vs. PE) while producing a similar fall in % delta PV and rise in osmolality. During LE, a high correlation was obtained for % delta PRA and % delta La (r = 0.86, P less than 0.01) but not for % delta PRA with % delta PV or % delta osmolality. The data demonstrate that PRA parallels lactate during exercise and that mean arterial pressure rises more slowly beyond the AT despite a more rapid rise in PRA.


[The Oslo96 reform and skills updating practices].
We have previously shown that 2-3 years after completion of their studies, doctors who had followed the study programme Oslo96 spent somewhat more time on updating their skills than other doctors trained at the University of Oslo, but less time than doctors trained at the University of Bergen. The purpose of this study is to investigate whether these differences had changed 5.5-7 years after graduation. We used questionnaire data collected from the last cohort to follow the old programme of study at UiO (graduated 2001), the second cohort to follow the Oslo96 programme (graduated 2003) and the cohort that graduated from the University of Bergen in 2003. Data were collected 5.5-7 years after graduation (T2) and compared to corresponding data collected 2-3 years after graduation (T1). The study included only respondents who had answered all questions at both T1 and T2. The response rate ranged from 55% to 59% in the three samples. At 5.5-7 years after graduation we found no differences with regard to the time spent on updating between Oslo96 doctors (median: 230 minutes/week) and doctors who had graduated before the Oslo96 reform or from the University of Bergen (median: 240 minutes/week in both groups). All three groups reported a statistically significant increase from T1 to T2 in their use of the Internet for purposes of skills updating. Training courses and conferences were the preferred forms of skills updating. A correlation between the programme of study and the doctors' habits with regard to skills updating could not be detected.


An Examination of the Impact of Childhood Emotional Abuse and Gender on Cannabis Use Trajectories among Community Youth.
Cannabis is the most frequently used illicit substance among youth, with rates of cannabis use escalating across adolescence. One potential factor predicting cannabis use among youth is childhood emotional abuse (CEA), which has been associated with substance use behaviors more broadly. Although CEA may be associated with increased cannabis use in general, it is likely that sex may have an impact on these relations, given that girls are more likely to use substances following abuse experiences than boys. The purpose of the current study as to examine longitudinal relations between CEA and gender on cannabis use during adolescence. The current study included a sample of 206 9th grade community youth (120 boys; Mage = 14.10, 55% European-American) followed annually through the 12th grade. CEA was assessed with the Childhood Trauma Questionnaire and cannabis use was assessed with the Youth Risk Behavior Survey. A latent growth model was utilized to examine cannabis use trajectories from grades 9-12. Within our initial model, elevated baseline use was associated with male gender and more severe CEA. Significant predictors of increases in cannabis use over time included elevated baseline alcohol use and the interaction between gender and CEA, such that girls with the most severe CEA had the greatest increases in cannabis use over time. These results suggest the importance of addressing CEA among adolescent girls. Given that cannabis use during adolescence is associated with a host of negative outcomes, targeted efforts to reduce use, through prevention and intervention efforts, is critical.


Vitamin D status and viral response to therapy in hepatitis C infected children.
To study the frequency of vitamin D deficiency in patients with hepatitis C virus (HCV) infection and to evaluate the role of vitamin D supplementation in improving antiviral therapy. Sixty-six children aged from 7-14 years (mean ± SD, 11.17±2.293) diagnosed with HCV infection were matched to 28 healthy controls. Serum levels of 25 (OH) D3, calcium, phosphorus, alkaline phosphatase and plasma level of parathormone were measured. Quantitative PCR for HCV was performed Bone density was determined by dual energy X-ray absorptiometry. All cases received conventional therapy, and only 33 patients received vitamin D supplementation. Children with HCV showed significantly increased levels of HCV RNA (P<0.001), parathormone (P<0.01) and decreased vitamin D levels (P<0.05) (33.3% deficient and 43.3% insufficient) compared with controls. Abnormal bone status (Z score -1.98±0.75) was found in ribs, L-spine, pelvis and total body. Cases treated with vitamin D showed significant higher early (P<0.04) and sustained (P<0.05) virological response. There was a high frequency of vitamin D deficiency among the Egyptian HCV children, with significant decrease in bone density. The vitamin D level should be assessed before the start of antiviral treatment with the correction of any detected deficiency. Adding vitamin D to conventional Peg/RBV therapy significantly improved the virological response and helped to prevent the risk of emerging bone fragility.


The ethics of pharmaceutical research funding: a social organization approach.
This paper advances a social organization approach to examining unethical behavior. While unethical behaviors may stem in part from failures in individual morality or psychological blind spots, they are both generated and performed through social interactions among individuals and groups. To illustrate the value of a social organization approach, a case study of a medical school professor's first experience with pharmaceutical-company-sponsored research is provided in order to examine how funding arrangements can constrain research integrity. The case illustrates three significant ways that institutional corruption can occur in the research process. First, conflicts of norms between pharmaceutical companies, universities, and affiliated teaching hospitals can result in compromises and self-censorship. Second, normal behavior is shaped through routine interactions. Unethical behaviors can be (or can become) normal behaviors when they are produced and reproduced through a network of social interactions. Third, funding arrangements can create networks of dependency that structurally distort the independence of the academic researcher in favor of the funder's interests. More broadly, the case study demonstrates how the social organization approach deepens our understanding of the practice of ethics.


Genetic diversity and population divergences of an indigenous tree (Coffea mauritiana) in Reunion Island: role of climatic and geographical factors.
Oceanic islands are commonly considered as natural laboratories for studies on evolution and speciation. The evolutionary specificities of islands associated with species biology provide unique scenarios to study the role of geography and climate in driving population divergence. However, few studies have addressed this subject in small oceanic islands with heterogeneous climates. Being widely distributed in Reunion Island forest, Coffea mauritiana represents an interesting model case for investigating patterns of within-island differentiation at small spatial scale. In this study, we examined the genetic diversity and population divergences of C. mauritiana using SNP markers obtained from 323 individuals across 34 locations in Reunion Island. Using redundancy analysis, we further evaluated the contribution of geographic and climatic factors to shaping genetic divergence among populations. Genetic diversity analyses revealed that accessions clustered according to the source population, with further grouping in regional clusters. Genetic relationships among the regional clusters underlined a recent process of expansion in the form of step-by-step colonization on both sides of the island. Divergence among source populations was mostly driven by the joint effect of geographic distance and climatic heterogeneity. The pattern of isolation-by-geography was in accordance with the dispersal characteristics of the species, while isolation-by-environment was mostly explained by the heterogeneous rainfall patterns, probably associated with an asynchronous flowering among populations. These findings advance our knowledge on the patterns of genetic diversity and factors of population differentiation of species native to Reunion Island, and will also usefully guide forest management for conservation.


Quantifying and correcting bias in transcriptional parameter inference from single-cell data.
The snapshot distribution of mRNA counts per cell can be measured using single-molecule fluorescence in situ hybridization or single-cell RNA sequencing. These distributions are often fit to the steady-state distribution of the two-state telegraph model to estimate the three transcriptional parameters for a gene of interest: mRNA synthesis rate, the switching on rate (the on state being the active transcriptional state), and the switching off rate. This model assumes no extrinsic noise, i.e., parameters do not vary between cells, and thus estimated parameters are to be understood as approximating the average values in a population. The accuracy of this approximation is currently unclear. Here, we develop a theory that explains the size and sign of estimation bias when inferring parameters from single-cell data using the standard telegraph model. We find specific bias signatures depending on the source of extrinsic noise (which parameter is most variable across cells) and the mode of transcriptional activity. If gene expression is not bursty then the population averages of all three parameters are overestimated if extrinsic noise is in the synthesis rate; underestimation occurs if extrinsic noise is in the switching on rate; both underestimation and overestimation can occur if extrinsic noise is in the switching off rate. We find that some estimated parameters tend to infinity as the size of extrinsic noise approaches a critical threshold. In contrast when gene expression is bursty, we find that in all cases the mean burst size (ratio of the synthesis rate to the switching off rate) is overestimated while the mean burst frequency (the switching on rate) is underestimated. We estimate the size of extrinsic noise from the covariance matrix of sequencing data and use this together with our theory to correct published estimates of transcriptional parameters for mammalian genes.


Microscopic three-dimensional topometry with ferroelectric liquid-crystal-on-silicon displays.
When three dimensional measurements are conducted with fringe projection, the quality of the grating used for the generation of the fringes is important. It has a direct influence on the achievable depth resolution in a given measurement setup. In the past, Ronchi grating or gratings written in nematic liquid-crystal displays or in digital micromirror devices have been used. We report on the application of a reflective ferroelectric liquid-crystal-on-silicon display as the fringe-generating element in a setup based on a stereo microscope. With this device the depth resolution of measurements by use of phase-shifting algorithms can be significantly improved compared with the application of a Ronchi grating or a nematic liquid-crystal display.


Digit regeneration is regulated by Msx1 and BMP4 in fetal mice.
The regeneration of digit tips in mammals, including humans and rodents, represents a model for organ regeneration in higher vertebrates. We had previously characterized digit tip regeneration during fetal and neonatal stages of digit formation in the mouse and found that regenerative capability correlated with the expression domain of the Msx1 gene. Using the stage 11 (E14.5) digit, we now show that digit tip regeneration occurs in organ culture and that Msx1, but not Msx2, mutant mice display a regeneration defect. Associated with this phenotype, we find that Bmp4 expression is downregulated in the Msx1 mutant digit and that mutant digit regeneration can be rescued in a dose-dependent manner by treatment with exogenous BMP4. Studies with the BMP-binding protein noggin show that wild-type digit regeneration is inhibited without inhibiting the expression of Msx1, Msx2 or Bmp4. These data identify a signaling pathway essential for digit regeneration, in which Msx1 functions to regulate BMP4 production. We also provide evidence that endogenous Bmp4 expression is regulated by the combined activity of Msx1 and Msx2 in the forming digit tip; however, we discovered a compensatory Msx2 response that involves an expansion into the wild-type Msx1 domain. Thus, although both Msx1 and Msx2 function to regulate Bmp4 expression in the digit tip, the data are not consistent with a model in which Msx1 and Msx2 serve completely redundant functions in the regeneration response. These studies provide the first functional analysis of mammalian fetal digit regeneration and identify a new function for Msx1 and BMP4 as regulators of the regenerative response.


Critical Role of the Molecular Interface in Double-Layered Pebax-1657/PDMS Nanomembranes for Highly Efficient CO2/N2 Gas Separation.
In this work, we deposited a CO2-selective block copolymer, Pebax-1657, as a selective layer with a thickness of 2-20 nm on the oxygen plasma-activated surface of poly(dimethylsiloxane) (PDMS) used as a gutter layer (thickness ∼400 nm). This double-layered structure was subsequently transferred onto the polyacrylonitrile (PAN) microporous support and studied for CO2/N2 separation. The effect of interfacial molecular arrangements between the selective and gutter layers on CO2 permeance and selectivity has been investigated. We have revealed that the gas permeance and selectivity do not follow the conventional theoretical predictions for the multilayer membrane (resistance in series transport model); specifically, more selective CO2/N2 separation membranes were achieved with ultrathin selective layers. Detailed characterization of the chemical structure of the outermost membrane surface suggests that nanoscale blending of the ultrathin Pebax-1657 layer with O2 plasma-activated PDMS chains on the surface takes place. This nanoblending at the interface between the selective and gutter layers played a critical role in enhancing the CO2/N2 selectivity. CO2 permeances in the developed thin-film composite membranes (TFCM) were between 1200 and 3500 gas permeance units (GPU) and the respective CO2/N2 selectivities were between 72 and 23, providing the gas separation performance suitable for CO2 capture in postcombustion processes. This interpenetrating polymer interface enhanced the overall selectivity of the membrane significantly, exceeding the separation ability of the pristine Pebax-1657 polymer.


Preparation of emulsion hydrogels encapsulating extractant by the Pickering emulsion template method to recover lanthanum ions in aqueous solutions.
To solve the problem of liquid-liquid extraction of La(III), the oil-in-water Pickering emulsions were prepared by utilizing the aqueous solution of sodium alginate as the continuous phase, kerosene-diluted extractant di-(2-ethylhexyl) phosphate (P204) as the dispersed phase, and modified silica as an emulsifier. Then the emulsions were added to a calcium chloride solution to prepare the Pickering emulsion hydrogels (PEHGs) to better remove La(III). The PEHGs were characterized using Fourier transform infrared, thermogravimetric analysis, and scanning electron microscopy. The adsorption properties of PEHGs for La(III) in the aqueous solution were investigated using a UV-vis spectrophotometer. The study found that P204 was successfully coated by hydrogels and reached the highest adsorption capacity of 48 mg/g at pH 4. The amount of adsorption increased with the rise in temperature from 298 to 318 K. La(III) adsorption experimental data were more consistent with the pseudo-second-order kinetic model and the Langmuir isotherm model. Thermodynamic parameters showed that the adsorption of La(III) by PEHGs was a spontaneous endothermic process. The internal diffusion model revealed a linear relationship, indicating that internal diffusion played a role in the adsorption process. The encapsulating property of PEHGs indicated its potential usefulness in industrial wastewater for treating La(III).