IL-6 Receptor Inhibition by Tocilizumab Attenuated Expression of C5a Receptor 1 and 2 in Non-ST-Elevation Myocardial Infarction.
Background: Elevated interleukin-6 (IL-6) and complement activation are associated with detrimental effects of inflammation in coronary artery disease (CAD). The complement anaphylatoxins C5a and C3a interact with their receptors; the highly inflammatory C5aR1, and the C5aR2 and C3aR. We evaluated the effect of the IL-6 receptor (IL-6R)-antagonist tocilizumab on the expression of the anaphylatoxin receptors in whole blood from non-ST-elevation myocardial infarction (NSTEMI) patients. Separately, anaphylatoxin receptor expression in peripheral blood mononuclear cells (PBMC) from patients with different entities of CAD was investigated. Materials and Methods: NSTEMI patients were randomized to one dose of tocilizumab (n = 28) or placebo (n = 32) and observed for 6 months. Whole blood samples drawn at inclusion, at day 2, 3 and after 6 months were used for mRNA isolation. Plasma was prepared for analysis of complement activation measured as sC5b-9 by ELISA. Furthermore, patients with different CAD entities comprising stable angina pectoris (SAP, n = 22), non-ST-elevation acute coronary syndrome (NSTE-ACS, n = 21) and ST-elevation myocardial infarction (STEMI, n = 20) were included. PBMC was isolated from blood samples obtained at admission to hospital and mRNA isolated. Anaphylatoxin-receptor-expression was analyzed with qPCR using mRNA from whole blood and PBMC, respectively. Results: Our main findings were (i) Tocilizumab decreased C5aR1 and C5aR2 mRNA expression significantly (p < 0.001) and substantially (>50%) at day 2 and 3, whereas C3aR expression was unaffected. (ii) Tocilizumab did not affect complement activation. (iii) In analyzes of different CAD entities, C5aR1 expression was significantly increased in all CAD subgroups compared to controls with the highest level in the STEMI patients (p < 0.001). For C5aR2 and C3aR the expression compared to controls were more moderate with increased expression of C5aR2 in the STEMI group (p < 0.05) and C3aR in the NSTE-ACS group (p < 0.05). Conclusion: Expression of C5aR1 and C5aR2 in whole blood was significantly attenuated by IL-6R-inhibition in NSTEMI patients. These receptors were significantly upregulated in PBMC CAD patients with particularly high levels of C5aR1 in STEMI patients.


Short-term effects of large-dose vitamin A supplementation on viral load and immune response in HIV-infected women.
Vitamin A supplementation has been suggested for treatment and prevention of HIV infection. However, some in vitro data indicate that vitamin A may activate HIV. Randomly, 40 HIV-seropositive women of reproductive age were allocated to receive a single oral dose of 9900 micromol (300,000 IU) vitamin A or placebo. Plasma HIV-1 RNA concentration, total lymphocytes, selected lymphocyte subsets and activation markers, and in vitro lymphocyte proliferation to phytohemagglutinin (PHA) and Candida were measured before dosing and at various time points over an 8-week follow-up period. No differences were found between treatment groups in the frequency of signs or symptoms of acute vitamin A toxicity, nor were differences evident in any lymphocyte subset or activation marker at any time during follow-up. Mean and median viral load concentration at each time point and change in viral load from baseline to each follow-up point did not differ between treatment groups. No difference was measured between treatment groups in the proportion of women who responded to PHA or Candida. This study provides no evidence that high dose vitamin A supplementation of HIV-infected women is associated with significant clinical or immunologic adverse effects.


Risk of venous thromboembolism associated with single and combined effects of Factor V Leiden, Prothrombin 20210A and Methylenetethraydrofolate reductase C677T: a meta-analysis involving over 11,000 cases and 21,000 controls.
Genetic and environmental factors interact in determining the risk of venous thromboembolism (VTE). The risk associated with the polymorphic variants G1691A of factor V (Factor V Leiden, FVL), G20210A of prothrombin (PT20210A) and C677T of methylentetrahydrofolate reductase (C677T MTHFR) genes has been investigated in many studies. We performed a pooled analysis of case-control and cohort studies investigating in adults the association between each variant and VTE, published on Pubmed, Embase or Google through January 2010. Authors of eligible papers, were invited to provide all available individual data for the pooling. The Odds Ratio (OR) for first VTE associated with each variant, individually and combined with the others, were calculated with a random effect model, in heterozygotes and homozygotes (dominant model for FVL and PT20210A; recessive for C677T MTHFR). We analysed 31 databases, including 11,239 cases and 21,521 controls. No significant association with VTE was found for homozygous C677T MTHFR (OR: 1.38; 95 % confidence intervals [CI]: 0.98-1.93), whereas the risk was increased in carriers of either heterozygous FVL or PT20210 (OR = 4.22; 95 % CI: 3.35-5.32; and OR = 2.79;95 % CI: 2.25-3.46, respectively), in double heterozygotes (OR = 3.42; 95 %CI 1.64-7.13), and in homozygous FVL or PT20210A (OR = 11.45; 95 %CI: 6.79-19.29; and OR: 6.74 (CI 95 % 2.19-20.72), respectively). The stratified analyses showed a stronger effect of FVL on individuals ≤ 45 years (p value for interaction = 0.036) and of PT20210A in women using oral contraceptives (p-value for interaction = 0.045). In this large pooled analysis, inclusive of large studies like MEGA, no effect was found for C677T MTHFR on VTE; FVL and PT20210A were confirmed to be moderate risk factors. Notably, double carriers of the two genetic variants produced an impact on VTE risk significantly increased but weaker than previously thought.


Proactive coping and gambling disorder among young men.
Objectives Male sex, young age, and frequent gambling are considered as risk factors for gambling disorder (GD) and stress might be one of the triggers of gambling behavior among problem gamblers. Conversely, well-developed coping with stress might counteract gambling problems. The Proactive Coping Theory provides a promising approach for the further development of preventive and treatment measures. The objective of the study was to investigate different facets of proactive coping (PC) in young male gamblers. Methods Young men from Bavaria were recruited via the Munich citizens' registry (n = 2,588) and Facebook invitations (n = 105). In total, 173 out of 398 individuals were positively screened for frequent gambling and/or signs of related problems and completed the baseline questionnaire of the Munich Leisure-time Study. Factors investigated include gambling problems, PC, impulsiveness, social support, and psychological distress. Results Gambling problems were associated with lower levels of preventive coping as well as of adaptive reaction delay. The associations were also significant when controlled for impulsiveness and general psychological distress. Preventive coping moderated the association between social support and gambling problems. Discussion and conclusions Young men with gambling problems less frequently prevent the occurrence of stressors and more often react hasty when these occur. While the investigated group reported good social support, this factor was negatively associated with GD only among individuals with good preventive coping. Preventive coping poses a useful construct for selective prevention and treatment as it can be modified in professional interventions.


Evaluation of macrophage-specific promoters using lentiviral delivery in mice.
In gene therapy, tissue-specific promoters are useful tools to direct transgene expression and improve efficiency and safety. Macrophage-specific promoters (MSPs) have previously been published using different delivery systems. In this study, we evaluated five different MSPs fused with green fluorescent protein (GFP) to delineate the one with highest specificity using lentiviral delivery. We compared three variants of the CD68 promoter (full length, the 343-bp proximal part and the 150-bp proximal part) and two variants (in forward and reverse orientation) of a previously characterized synthetic promoter derived from elements of transcription factor genes. We transduced a number of cell lines and primary cells in vitro. In addition, hematopoietic stem cells were transduced with MSPs and transferred into lethally irradiated recipient mice. Fluorescence activated cell sorting analysis was performed to determine the GFP expression in different cell populations both in vitro and in vivo. We showed that MSPs can efficiently be used for lentiviral gene delivery and that the 150-bp proximal part of the CD68 promoter provides primarily macrophage-specific expression of GFP. We propose that this is the best currently available MSP to use for directing transgene expression to macrophage populations in vivo using lentiviral vectors.


Growth and host interaction of mouse segmented filamentous bacteria in vitro.
The gut microbiota plays a crucial role in the maturation of the intestinal mucosal immune system of its host. Within the thousand bacterial species present in the intestine, the symbiont segmented filamentous bacterium (SFB) is unique in its ability to potently stimulate the post-natal maturation of the B- and T-cell compartments and induce a striking increase in the small-intestinal Th17 responses. Unlike other commensals, SFB intimately attaches to absorptive epithelial cells in the ileum and cells overlying Peyer's patches. This colonization does not result in pathology; rather, it protects the host from pathogens. Yet, little is known about the SFB-host interaction that underlies the important immunostimulatory properties of SFB, because SFB have resisted in vitro culturing for more than 50 years. Here we grow mouse SFB outside their host in an SFB-host cell co-culturing system. Single-celled SFB isolated from monocolonized mice undergo filamentation, segmentation, and differentiation to release viable infectious particles, the intracellular offspring, which can colonize mice to induce signature immune responses. In vitro, intracellular offspring can attach to mouse and human host cells and recruit actin. In addition, SFB can potently stimulate the upregulation of host innate defence genes, inflammatory cytokines, and chemokines. In vitro culturing thereby mimics the in vivo niche, provides new insights into SFB growth requirements and their immunostimulatory potential, and makes possible the investigation of the complex developmental stages of SFB and the detailed dissection of the unique SFB-host interaction at the cellular and molecular levels.


Effects of particle size and annealing on plasmon-induced charge separation at self-assembled gold nanoparticle arrays.
Two-dimensional periodic Au nanoparticle arrays were constructed on TiO2 thin films by a micelle lithography method and seed-mediated photoelectrochemical growth. Their adjustable interparticle distance allows investigation of a particle size effect on plasmon-induced charge separation (PICS) efficiencies without interference from particle aggregation or plasmon coupling. External or internal PICS efficiencies were found to increase and decrease, respectively, with an increase in particle diameter from 25 to 38 nm. Improvement of the contact between Au nanoparticles and TiO2 by annealing enhanced the intensity of a plasmonic interface mode and both external and internal PICS efficiencies.


Two Antarctic penguin genomes reveal insights into their evolutionary history and molecular changes related to the Antarctic environment.
Penguins are flightless aquatic birds widely distributed in the Southern Hemisphere. The distinctive morphological and physiological features of penguins allow them to live an aquatic life, and some of them have successfully adapted to the hostile environments in Antarctica. To study the phylogenetic and population history of penguins and the molecular basis of their adaptations to Antarctica, we sequenced the genomes of the two Antarctic dwelling penguin species, the Adélie penguin [Pygoscelis adeliae] and emperor penguin [Aptenodytes forsteri]. Phylogenetic dating suggests that early penguins arose ~60 million years ago, coinciding with a period of global warming. Analysis of effective population sizes reveals that the two penguin species experienced population expansions from ~1 million years ago to ~100 thousand years ago, but responded differently to the climatic cooling of the last glacial period. Comparative genomic analyses with other available avian genomes identified molecular changes in genes related to epidermal structure, phototransduction, lipid metabolism, and forelimb morphology. Our sequencing and initial analyses of the first two penguin genomes provide insights into the timing of penguin origin, fluctuations in effective population sizes of the two penguin species over the past 10 million years, and the potential associations between these biological patterns and global climate change. The molecular changes compared with other avian genomes reflect both shared and diverse adaptations of the two penguin species to the Antarctic environment.


Colorectal cancer in the north and south of Ireland 1950-1984.
Northern Ireland has the highest standardised mortality ratios for colon cancer in the United Kingdom and the Republic of Ireland has some of the highest mortality rates for cancer in the world. The aim of the study therefore was to investigate trends in colorectal cancer in the north and south of Ireland over the period 1950 to 1984. The study was a cohort analysis of deaths from colorectal cancer for ages 35-74 years by five year age groups, divided by sex. This was a population study involving all cases reported to the Registrar General of Northern Ireland and the Eire Vital Statistics and Central Statistical Office during the study period. As in mainland Britain, rectal cancer mortality declined in the north and the south during the study period, but the fall began sooner for males than females. Colon cancer mortality fell in the late 1950s but subsequently rose to its previous high levels. The observation that there were declines in mortality in the north and south of Ireland in the late 1950s does not support the hypothesis that altered diet due to war rationing in Great Britain and Northern Ireland underlay the fall in British colon cancer mortality after the war. The very high standardised mortality ratios for colon cancer in Northern Ireland highlight a continuing major public health problem in the region.


White-nose syndrome fungus (Geomyces destructans) in bats, Europe.
White-nose syndrome is an emerging disease in North America that has caused substantial declines in hibernating bats. A recently identified fungus (Geomyces destructans) causes skin lesions that are characteristic of this disease. Typical signs of this infection were not observed in bats in North America before white-nose syndrome was detected. However, unconfirmed reports from Europe indicated white fungal growth on hibernating bats without associated deaths. To investigate these differences, hibernating bats were sampled in Germany, Switzerland, and Hungary to determine whether G. destructans is present in Europe. Microscopic observations, fungal culture, and genetic analyses of 43 samples from 23 bats indicated that 21 bats of 5 species in 3 countries were colonized by G. destructans. We hypothesize that G. destructans is present throughout Europe and that bats in Europe may be more immunologically or behaviorally resistant to G. destructans than their congeners in North America because they potentially coevolved with the fungus.


Crystal structure of the termination module of a nonribosomal peptide synthetase.
Nonribosomal peptide synthetases (NRPSs) are modular multidomain enzymes that act as an assembly line to catalyze the biosynthesis of complex natural products. The crystal structure of the 144-kilodalton Bacillus subtilis termination module SrfA-C was solved at 2.6 angstrom resolution. The adenylation and condensation domains of SrfA-C associate closely to form a catalytic platform, with their active sites on the same side of the platform. The peptidyl carrier protein domain is flexibly tethered to this platform and thus can move with its substrate-loaded 4'-phosphopantetheine arm between the active site of the adenylation domain and the donor side of the condensation domain. The SrfA-C crystal structure has implications for the rational redesign of NRPSs as a means of producing novel bioactive peptides.


Laser Doppler flowmetry measurement of changes in human optic nerve head blood flow in response to blood gas perturbations.
The objective of this study was to establish the ability of laser Doppler flowmetry to detect relative changes in human optic nerve head hemodynamics caused by physiologic blood gas perturbations. Laser Doppler flowmetry permits the noninvasive assessment of relative blood velocity, volume, and flow (flux) in a sample volume of the nerve head. Such measurements were performed in two groups of healthy subjects. The first group (n = 11) was tested during normal room air breathing and then while breathing 100% oxygen (isocapnic hyperoxia). The second group (n = 10) was also tested under normal conditions as well as during isoxic hypercapnia (+ 15% end-tidal carbon dioxide). Results were analyzed by paired t tests. Hyperoxia created a significant 25% (p = 0.002) decrease in optic nerve head blood flow, with blood volume decreased by 9% (p = 0.095) and blood velocity reduced by 13% (p = 0.154) compared to the room air condition. During hypercapnia, optic nerve head blood flow was increased by 28% (p = 0.012), with blood volume increased by 22% (p = 0.017) and blood velocity increased by 9% (p = 0.218) as compared to the normal room air condition. Blood flow in the optic nerve head capillaries changes in response to hyperoxia and hypercapnia as demonstrated in the brain and retina. Laser Doppler flowmetry permits the noninvasive assessment of these responses in humans under conditions within the physiologic range.


Visual pigments and the acquisition of visual information.
All the information available to the brain for the interpretation of the visual scene comes from the number of photons absorbed by a very limited number of photoreceptor types which are characterized by their spectral sensitivity. In vertebrates there are considerable differences in the spectral absorption of the rods and cones making up the retinal mosaic of different animals and, in some cases, including fish and primates, there are considerable differences between the cone sets of individuals within a species. Broadly speaking, the spectral sensitivity of the photoreceptors is related to the spectral distribution of the ambient light and this is particularly true of the colour-biased light under water. When an animal migrates from one visual environment to another, its cone complement may change to that suited to the new conditions. However, significant differences between the cone sets of animals living within the same environment and colour vision polymorphism within a species suggest that visual tasks critical to survival or breeding success require particular visual pigment sets. A start has been made in trying to understand what tasks are best served by different pigment sets.


Impact of Complete Revascularization in the ISCHEMIA Trial.
Anatomic complete revascularization (ACR) and functional complete revascularization (FCR) have been associated with reduced death and myocardial infarction (MI) in some prior studies. The impact of complete revascularization (CR) in patients undergoing an invasive (INV) compared with a conservative (CON) management strategy has not been reported. Among patients with chronic coronary disease without prior coronary artery bypass grafting randomized to INV vs CON management in the ISCHEMIA (International Study of Comparative Health Effectiveness with Medical and Invasive Approaches) trial, we examined the following: 1) the outcomes of ACR and FCR compared with incomplete revascularization; and 2) the potential impact of achieving CR in all INV patients compared with CON management. ACR and FCR in the INV group were assessed at an independent core laboratory. Multivariable-adjusted outcomes of CR were examined in INV patients. Inverse probability weighted modeling was then performed to estimate the treatment effect had CR been achieved in all INV patients compared with CON management. ACR and FCR were achieved in 43.4% and 58.4% of 1,824 INV patients. ACR was associated with reduced 4-year rates of cardiovascular death or MI compared with incomplete revascularization. By inverse probability weighted modeling, ACR in all 2,296 INV patients compared with 2,498 CON patients was associated with a lower 4-year rate of cardiovascular death or MI (difference -3.5; 95% CI: -7.2% to 0.0%). In comparison, the event rate difference of cardiovascular death or MI for INV minus CON in the overall ISCHEMIA trial was -2.4%. Results were similar but less pronounced with FCR. The outcomes of an INV strategy may be improved if CR (especially ACR) is achieved. (International Study of Comparative Health Effectiveness With Medical and Invasive Approaches [ISCHEMIA]; NCT01471522).


Aminoalkoxycarbonyloxymethyl Ether Prodrugs with a pH-Triggered Release Mechanism: A Case Study Improving the Solubility, Bioavailability, and Efficacy of Antimalarial 4(1H)-Quinolones with Single Dose Cures.
Preclinical and clinical development of numerous small molecules is prevented by their poor aqueous solubility, limited absorption, and oral bioavailability. Herein, we disclose a general prodrug approach that converts promising lead compounds into aminoalkoxycarbonyloxymethyl (amino AOCOM) ether-substituted analogues that display significantly improved aqueous solubility and enhanced oral bioavailability, restoring key requirements typical for drug candidate profiles. The prodrug is completely independent of biotransformations and animal-independent because it becomes an active compound via a pH-triggered intramolecular cyclization-elimination reaction. As a proof-of-concept, the utility of this novel amino AOCOM ether prodrug approach was demonstrated on an antimalarial compound series representing a variety of antimalarial 4(1H)-quinolones, which entered and failed preclinical development over the last decade. With the amino AOCOM ether prodrug moiety, the 3-aryl-4(1H)-quinolone preclinical candidate was shown to provide single-dose cures in a rodent malaria model at an oral dose of 3 mg/kg, without the use of an advanced formulation technique.


Waveguiding characteristics of surface enhanced Raman scattering signals along crystalline organic semiconducting microrod.
The waveguiding of surface enhanced Raman scattering (SERS) signals was demonstrated by using organic semiconducting microrods (MRs) hybridized with functionalized gold nanoparticles (Au-NPs). Organic semiconducting 1,4-bis(3,5-bis(trifluoromethyl) styryl)-2,5-dibromobenzene (TSDB) crystalline MRs were fabricated as active optical waveguiding system using a self-assembly method. The static SERS effect and the enhancement of photoluminescence were simultaneously observed for the TSDB MRs hybridized with Au-NPs. The waveguiding characteristics of the SERS signals through the hybrid MR of TSDB/Au-NPs were investigated using a high-resolution laser confocal microscope (LCM) system. The enhanced output Raman characteristic modes of TSDB molecules were clearly observed along the hybrid MR of TSDB/Au-NPs, which is attributed to stronger scattering of the light and the increased coupling efficiency of waveguiding due to the presence of Au-NPs. The waveguiding of the SERS signals exhibited different decay constants for the corresponding characteristic Raman modes, such as -C = C- aromatic, -CF3, and C-Br stretching modes. The observed waveguiding characteristics of various SERS modes enable multi-modal waveguiding with relatively narrow spectral resolution for nanophotonic information.


Black Women Are More Likely Than White Women to Schedule a Uterine-Sparing Treatment for Leiomyomas.
Background: To evaluate differences in the proportion of uterine fibroid (UF) treatments that are uterine-sparing between Black women and White women and identify factors that could explain disparities. Methods: Women at age 18-54 years who were enrolled from 10 clinical sites in the United States into the Comparing Options for Management: Patient-Centered Results for UFs (COMPARE-UF) treatment registry completed questionnaires before their UF procedure. UF symptoms and quality of life were assessed by questionnaires. Details on UF imaging and treatment (hysterectomy, myomectomy, or uterine artery embolization [UAE]) were collected from each patient's medical record. Random-effects logistic regression was used to assess the association between race and the odds of having a uterine-sparing procedure versus hysterectomy. Subgroup analyses compared each uterine-sparing procedure with hysterectomy. Results: In this cohort of 1141 White women and 1196 Black women, Black women tended to be younger (median 41.0 vs. 42.0 years) and report worse symptoms, pain, and function on every scale compared with White women. Black women were more likely to have had a prior UF treatment compared with White women (22.8% vs. 14.6%). White women had more hysterectomies (43.6% vs. 32.2%) and myomectomies (50.9% vs. 50.2%) versus Black women. Black women had more UAEs (15.1% vs. 4.7%) than White women. After adjusting for clinical site and other variables, Black women had greater odds than White women of having a myomectomy (odds ratio [OR] = 2.41, 95% confidence interval [CI] = 1.63-3.56) or a UAE versus hysterectomy (OR = 4.24, 95% CI = 2.41-7.46). Conclusion: In these participants, Black women were more likely to schedule a uterine-sparing UF treatment and a nonsurgical UF treatment than their White counterparts; this may not be true for all women. Longer comparative effectiveness studies are needed to inform women about the durability of UF treatments. Greater understanding of factors influencing treatment selection is needed as are studies that include women without access to tertiary care centers. Clinical Trial Registration: Clinicaltrials.gov, NCT02260752 (enrollment start: November 2015).


The effect of regional variations of the trabecular bone properties on the compressive strength of human vertebral bodies.
Cancellous centrum is a major component of the vertebral body and significantly contributes to its structural strength and fracture risk. We hypothesized that the variability of cancellous bone properties in the centrum is associated with vertebral strength. Microcomputed tomography (micro-CT)-based gray level density (GLD), bone volume fraction (BV/TV), and finite element modulus (E) were examined for different regions of the trabecular centrum and correlated with vertebral body strength determined experimentally. Two sets of images in the cancellous centrum were digitally prepared from micro-CT images of eight human vertebral bodies (T10-L5). One set included a cubic volume (1 per vertebral centrum, n = 8) in which the largest amount of cancellous material from the centrum was included but all the shell materials were excluded. The other set included cylindrical volumes (6 per vertebral centrum, n = 48) from the anterior (4 regions: front, center, left, and right of the midline of vertebra) and the posterior (2 regions: left and right) regions of the centrum. Significant positive correlations of vertebral strength with GLD (r (2) = 0.57, p = 0.03) and E (r (2) = 0.63, p = 0.02) of the whole centrum and with GLD (r (2) = 0.65, p = 0.02), BV/TV (r (2) = 0.72, p = 0.01) and E (r (2) = 0.85, p = 0.001) of the central region of the vertebral centrum were found. Vertebral strength decreased with increasing coefficient of variation of GLD, BV/TV, and E calculated from subregions of the vertebral centrum. The values of GLD, BV/TV, and E in centrum were significantly smaller for the anterior region than for the posterior region. Overall, these findings supported the significant role of regional variability of centrum properties in determining the whole vertebral strength.


Angle resolved Mueller polarimetry with a high numerical aperture and characterization of transparent biaxial samples.
We present a polarimetric instrument suitable for the simultaneous measurement of angle resolved normalized Mueller matrices for polar angles ranging from 0 degrees to 60 degrees and all azimuths. The polarimetric modulation and analysis are performed by means of an optimized polarization state generator and analyzer based on nematic liquid crystals. A high numerical aperture (0.95) microscope objective is used in double pass to illuminate the sample, with its rear focal plane imaged on a low noise CCD. This polarimeter can be used either in reflection, with the sample set in the objective front focal plane, or in transmission, for thin transparent samples. This latter configuration, which involves an additional spherical mirror with its center of curvature at the objective front focus, is described in detail. This instrument was used to accurately determine the directions of the optic axes and the angular dependence of the retardation of a biaxial polyethylene terephthalate (PET) plastic substrate in spite of the strong depolarization essentially due to the source 10 nm spectral width or the limitation in angular resolution due to the pixels distribution of the CCD combined with the sample large retardation.


Human and natural controls of the variation in aboveground tree biomass in African dry tropical forests.
Understanding the anthropogenic and natural controls that affect the patterns, distribution, and dynamics of terrestrial carbon is crucial to meeting climate change mitigation objectives. We assessed the human and natural controls over aboveground tree biomass density in African dry tropical forests, using Zambia's first nationwide forest inventory. We identified predictors that best explain the variation in biomass density, contrasted anthropogenic and natural sites at different spatial scales, and compared sites with different stand structure characteristics and species composition. In addition, we evaluated the effects of different management and conservation practices on biomass density. Variation in biomass density was mostly determined by biotic processes, linked with both species richness and dominance (evenness), and to a lesser extent, by land use, environmental controls, and spatial structure. Biomass density was negatively associated with tree species evenness and positively associated with species richness for both natural and human-modified sites. Human influence variables (including distance to roads, distance to town, fire occurrence, and the population on site) did not explain substantial variation in biomass density in comparison to biodiversity variables. The relationship of human activities to biomass density in managed sites appears to be mediated by effects on species diversity and stand structure characteristics, with lower values in human-modified sites for all metrics tested. Small contrasts in carbon density between human-modified and natural forest sites signal the potential to maintain carbon in the landscape inside but also outside forestlands in this region. Biodiversity is positively related to biomass density in both human and natural sites, demonstrating potential synergies between biodiversity conservation and climate change mitigation. This is the first evidence of positive outcomes of protected areas and participatory forest management on carbon storage at national scale in Zambia. This research shows that understanding controls over biomass density can provide policy relevant inputs for carbon management and on ecological processes affecting carbon storage.


Safety and pharmacokinetics of ganitumab (AMG 479) combined with sorafenib, panitumumab, erlotinib, or gemcitabine in patients with advanced solid tumors.
This phase 1b dose-escalation study assessed safety, tolerability, and pharmacokinetics of ganitumab, a fully human monoclonal antibody against the insulin-like growth factor 1 (IGF1) receptor, combined with targeted agents or cytotoxic chemotherapy in patients with advanced solid tumors. Patients with treatment-refractory advanced solid tumors were sequentially enrolled at 2 ganitumab dose levels (6 or 12 mg/kg i.v. every 2 weeks) combined with either sorafenib 400 mg twice daily, panitumumab 6 mg/kg every 2 weeks, erlotinib 150 mg once daily, or gemcitabine 1,000 mg/m(2) on days 1, 8, and 15 of each 4-week cycle. The primary end points were safety and pharmacokinetics of ganitumab. Ganitumab up to 12 mg/kg appeared well tolerated combined with sorafenib, panitumumab, erlotinib, or gemcitabine. Treatment-emergent adverse events were generally mild and included fatigue, nausea, vomiting, and chills. Three patients had dose-limiting toxicities: grade 3 hyperglycemia (ganitumab 6 mg/kg and panitumumab), grade 4 neutropenia (ganitumab 6 mg/kg and gemcitabine), and grade 4 thrombocytopenia (ganitumab 12 mg/kg and erlotinib). Ganitumab-binding and panitumumab-binding antibodies were detected in 5 and 2 patients, respectively; neutralizing antibodies were not detected. The pharmacokinetics of ganitumab and each cotherapy did not appear affected by coadministration. Circulating total IGF1 and IGF binding protein 3 increased from baseline following treatment. Four patients (9%) had partial responses. Ganitumab up to 12 mg/kg was well tolerated, without adverse effects on pharmacokinetics in combination with either sorafenib, panitumumab, erlotinib, or gemcitabine. Ganitumab is currently under investigation in combination with some of these and other agents.


Functional and electrophoretic characteristics of faba bean (Vicia faba) flour proteins as affected by germination.
Faba beans (Vicia faba) were germinated at room temperature for 3 and 6 days respectively. The effect of germination on the protein fractions, protein solubility index, PAGE pattern and some functional properties i.e. emulsification capacity (EC), foaming capacity (FC), foam stability (FS), water and fat absorption capacities of the flour was studied. Germination decreased albumins, globulins and prolamins at different levels but non protein nitrogen and glutelins were increased. The protein solubility index was high at both extreme pH values with an isoelectric point (IP) at pH of 4.4-4.5. The solubility of the protein slightly increased due to germination at all the pH values. PAGE pattern revealed on obvious dissociation and utilization for both fast and slow moving protein fractions during germination. Emulsification and foaming properties vs pH profile were similar to the pattern of solubility vs pH. Both properties were high at acidic and alkaline pH's and the minimum values were at pH 4 to 5. Germination process improved EC, FC and FS of the flour in comparison with that of dry bean flour. Water absorption of faba bean flours was improved during germination but the fat absorption markedly decreased.


Nanopore sequencing of a monkeypox virus strain isolated from a pustular lesion in the Central African Republic.
Monkeypox is an emerging and neglected zoonotic disease whose number of reported cases has been gradually increasing in Central Africa since 1980. This disease is caused by the monkeypox virus (MPXV), which belongs to the genus Orthopoxvirus in the family Poxviridae. Obtaining molecular data is particularly useful for establishing the relationships between the viral strains involved in outbreaks in countries affected by this disease. In this study, we evaluated the use of the MinION real-time sequencer as well as different polishing tools on MinION-sequenced genome for sequencing the MPXV genome originating from a pustular lesion in the context of an epidemic in a remote area of the Central African Republic. The reads corresponding to the MPXV genome were identified using two taxonomic classifiers, Kraken2 and Kaiju. Assembly of these reads led to a complete sequence of 196,956 bases, which is 6322 bases longer than the sequence previously obtained with Illumina sequencing from the same sample. The comparison of the two sequences showed mainly indels at the homopolymeric regions. However, the combined use of Canu with specific polishing tools such as Medaka and Homopolish was the best combination that reduced their numbers without adding mismatches. Although MinION sequencing is known to introduce a number of characteristic errors compared to Illumina sequencing, the new polishing tools allow a better-quality MinION-sequenced genome, thus to be used to help determine strain origin through phylogenetic analysis.


[Investigation of radioulnocarpal joint load distribution effects of radial shortening osteotomy for the treatment of Kienböck disease: a biomechanical study].
We evaluated the effects of radial shortening osteotomy on the radiolunate joint load distribution. This biomechanical study used standard left wrist models made of solid foam (Sawbones, Malmö, Sweden). The radioulnocarpal joint load distribution was investigated in the normal wrist model and 2 mm and 4 mm radial shortening osteotomy models under 14 kgf and 25 kgf loads when the wrist position was neutral. In the normal wrist group, the comparison of the average pressure distributions under the 14 kgf and 25 kgf loads shows that the increase at the ulnocarpal joint loading and the decrease at the radiolunate joint loading were statistically significant (p=0.012, p=0.036, respectively). When we compared the respective average pressure distributions after 14 kgf and 25 kgf loading, there were no significant differences between the normal wrist and the osteotomy groups. Consequently, radial shortening osteotomy is not effective in decreasing radiolunate joint loading when the wrist is in neutral position.


Achievable Rates for Pattern Recognition.
Biological and machine pattern recognition systems face a common challenge: Given sensory data about an unknown pattern, classify the pattern by searching for the best match within a library of representations stored in memory. In many cases, the number of patterns to be discriminated and the richness of the raw data force recognition systems to internally represent memory and sensory information in a compressed format. However, these representations must preserve enough information to accommodate the variability and complexity of the environment, otherwise recognition will be unreliable. Thus, there is an intrinsic tradeoff between the amount of resources devoted to data representation and the complexity of the environment in which a recognition system may reliably operate. In this paper, we describe a mathematical model for pattern recognition systems subject to resource constraints, and show how the aforementioned resource-complexity tradeoff can be characterized in terms of three rates related to the number of bits available for representing memory and sensory data, and the number of patterns populating a given statistical environment. We prove single-letter information-theoretic bounds governing the achievable rates, and investigate in detail two illustrative cases where the pattern data is either binary or Gaussian.


Impressions of sexual unfaithfulness and their accuracy show a degree of universality.
Forming accurate impressions of others' trustworthiness is a critical social skill, with faithfulness representing a key aspect of trust in sexual relationships. Interestingly, there is evidence for a small degree of accuracy in facial impressions of sexual unfaithfulness. Theoretical accounts suggest that these impressions may function to help with partner selection, and may be universal. If so, impressions should be similar for perceivers from different cultures and accuracy should not be limited to own-race faces. We tested these predictions by asking Caucasian and Asian women to judge the likelihood of unfaithfulness from the faces of Caucasian males whose past sexual history was known. In two studies we found high cross-cultural agreement in these impressions, consistent with universality in the impressions themselves. In Study 1, we found an other-race effect in impression accuracy, with significantly less accurate cross-race impressions by Asian women than own-race impressions by Caucasian women. Asian women showed no accuracy. Interestingly, in Study 2, Asian women who had grown up in the West showed small but significant accuracy in their impressions, with no other-race effect. Results are consistent with a degree of universality in the accuracy of this important aspect of social perception, provided that perceivers have experience with the faces being assessed.


Localizing the subunit pool for the temporally regulated polar pili of Caulobacter crescentus.
The pili of the stalked bacterium Caulobacter crescentus are assembled at a specific time in the life cycle at one pole of the cell and are composed of the monomer protein, pilin. A previous study demonstrated that the onset of pilin synthesis occurs well before pili appear on the surface, suggesting that pilin accumulates within the cell. In the present study, an electron microscope immunocytochemistry assay was used to determine the subcellular location of this unassembled pilin and its fate during pilus assembly and cell division. Populations of synchronously growing cells were embedded in epoxy resin at selected times during the cell cycle. Ultrathin sections were treated with pilin-specific antibody, followed by protein A coupled to colloidal gold. It was determined that the cellular location for unassembled pilin was the cell cytoplasm. All cell membranes and regions of nuclear material were poorly labeled. Quantitation demonstrated that label density increased during the period of pilin synthesis and declined during the period of pilus assembly and maintenance. The pilin pool was not unequally segregated at division; e.g., to the daughter cell that is elaborating pili. Mutants which have simultaneously lost the ability to produce flagella, pili, and other polar organelles, possibly due to alterations in the specialized region of polar organelle assembly, were also examined by the immunocytochemistry technique. There was no significant difference in the pilin pool size relative to the wild type, indicating that pilin synthesis continues in the absence of a functioning assembly site. This pattern of synthesis and assembly for the pilus is significantly different from that of the polar flagellum which is produced at the same time and location on the cell surface. These findings are discussed in relation to the hypothesized organization center at the cell pole which may have a major role in directing the assembly of all the polar structures.


Ultrasonographically predicting the extent of collateral flow through superficial temporal artery-to-middle cerebral artery anastomosis.
This study was performed to elucidate whether the extent of bypass flow through superficial temporal artery-to-middle cerebral artery (STA-MCA) anastomosis could be indirectly estimated by measuring the blood flow velocity in the superficial temporal artery (STA) by using duplex ultrasonography. We analyzed 29 patients (31 sides) who underwent STA-MCA bypass surgery for occlusive cerebrovascular disease (28 sides) or unclippable cerebral aneurysm that required therapeutic occlusion of the internal carotid artery (three sides). The flow velocities of the STA were measured by using ultrasonography. For patients who underwent the surgery unilaterally, the flow velocity ratios of the operated side to the contralateral side for the individual arteries were calculated. The correlation between these flow velocity parameters and the extent of bypass flow, which was graded based on the findings of cerebral angiography, was investigated. Both the affected STA flow velocity and the STA flow velocity ratio, particularly those in the end diastole, increased in patients with more extensive bypass flow. In patients with extensive, moderate, and poor bypass flow, the end diastolic flow velocities of the operated STA were 27.4 +/- 8.8, 23.0 +/- 7.8, and 13.5 +/- 7.5 cm/s, respectively and the end diastolic flow velocity ratios of the STA were 3.4 +/- 0.8, 2.1 +/- 0.5 and 1.3 +/- 0.4, respectively. The pulsatility index and resistance index of the affected STA were significantly lower in the patients with more extensive bypass flow. The optimal threshold value of the end diastolic flow velocity ratio of STA for the group with extensive bypass flow was 2.75, whereas that for the group with poor bypass flow was 1.60. With the obtained values, the sensitivity and specificity were 87.5% and 93.9% for the group with extensive bypass flow and 95.2% and 95.0% for the group with poor bypass flow, respectively. The blood flow velocity in the operated STA seems to be a highly sensitive parameter for predicting the extent of bypass flow in patients undergoing STA-MCA anastomosis.


Polyunsaturated fatty acids and recurrent mood disorders: Phenomenology, mechanisms, and clinical application.
A body of evidence has implicated dietary deficiency in omega-3 polyunsaturated fatty acids (n-3 PUFA), including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), in the pathophysiology and etiology of recurrent mood disorders including major depressive disorder (MDD) and bipolar disorder. Cross-national and cross-sectional evidence suggests that greater habitual intake of n-3 PUFA is associated with reduced risk for developing mood symptoms. Meta-analyses provide strong evidence that patients with mood disorders exhibit low blood n-3 PUFA levels which are associated with increased risk for the initial development of mood symptoms in response to inflammation. While the etiology of this n-3 PUFA deficit may be multifactorial, n-3 PUFA supplementation is sufficient to correct this deficit and may also have antidepressant effects. Rodent studies suggest that n-3 PUFA deficiency during perinatal development can recapitulate key neuropathological, neurochemical, and behavioral features associated with mood disorders. Clinical neuroimaging studies suggest that low n-3 PUFA biostatus is associated with abnormalities in cortical structure and function also observed in mood disorders. Collectively, these findings implicate dietary n-3 PUFA insufficiency, particularly during development, in the pathophysiology of mood dysregulation, and support implementation of routine screening for and treatment of n-3 PUFA deficiency in patients with mood disorders.


Germline progenitors escape the widespread phenomenon of homolog pairing during Drosophila development.
Homolog pairing, which plays a critical role in meiosis, poses a potential risk if it occurs in inappropriate tissues or between nonallelic sites, as it can lead to changes in gene expression, chromosome entanglements, and loss-of-heterozygosity due to mitotic recombination. This is particularly true in Drosophila, which supports organismal-wide pairing throughout development. Discovered over a century ago, such extensive pairing has led to the perception that germline pairing in the adult gonad is an extension of the pairing established during embryogenesis and, therefore, differs from the mechanism utilized in most species to initiate pairing specifically in the germline. Here, we show that, contrary to long-standing assumptions, Drosophila meiotic pairing in the gonad is not an extension of pairing established during embryogenesis. Instead, we find that homologous chromosomes are unpaired in primordial germ cells from the moment the germline can be distinguished from the soma in the embryo and remain unpaired even in the germline stem cells of the adult gonad. We further establish that pairing originates immediately after the stem cell stage. This pairing occurs well before the initiation of meiosis and, strikingly, continues through the several mitotic divisions preceding meiosis. These discoveries indicate that the spatial organization of the Drosophila genome differs between the germline and the soma from the earliest moments of development and thus argue that homolog pairing in the germline is an active process as versus a passive continuation of pairing established during embryogenesis.


ERK2 enters the nucleus by a carrier-independent mechanism.
In stimulated cells, the mitogen-activated protein kinase ERK2 (extracellular signal-regulated kinase 2) concentrates in the nucleus. Evidence exists for CRM1-dependent, mitogen-activated protein kinase kinase-mediated nuclear export of ERK2, but its mechanism of nuclear entry is not understood. To determine requirements for nuclear transport, we tagged ERK2 with green fluorescent protein (GFP) and examined its nuclear uptake by using an in vitro import assay. GFP-ERK2 entered the nucleus in a saturable, time- and temperature-dependent manner. Entry of GFP-ERK2, like that of ERK2, required neither energy nor transport factors and was visible within minutes. The nuclear uptake of GFP-ERK2 was inhibited by wheat germ agglutinin, which blocks nuclear entry by binding to carbohydrate moieties on nuclear pore complex proteins. The nuclear uptake of GFP-ERK2 also was reduced by excess amounts of recombinant transport factors. These findings suggest that ERK2 competes with transport factors for binding to nucleoporins, which mediate the entry and exit of transport factors. In support of this hypothesis, we showed that ERK2 binds directly to a purified nucleoporin. Our data suggest that GFP-ERK2 enters the nucleus by a saturable, facilitated mechanism, distinct from a carrier- and energy-dependent import mechanism and involves a direct interaction with nuclear pore complex proteins.


Nurses' perceptions of workplace culture in primary health care in Finland.
This study aimed to describe nurses' perceptions of workplace culture, especially in regard to stress levels, job satisfaction and the practice environment in primary health care. Health care is facing many challenges related to its attractiveness as a place of employment and the maintenance of a sufficient workforce supply. Previous studies report increasing rates of nurse job dissatisfaction and intentions to leave their current positions both in Finland and also globally. Improving workplace culture is thus vital in meeting the challenges related to recruitment and retention. A cross-sectional descriptive design was used to describe nurses' perceptions of workplace culture. Data were collected by questionnaire from 22 units in nine primary healthcare organizations in Finland, and analysed using descriptive and inferential statistics. Most of the respondents indicated that they were not certain whether their workplace culture was either positive or negative. Profession, age and work shift characteristics had an effect on the respondents' perceptions of workplace culture. Younger licensed practical and registered nurses assessed their workplace culture more positively, whereas older registered nurses and those working rotating rosters viewed workplace culture more negatively. The findings suggest that both unit and demographic characteristics affect workplace culture. This survey highlights that a positive workplace culture is one of the key factors in retaining and recruiting nurses, and provides an essential evidence that may be considered by other healthcare organizations. Nurse managers and healthcare leaders need to address workload management and take into account the related variables that affect a unit's workplace culture.


The impact of pedestrian countdown signals on pedestrian-motor vehicle collisions: a reanalysis of data from a quasi-experimental study.
To perform a more sophisticated analysis of previously published data that advances the understanding of the efficacy of pedestrian countdown signal (PCS) installation on pedestrian-motor vehicle collisions (PMVCs), in the city of Toronto, Canada. This is an updated analysis of the same dataset from Camden et al. A quasi-experimental design was used to evaluate the effect of PCS on PMVC. A Poisson regression analysis, using a one-group comparison of PMVC, pre-PCS installation to post-PCS installation was used, controlling for season and temporal effects. The outcome was the frequency of reported PMVC (January 2000-December 2009). Similar models were used to analyse specific types of collisions defined by age of pedestrian, injury severity, and pedestrian and vehicle action. Incidence rate ratios with 95% CI are presented. This analysis included 9262 PMVC, 2760 during or after PCS installation, at 1965 intersections. There was a 26% increase in the rate of collisions, pre to post-PCS installation (incidence rate ratio=1.26, 95% CI 1.11 to 1.42). The installation of PCS at 1965 signalised intersections in the city of Toronto resulted in an increase in PMVC rates post-PCS installation. PCSs may have an unintended consequence of increasing pedestrian-motor vehicle collisions in some settings.


Women's cancers: how the discovery of BRCA genes is driving current concepts of cancer biology and therapeutics.
Over the last two decades, discoveries related to the breast cancer susceptibility genes 1 and 2 (BRCA1 and BRCA2) have profoundly changed our understanding and management of hereditary breast and ovarian cancers. The concept of synthetic lethality, which arises when cells become vulnerable to a combination of deficiencies in DNA repair, has driven the expanding roles of poly (adenosine diphosphate (ADP)-ribose) polymerase inhibitors in breast and ovarian cancers, and prevention strategies are taking into account the tissue specificity, natural history (fallopian tube origin of some high-grade serous ovarian cancers) and hormone sensitivity of BRCA-associated cancers. Current research has focussed on further elucidating the roles of BRCA proteins in DNA repair, investigating other key DNA repair processes and proteins and linking aberrant DNA repair with carcinogenesis. The ultimate goal is to translate this evolving knowledge into improving the clinical care and treatment of patients with pathogenic BRCA variants or other deficiencies in homologous recombination (HR). In this review, we will discuss 1) the role of BRCA proteins in DNA repair; 2) emerging concepts in the biology of HR deficiency and 3) implications for prevention and treatment.


A single molecule investigation of the photostability of quantum dots.
Quantum dots (QDs) are very attractive probes for multi-color fluorescence imaging in biological applications because of their immense brightness and reported extended photostability. We report here however that single QDs, suitable for biological applications, that are subject to continuous blue excitation from a conventional 100 W mercury arc lamp will undergo a continuous blue-switching of the emission wavelength eventually reaching a permanent dark, photobleached state. We further show that β-mercaptoethanol has a dual stabilizing effect on the fluorescence emission of QDs: 1) by increasing the frequency of time that a QD is in its fluorescent state, and 2) by decreasing the photobleaching rate. The observed QD color spectral switching is especially detrimental for multi-color single molecule applications, as we regularly observe spectral blue-shifts of 50 nm, or more even after only ten seconds of illumination. However, of significant importance for biological applications, we find that even small, biologically compatible, concentrations (25 µM) of β-mercaptoethanol has a significant stabilizing effect on the emission color of QDs, but that greater amounts are required to completely abolish the spectral blue shifting or to minimize the emission intermittency of QDs.


Measuring time costs in interventions designed to reduce behavior problems among children and youth.
The economic evaluation of psychosocial interventions is a growing area of research. Though time costs are central to the economist's understanding of social costs, these costs generally have been ignored by prevention scientists. This article highlights the need to measure such costs and then reviews the principles economists use in valuing time. It then considers the specific time costs that often arise in interventions designed to reduce behavior problems among children and youth. These include classroom time devoted to program activities, the time of parents or other caregivers, the time of teachers (outside of the classroom), and the time of volunteers. We consider the economic principles that govern how economists value these inputs and then apply these principles to data from an evaluation of a prominent intervention in the field, the Incredible Years Program. We find that the time costs are potentially rather large and consider the implications for public policy of ignoring them.


An amorphous model for morphological processing in visual comprehension based on naive discriminative learning.
A 2-layer symbolic network model based on the equilibrium equations of the Rescorla-Wagner model (Danks, 2003) is proposed. The study first presents 2 experiments in Serbian, which reveal for sentential reading the inflectional paradigmatic effects previously observed by Milin, Filipović Đurđević, and Moscoso del Prado Martín (2009) for unprimed lexical decision. The empirical results are successfully modeled without having to assume separate representations for inflections or data structures such as inflectional paradigms. In the next step, the same naive discriminative learning approach is pitted against a wide range of effects documented in the morphological processing literature. Frequency effects for complex words as well as for phrases (Arnon & Snider, 2010) emerge in the model without the presence of whole-word or whole-phrase representations. Family size effects (Moscoso del Prado Martín, Bertram, Häikiö, Schreuder, & Baayen, 2004; Schreuder & Baayen, 1997) emerge in the simulations across simple words, derived words, and compounds, without derived words or compounds being represented as such. It is shown that for pseudo-derived words no special morpho-orthographic segmentation mechanism, as posited by Rastle, Davis, and New (2004), is required. The model also replicates the finding of Plag and Baayen (2009) that, on average, words with more productive affixes elicit longer response latencies; at the same time, it predicts that productive affixes afford faster response latencies for new words. English phrasal paradigmatic effects modulating isolated word reading are reported and modeled, showing that the paradigmatic effects characterizing Serbian case inflection have crosslinguistic scope.


An ultra-fast mechanically active cell culture substrate.
We present a mechanically active cell culture substrate that produces complex strain patterns and generates extremely high strain rates. The transparent miniaturized cell stretcher is compatible with live cell microscopy and provides a very compact and portable alternative to other systems. A cell monolayer is cultured on a dielectric elastomer actuator (DEA) made of a 30 μm thick silicone membrane sandwiched between stretchable electrodes. A potential difference of several kV's is applied across the electrodes to generate electrostatic forces and induce mechanical deformation of the silicone membrane. The DEA cell stretcher we present here applies up to 38% tensile and 12% compressive strain, while allowing real-time live cell imaging. It reaches the set strain in well under 1 ms and generates strain rates as high as 870 s-1, or 87%/ms. With the unique capability to stretch and compress cells, our ultra-fast device can reproduce the rich mechanical environment experienced by cells in normal physiological conditions, as well as in extreme conditions such as blunt force trauma. This new tool will help solving lingering questions in the field of mechanobiology, including the strain-rate dependence of axonal injury and the role of mechanics in actin stress fiber kinetics.


Salmonella serogroups identified from adult diarrhoeal out-patients in Addis Ababa, Ethiopia: antibiotic resistance and plasmid profile analysis.
Forty five Salmonella strains were isolated from 700 diarrhoeal samples collected from adult diarrhoeal out-patients in Addis Ababa. Among the isolates, serogroup C comprised 31.1%, B 24.4%, S.typhi 15.6%, D 13.3%, A 8.9% and E 6.7%. Among the isolates, 71.1% were resistant to tetracycline, 68.9% to ampicillin, 66.7% to cephalothin, 57.8% to trimethoprim-sulphamethoxazole, 53.9% to kanamycin, 46.7% to chloramphenicol and less than 31.8% of the isolates were resistant to other drugs. Among S.typhi isolates, 28.6% were resistant to chloramphenicol and this shows the emergence of chloramphenicol resistant S.typhi strains in Ethiopia. Gentamicin and polymyxin B were found to be the drugs of choice for cases of salmonellosis including S.typhi. All drug resistant isolates analysed for plasmids contained multiple plasmids ranging in sizes from 1.8 to greater than 21 Kilobases.


Respiratory chain complexes in dynamic mitochondria display a patchy distribution in life cells.
Mitochondria, the main suppliers of cellular energy, are dynamic organelles that fuse and divide frequently. Constraining these processes impairs mitochondrial is closely linked to certain neurodegenerative diseases. It is proposed that functional mitochondrial dynamics allows the exchange of compounds thereby providing a rescue mechanism. The question discussed in this paper is whether fusion and fission of mitochondria in different cell lines result in re-localization of respiratory chain (RC) complexes and of the ATP synthase. This was addressed by fusing cells containing mitochondria with respiratory complexes labelled with different fluorescent proteins and resolving their time dependent re-localization in living cells. We found a complete reshuffling of RC complexes throughout the entire chondriome in single HeLa cells within 2-3 h by organelle fusion and fission. Polykaryons of fused cells completely re-mixed their RC complexes in 10-24 h in a progressive way. In contrast to the recently described homogeneous mixing of matrix-targeted proteins or outer membrane proteins, the distribution of RC complexes and ATP synthase in fused hybrid mitochondria, however, was not homogeneous but patterned. Thus, complete equilibration of respiratory chain complexes as integral inner mitochondrial membrane complexes is a slow process compared with matrix proteins probably limited by complete fusion. In co-expressing cells, complex II is more homogenously distributed than complex I and V, resp. Indeed, this result argues for higher mobility and less integration in supercomplexes. Our results clearly demonstrate that mitochondrial fusion and fission dynamics favours the re-mixing of all RC complexes within the chondriome. This permanent mixing avoids a static situation with a fixed composition of RC complexes per mitochondrion.


Challenges in reproducing results from publicly available data: an example of sexual orientation and cardiovascular disease risk.
Replication is a vital part of the research process and has recently received considerable attention. Analyses using publicly available data should, if adequately described, be reproducible without assistance from the original investigators. Using data from the US National Health and Nutrition Examination Survey (NHANES), a recent study reported a statistically significant difference in cardiovascular disease risk comparing subgroups of sexual minority men. We attempted to reproduce these findings and assessed whether the results were robust to alternative analytic strategies and assumptions. We used the exclusion criteria and coding strategy described in the original paper to construct our analytical data set. Sampling weights were constructed in accordance with NHANES analytical guidelines. We estimated crude and covariate-adjusted associations between sexual orientation and vascular age using the regression models specified in the original report. We also conducted a series of sensitivity analyses to improve on the original findings. Our replication attempt was partially successful: we replicated the general trends reported in the original analysis, but not identical effect estimates. Importantly, we identified a potential misapplication of the Framingham Risk Score; correcting for this increased the probability that the reported null hypothesis test was a type I error. This paper supports the recent calls for greater transparency and improved reporting in research. Even with a publicly available and well-documented data source, we were unable to exactly replicate another study's original findings. Our sensitivity analyses revealed key issues in the original analysis and demonstrate the scientific importance of research replication.


Do ridge habitats contribute to pteridophyte diversity in tropical montane forests? A case study from southeastern Ecuador.
We address the question to which degree ridge habitats in tropical montane forests contribute to overall plant diversity by analysing patterns of pteridophyte (i.e. lycophytes and ferns) assemblages on ridges and slopes in three montane forest sites near Podocarpus National Park, Ecuador. The analyses, which involved 158 pteridophyte species (110 terrestrial, 96 epiphytic, 48 both) from 28 plots of 20 m x 20 m (or an equivalent of 400 m(2)), showed that more species were typical of one of the three study sites than of one of the two habitats (ridge/slope). As found in previous studies, alpha diversity on ridges was lower than on slopes, accounted for by the absence of numerous species that are found on slopes. Pteridophyte assemblages on ridges were more similar across study sites than those on slopes. Thus, unlike the structurally comparable (i.e. stunted, open) Amazonian forests, the studied montane ridge forests harbour fairly homogenous pteridophytes assemblages with very few specialised species. Our study implies that slope forests are of higher conservation priority for pteridophytes in the study region than ridge habitats. However, comparative studies are needed because other geographical regions and other groups of organisms may not share this pattern.


Optical trapping of metallic Rayleigh particles.
Metallic objects reflect light and have generally been considered poor candidates for optical traps, particularly with optical tweezers, which rely on a gradient force to provide trapping. We demonstrate that stable trapping can occur with optical tweezers when they are used with small metallic Rayleigh particles. In this size regime, the scattering pictures for metals and dielectrics are similar, and the larger polarizability of metals implies that trapping forces are greater. The latter fact makes the use of metal particles attractive for certain biological applications. Comparison of trapping forces for latex and gold spheres demonstrates that the gradient force is the major determinant of trapping strength and that competing effects, such as scattering or radiometric forces, are relatively minor.


A facile approach for synthesizing molecularly imprinted graphene for ultrasensitive and selective electrochemical detecting 4-nitrophenol.
In this work, a novel and convenient strategy was developed to prepare molecularly imprinted polymers (MIPs) on the surface of graphene sheet. In this route, vinyl group functionalized graphene (GR/NVC) was first prepared by immobilizing 4-vinylcarbazole onto the surface of graphene via π-π interaction. The subsequent grafting copolymerization of methacrylic acid and ethylene glycol dimethacrylate in the presence of 4-nitrophenol (4-NP, template molecule) was carried out at GR/NVC surface, leading to the formation of GR/MIPs composite. The GR/MIPs composite was characterized by FTIR, fluorescence, TGA, SEM and AFM, and was used to fabricate electrochemical sensor for the detection of 4-NP. The electrochemical behavior of GR/MIPs sensor for 4-NP was investigated by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The effects of the preparation conditions, such as concentration of the NVC and template, the solution pH, and incubation time, were also optimized. Under optimized conditions, the DPV current response of GR/MIPs sensor was nearly 12 times than that of the GR/NIPs sensor. It also should be noted that as compared to traditional MIP, shorter response time and much higher current response were demonstrated. In addition, the GR/MIPs sensor could recognize 4-NP from its structural analogs, indicating the excellent selectivity of the GR/MIPs sensor. The peak current is linearly proportional to the concentration of 4-NP ranging from 0.01 μM to 100 μM and 200 μM to 1000 μM with a significantly low detection limit of 5 nM, a wider response range and lower detection limits as compared to most of the previously reported electrochemical sensors for 4-NP. Furthermore, the GR/MIPs sensor exhibits good stability with adequate reproducibility and has been successfully used to determine 4-NP in water samples.


Differential effects of embryo transfer and maternal factors on anxiety-related behavior and numbers of neuropeptide Y (NPY) and parvalbumin (PARV) containing neurons in the amygdala of inbred C3H/HeN and DBA/2J mice.
The effects of reciprocal embryo transfers were studied on anxiety-related behavior of inbred C3H/HeN and DBA/2J mice on the elevated plus maze (EPM), and related to amygdaloid neuropeptide Y (NPY)- and parvalbumin (PARV)-immunoreactive neurons. Embryo transfer significantly reduced closed arm entries in in-stain-transferred C3H mice, and maternal factors influenced open arm entries only in interaction with genetic background and sex. In DBA/2J-mice, embryo transfer resulted in a reduced number of NPY-immunoreactive (NPY-ir) neurons, while PARV-immunoreactive (PARV-ir) cells were not affected. In C3H/HeN mice, however, in-strain embryo transfer only resulted in a reduction of the number of PARV-immunoreactive neurons. Maternal factors mainly induced changes in the number of NPY-ir neurons in the basolateral complex of the amygdala either directly or in interaction with genetic factors. In summary, in-strain embryo transfer had a minor effect on the behavior of C3H/HeN mice, and a differential influence on the numbers of amygdaloid NPY-ir and PARV-ir neurons of inbred C3H/HeN and DBA/2J mice. Maternal factors had a stronger impact on the numbers of NPY-ir neurons than PARV-ir neurons. The present results indicate that alterations in behavior and amygdala morphology induced by embryo transfer or maternal factors depend on the genetic background of the mouse strains used.


Cardiovascular disease event rates in patients with severe psoriasis treated with systemic anti-inflammatory drugs: a Danish real-world cohort study.
Psoriasis is a chronic inflammatory disorder associated with cardiovascular morbidity and mortality. Systemic anti-inflammatory drugs, including biological agents, are widely used in the treatment of patients with moderate to severe psoriasis and may attenuate the risk of cardiovascular disease events. We therefore examined the rate of cardiovascular disease events in patients with severe psoriasis treated with systemic anti-inflammatory drugs. Individual-level linkage of nationwide administrative databases was used to assess the event rates associated with use of biological agents, methotrexate or other therapies, including retinoids, cyclosporine and phototherapy, in Denmark from 2007 to 2009. Death, myocardial infarction and stroke. A total of 2400 patients with severe psoriasis, including 693 patients treated with biological agents and 799 treated with methotrexate, were identified. Incidence rates per 1000 patient-years and 95% confidence intervals (CIs) for the composite endpoint were 6.0 (95% CI 2.7-13.4), 17.3 (95% CI 12.3-24.3) and 44.5 (95% CI 34.6-57.0) for patients treated with biological agents, methotrexate and other therapies, respectively. Age- and sex-adjusted hazard ratios (HRs) were 0.28 (95% CI 0.12-0.64) and 0.65 (95% CI 0.42-1.00) for patients treated with biological agents and methotrexate, respectively, using other therapies as the reference cohort. Corresponding HRs for a secondary composite endpoint of cardiovascular death, myocardial infarction and stroke were 0.48 (95% CI 0.17-1.38) and 0.50 (95% CI 0.26-0.97). In this nationwide study of patients with severe psoriasis, systemic anti-inflammatory treatment with biological agents or methotrexate was associated with lower cardiovascular disease event rates compared to patients treated with other anti-psoriatic therapies.


Complement Factor H-Related 5-Hybrid Proteins Anchor Properdin and Activate Complement at Self-Surfaces.
C3 glomerulopathy (C3G) is a severe kidney disease for which no specific therapy exists. The causes of C3G are heterogeneous, and defective complement regulation is often linked to C3G pathogenesis. Copy number variations in the complement factor H-related (CFHR) gene cluster on chromosome 1q32 and CFHR5 mutant proteins associate with this disease. Here, we identified CFHR5 as a pattern recognition protein that binds to damaged human endothelial cell surfaces and to properdin, the human complement activator. We found the two N-terminal short consensus repeat domains of CFHR5 contact properdin and mediate dimer formation. These properdin-binding segments are duplicated in two mutant CFHR5 proteins, CFHR2-CFHR5Hyb from German patients with C3G and CFHR5Dup from Cypriot patients with C3G. Each of these mutated proteins assembled into large multimeric complexes and, compared to CFHR5, bound damaged human cell surfaces and properdin with greater intensity and exacerbated local complement activation. This enhanced surface binding and properdin recruitment was further evidenced in the mesangia of a transplanted and explanted kidney from a German patient with a CFHR2-CFHR5Hyb protein. Enhanced properdin staining correlated with local complement activation with C3b and C5b-9 deposition on the mesangial cell surface in vitro This gain of function in complement activation for two disease-associated CFHR5 mutants describes a new disease mechanism of C3G, which is relevant for defining appropriate treatment options for this disorder.


Trackways Produced by Lungfish During Terrestrial Locomotion.
Some primarily aquatic vertebrates make brief forays onto land, creating traces as they do. A lack of studies on aquatic trackmakers raises the possibility that such traces may be ignored or misidentified in the fossil record. Several terrestrial Actinopterygian and Sarcopterygian species have previously been proposed as possible models for ancestral tetrapod locomotion, despite extant fishes being quite distinct from Devonian fishes, both morphologically and phylogenetically. Although locomotion has been well-studied in some of these taxa, trackway production has not. We recorded terrestrial locomotion of a 35 cm African lungfish (Protopterus annectens; Dipnoi: Sarcopterygii) on compliant sediment. Terrestrial movement in the lungfish is accomplished by planting the head and then pivoting the trunk. Impressions are formed where the head impacts the substrate, while the body and fins produce few traces. The head leaves a series of alternating left-right impressions, where each impact can appear as two separate semi-circular impressions created by the upper and lower jaws, bearing some similarity to fossil traces interpreted as footprints. Further studies of trackways of extant terrestrial fishes are necessary to understand the behavioural repertoire that may be represented in the fossil track record.


Regeneration of peri-implant infrabony defects using PerioGlas: a pilot study in rabbits.
PerioGlas is a silicate-based synthetic bone augmentation material that has been used to fill periodontal defects with bonding and integration to both soft tissue and bone. The purpose of this research was to determine the PerioGlas interface with titanium dental implants and bone. Seven live rabbits were used; however, one rabbit was euthanized at 3 days as a result of a tibial fracture through the implant placement site. Each rabbit received four 3.3 x 8 mm Imtec titanium plasma-sprayed dental implants, two in each proximal tibia. One implant in each rabbit was placed in the standard fashion. Two implants in each rabbit had a surgically created defect adjacent to one side of the coronal aspect of the implant. The defect was subsequently filled with PerioGlas. One implant in each rabbit had a surgically created defect that was not filled with PerioGlas. The rabbits were sacrificed at 1, 2, 3, 6, 12, and 24 weeks. Each specimen was prepared for histologic viewing, yielding a nondecalcified specimen demonstrating the interface of bone, implant, and PerioGlas. The results demonstrate peripheral formation of osteoid, followed by bone deposition within the defect from host (surgical margin) bone, toward the implant. The new osteoid and bone form around the PerioGlas particles. Newly formed trabeculae connect these areas of osteoid and new bone around the PerioGlas, interconnecting the PerioGlas particles. The new bone eventually reaches the implant, and osseointegration occurs with incorporation of the PerioGlas particles.


[Mental patients and their profile of compliance with psychopharmacological treatment].
Non-compliance with drug-based treatment is a complex and universal phenomenon that develops gradually during psychiatric treatments, and is related to disease aggravation. The purpose of this study was to describe the profile of using a given medication in the clientele of a community mental healthcare service and its characteristics. We performed a retrospective study in the medical records of patients who used Haloperidol Decanoate in a period of 1 year and 8 months. 167 medical records were analyzed. 60% of the patients presented irregular use of medication, which reflects in high risks of relapse and re-hospitalizations. This profile configures low compliance with the prescribed drug treatment, leading to reflections about which factors influence this behavior and which measures could be implemented in the maintenance of extra-hospital treatment for these patients.


Fever in geriatric emergency patients: clinical features associated with serious illness.
To determine the clinical significance of fever in geriatric emergency department patients. Case series with follow-up. Urban, university-affiliated community hospital. Consecutive patients over the age of 65 years who presented to the ED during a 12-month period with an oral temperature of 100.0 degrees F (37.8 degrees C) or higher. We considered the following features indicators of serious illness: positive blood culture(s), related death within 1 month of ED visit, need for surgery or other invasive procedure, hospitalization for 4 or more days, IV antibiotics for 3 or more days, and repeat ED visit within 72 hours for related condition. Four hundred eighty-nine patients were eligible for study. Of the 470 patients with complete follow-up data, 357 (76.0%) had indicators of serious illness. Clinical features found to be independently associated with serious illness included oral temperature of 103 degrees F (39.4 degrees C) or more, respiration rate of 30 or more, leukocytosis of 11.0 x 10(9)/L or more, presence of an infiltrate, and pulse of 120 or more. At least one indicator of serious illness was present in 63 of 128 patients (49.6%) with none of these independently predictive clinical features. The most common final diagnoses were pneumonia (24.0%), urinary-tract infection (21.7%), and sepsis (12.8%). Fever among geriatric ED patients frequently marks the presence of serious illness. All such patients should be strongly considered for hospital admission, particularly when certain clinical features are present. The absence of abnormal findings does not reliably rule out the possibility of serious illness.


Diosmin ameliorative effects on oxidative stress and fibrosis in paraquat-induced lung injury in mice.
Paraquat (PQ) induces pulmonary fibrosis, a progressive lung disorder resulting in severe respiratory failure and death. Increased oxidative stress, inflammatory reactions, and multiple fibrotic lesions are major features of PQ-induced lung injury. Diosmin (Dio) is a safe drug that is available for clinical use for vascular disorders. Dio exhibits antioxidant, anti-inflammatory, and antifibrotic activities. Accordingly, the aim of this study was to evaluate the protective effect of diosmin on PQ-induced lung injury in mice and the underlying mechanisms involved. Lung injury was induced by PQ (30 mg/kg, intraperitoneally) in NMRI albino mice and Dio (50 and 100 mg/kg, gavage) was administrated 3 days before PQ and continued for 10 or 24 days. After euthanizing the mice, the biochemical and histopathological markers of lung tissue were determined. PQ significantly increased oxidative stress, inflammatory, and fibrotic markers. PQ increased the level of malonedaldehyde (MDA) and hydroxyproline (HYP) and decreased the level of glutathione (GSH) and catalase activity in the lung. Dio (50 and 100 mg/kg) significantly increased GSH levels and catalase activity and decreased HYP content and MDA levels. In addition, Dio reduced histopathological injuries in hematoxylin and eosin-stained and Masson's trichrome-stained sections. These findings suggest that Dio has protective effects against PQ-induced lung injury, which may be due to its antioxidant, anti-inflammatory, and antifibrotic effects.


Dinosaur body temperatures determined from isotopic (¹³C-¹⁸O) ordering in fossil biominerals.
The nature of the physiology and thermal regulation of the nonavian dinosaurs is the subject of debate. Previously, arguments have been made for both endothermic and ectothermic metabolisms on the basis of differing methodologies. We used clumped isotope thermometry to determine body temperatures from the fossilized teeth of large Jurassic sauropods. Our data indicate body temperatures of 36° to 38°C, which are similar to those of most modern mammals. This temperature range is 4° to 7°C lower than predicted by a model that showed scaling of dinosaur body temperature with mass, which could indicate that sauropods had mechanisms to prevent excessively high body temperatures being reached because of their gigantic size.


E-readers are more effective than paper for some with dyslexia.
E-readers are fast rivaling print as a dominant method for reading. Because they offer accessibility options that are impossible in print, they are potentially beneficial for those with impairments, such as dyslexia. Yet, little is known about how the use of these devices influences reading in those who struggle. Here, we observe reading comprehension and speed in 103 high school students with dyslexia. Reading on paper was compared with reading on a small handheld e-reader device, formatted to display few words per line. We found that use of the device significantly improved speed and comprehension, when compared with traditional presentations on paper for specific subsets of these individuals: Those who struggled most with phoneme decoding or efficient sight word reading read more rapidly using the device, and those with limited VA Spans gained in comprehension. Prior eye tracking studies demonstrated that short lines facilitate reading in dyslexia, suggesting that it is the use of short lines (and not the device per se) that leads to the observed benefits. We propose that these findings may be understood as a consequence of visual attention deficits, in some with dyslexia, that make it difficult to allocate attention to uncrowded text near fixation, as the gaze advances during reading. Short lines ameliorate this by guiding attention to the uncrowded span.


Dietary fiber and progression of atherosclerosis: the Los Angeles Atherosclerosis Study.
Several epidemiologic studies found weak protective relations between dietary fiber intake and the risk of cardiovascular disease events. However, few of the studies addressed possible mechanisms of the effect. In the present study, we estimated relations between the progression of atherosclerosis and the intake of selective dietary fiber fractions. Mediation of the relations by serum lipids was also investigated. Participants who were free of heart disease and aged 40-60 y were recruited into the cohort (n = 573; 47% women). The intima-media thickness (IMT) of the common carotid arteries was measured ultrasonographically at the baseline examination and at 2 follow-up examinations (n = 500), dietary intakes were assessed with six 24-h recalls (3 at baseline and 3 at the first follow-up examination), and blood samples were analyzed at baseline and at both follow-up examinations. A significant inverse association was observed between IMT progression and the intakes of viscous fiber (P = 0.05) and pectin (P = 0.01). Correction for measurement error increased the magnitude of these estimated effects. The ratio of total to HDL cholesterol was inversely related to the intakes of total fiber (P = 0.01), viscous fiber (P = 0.05), and pectin (P = 0.01). The magnitude of the association between IMT progression and the intakes of viscous fiber and pectin was attenuated by adjustment for serum lipids. The intake of viscous fiber, especially pectin, appears to protect against IMT progression. Serum lipids may act as a mediator between dietary fiber intake and IMT progression.


Psychological interventions for bipolar disorder in low- and middle-income countries: systematic review.
Adjunctive psychological interventions for bipolar disorder have demonstrated better efficacy in preventing or delaying relapse and improving outcomes compared with pharmacotherapy alone. To evaluate the efficacy of psychological interventions for bipolar disorder in low- and middle-income countries. A systematic review was conducted using PubMed, PsycINFO, Medline, EMBASE, Cochrane database for systematic review, Cochrane central register of controlled trials, Latin America and Caribbean Center on Health Science Literature and African Journals Online databases with no restriction of language or year of publication. Methodological heterogeneity of studies precluded meta-analysis. A total of 18 adjunctive studies were identified: psychoeducation (n = 14), family intervention (n = 1), group cognitive-behavioural therapy (CBT) (n = 2) and group mindfulness-based cognitive therapy (MBCT) (n = 1). In total, 16 of the 18 studies were from upper-middle-income countries and none from low-income countries. All used mental health specialists or experienced therapists to deliver the intervention. Most of the studies have moderately high risk of bias. Psychoeducation improved treatment adherence, knowledge of and attitudes towards bipolar disorder and quality of life, and led to decreased relapse rates and hospital admissions. Family psychoeducation prevented relapse, decreased hospital admissions and improved medication adherence. CBT reduced both depressive and manic symptoms. MBCT reduced emotional dysregulation. Adjunctive psychological interventions alongside pharmacotherapy appear to improve the clinical outcome and quality of life of people with bipolar disorder in middle-income countries. Further studies are required to investigate contextual adaptation and the role of non-specialists in the provision of psychological interventions to ensure scalability and the efficacy of these interventions in low-income country settings. None.


The influence of organic production on food quality - research findings, gaps and future challenges.
Although several meta-analysis studies have been published comparing the quality of food derived from organic and non-organic origin, it is still not clear if food from organic production per se can guarantee product-related added value to consumers. This paper aims to summarize the status quo in order to identify research gaps and suggest future research challenges. Organic food is described according to a quality model already published. The influence of organic production on food quality is structured in primary production and processing. Furthermore, organic food authentication is discussed. Organic food seems to contain fewer pesticide residues and statistically more selected health-related compounds such as polyphenols in plant products and polyunsaturated fatty acids in milk and meat products, but the health relevance for consumers is not clear yet. Comparing food from organic origin with so called 'conventional' food seems not to be appropriate, because 'conventional' is not defined. In organic food quality research a system approach is needed from which systemic markers can be selected. Research on the impact of processing technologies on the quality according to organic principles seems of high relevance, since most of the food is processed.


Effect of nanosecond laser pre-irradiation on the femtosecond laser-induced damage of Ta2O5/SiO2 high reflector.
The effect of nanosecond laser pre-irradiation on the femtosecond laser-induced damage behaviors of 800 nm 0° AOI Ta(2)O(5)/SiO(2) high reflectors fabricated by e-beam evaporation was explored. Laser pre-irradiation was carried out by Raster-scanning with scanning mode of 1-on-1 and scanning velocities timed such that there was a beam overlap at 70% of the peak fluence, utilizing 5 Hz 1064 nm 12 ns Nd:YAG fundamental lasers. Femtosecond laser damage was investigated by 1 kHz 800 nm 135 fs Ti: sapphire laser system with 1-on-1 mode test. The results indicated that nanosecond laser pre-irradiation did not promote the femtosecond laser-induced damage threshold of reflectors. Instead, the thresholds of all the samples with various fluence steps for pre-irradiation were reduced by about 20%. Furthermore, the damage morphologies were analyzed by optical microscope, SEM and AFM, which displayed deterministic field induced breakdown characteristics. To explain these phenomena, a theoretical model including photoionization, avalanche ionization, and decays of electrons was built to simulate the evolution of electron density in the conduction band. Field ionization mechanism was considered to dominate the femtosecond laser damage process, while the electronic defects induced by nanosecond laser pre-irradiation accelerated the femtosecond laser damage evolution.


Gene expression endpoints following chronic waterborne copper exposure in a genomic model organism, the zebrafish, Danio rerio.
Although copper (Cu) is an essential micronutrient for all organisms, in excess, waterborne Cu poses a significant threat to fish from the cellular to population level. We examined the physiological and gene expression endpoints that chronic waterborne Cu exposure (21 d) imposes on soft-water acclimated zebrafish at two environmentally relevant concentrations: 8 microg/l (moderate) and 15 microg/l (high). Using a 16,730 65-mer oligonucleotide customized zebrafish microarray chip related to metal metabolism and toxicity to assess the transcriptomic response, we found that 573 genes in the liver responded significantly to Cu exposure. These clustered into three distinct patterns of expression. There was distinct upregulation of a majority of these genes under moderate Cu exposure and a significant downregulation under high Cu exposure. Microarray results were validated by qPCR of eight genes; two genes, metallothionein 2 (mt2) and Na(+)-K(+)-ATPase 1a1 (atp1a1), displayed increased expression under both Cu exposures, indicative of potential genetic endpoints of Cu toxicity, whereas the remaining six genes demonstrated opposing effects at each Cu exposure. Na(+)-K(+)-ATPase enzyme activity decreased during Cu exposure, which may be linked to Cu's competitive effects with Na(+). Whole body cortisol levels were significantly increased in Cu-exposed fish, which prompted an analysis of the promoter region of all significantly regulated genes for glucocorticoid (GRE) and metal (MRE) response elements to dissociate metal- and stress-specific gene responses. Of the genes significantly regulated, 30% contained only a GRE sequence, whereas 2.5% contained only a consensus MRE. We conclude that the indirect effects of Cu exposure regulate gene expression to a much greater degree than the direct effects.


Music and the heart.
Music can powerfully evoke and modulate emotions and moods, along with changes in heart activity, blood pressure (BP), and breathing. Although there is great heterogeneity in methods and quality among previous studies on effects of music on the heart, the following findings emerge from the literature: Heart rate (HR) and respiratory rate (RR) are higher in response to exciting music compared with tranquilizing music. During musical frissons (involving shivers and piloerection), both HR and RR increase. Moreover, HR and RR tend to increase in response to music compared with silence, and HR appears to decrease in response to unpleasant music compared with pleasant music. We found no studies that would provide evidence for entrainment of HR to musical beats. Corresponding to the increase in HR, listening to exciting music (compared with tranquilizing music) is associated with a reduction of heart rate variability (HRV), including reductions of both low-frequency and high-frequency power of the HRV. Recent findings also suggest effects of music-evoked emotions on regional activity of the heart, as reflected in electrocardiogram amplitude patterns. In patients with heart disease (similar to other patient groups), music can reduce pain and anxiety, associated with lower HR and lower BP. In general, effects of music on the heart are small, and there is great inhomogeneity among studies with regard to methods, findings, and quality. Therefore, there is urgent need for systematic high-quality research on the effects of music on the heart, and on the beneficial effects of music in clinical settings.


Induction of cytochrome P450 family 1 mRNAs and activities in a cell line from the frog Xenopus laevis.
Cytochrome P450 family 1 (CYP1) includes four subfamilies of enzymes: CYP1A, CYP1B, CYP1C, and CYP1D. In many vertebrates, CYP1A, 1B, and 1C expression is induced by agonists of the aryl hydrocarbon receptor, including toxic contaminants such as chlorinated dioxins, coplanar chlorinated biphenyls, and polynuclear aromatic hydrocarbons. Assessed at the level of mRNA, protein, or enzyme activity, CYP1s (especially CYP1As) represent potent and popular biomarkers of contaminant exposure in aquatic vertebrates. Alkylated resorufins are synthetic substrates used to detect, quantify, and describe catalytic activities of cytochrome P450s. The ability to oxidize specific resorufin-based substrates can distinguish the catalytic activities of individual CYP1s. Xenopus laevis, the African clawed frog, is the most widely employed amphibian model in aquatic toxicology, yet the number, inducibility, and activities of CYP1s have not been systematically characterized in this species. Here we report the cloning of cDNAs encoding two new CYP1 family members, X. laevis CYP1B and CYP1C, along with an integrated assessment of the induction of alkyloxyuresorufin-O-dealkylase (AROD) activities and mRNA expression of four known X. laevis CYP1s: CYP1A6, CYP1A7, CYP1B, and CYP1C. Using XLK-WG, an X. laevis kidney epithelial cell line, we determined that EROD (ethoxyresorufin substrate) and MROD (methoxyresorufin) were both induced 3000- to 5000-fold following 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) exposure up to 250 nM, while BROD (benzyloxyresorufin) and PROD (pentyloxyresorufin) activity was not detectable regardless of TCDD treatment. TCDD induced CYP1A6 and CYP1A7 mRNAs by 2-3 orders of magnitude, while CYP1B and CYP1C were unchanged. The more potent AHR agonist, FICZ (6-formylindolo[3,2-b]carbazole), induced CYP1B up to 10-fold at concentrations between 0.1 and 250 nM, while CYP1C induction was less than 3-fold. CYP1B mRNA showed the highest constitutive mRNA expression, 5- to 75-fold greater than the other CYP1 transcripts. Taken together, these results suggest that CYP1A6 and CYP1A7 perform the bulk of EROD and MROD activities we observed in these cells. The ability of each X. laevis CYP1 to catalyze oxidation of individual resorufin substrates remains to be determined. Correlating CYP1 mRNA and induced AROD activity is a significant step toward clarifying the biochemical meaning of these biomarkers and the roles of CYP1 enzymes in X. laevis. The cell culture approach represents an important complement to the long standing use of frog embryos and tadpoles in toxicological studies, providing a well suited model system for determining the molecular mechanisms underlying the regulation of these important biomarkers of contaminant exposure.


Itch/AIP4 mediates Deltex degradation through the formation of K29-linked polyubiquitin chains.
Deltex (DTX) and AIP4 are the human orthologues of the Drosophila deltex and Suppressor of deltex, which have been genetically described as being antagonistically involved in the Notch signalling pathway. Both genes encode E3 ubiquitin ligases of the RING (Really interesting new gene)-H2 and HECT (Homologous to E6AP carboxyl terminus) families, respectively. In an attempt to understand the molecular basis of their genetic interactions, we studied the relationship between DTX and AIP4 in the absence of activation of the Notch pathway. We show here that both molecules interact and partially colocalize to endocytic vesicles, and that AIP4 targets DTX for lysosomal degradation. Furthermore, AIP4-generated polyubiquitin chains are mainly conjugated through lysine 29 of ubiquitin in vivo, indicating a link between this type of chain and lysosomal degradation.


Prenatal diagnosis of Bartter syndrome: amniotic fluid aldosterone.
Bartter syndrome is a severe inherited tubulopathy characterized at birth by salt wasting, severe polyuria, dehydration, growth retardation and secondary hyperaldosteronism. Prenatally, the disease is usually discovered following onset of severe polyhydramnios. We studied amniotic fluid aldosterone concentration in cases of Bartter syndrome and in control groups. Amniotic fluid aldosterone was assayed by radioimmunoassay. We undertook a retrospective case-control study based on 36 cases of postnatally diagnosed Bartter syndrome and 144 controls matched for gestational age. Two controls groups were defined: controls with polyhydramnios (n=72) and control without polyhydramnios (n=72). Amniotic fluid aldosterone was compared between the three groups. The median amniotic fluid aldosterone concentration in the Bartter syndrome group (90 pg/mL) did not differ significantly from that in the controls with polyhydramnios (90 pg/mL, p=0.33) or the controls without polyhydramnios (87 pg/mL, p=0.41). In conclusion, amniotic fluid aldosterone assay cannot be used for prenatal diagnosis of Bartter syndrome.