High-throughput sequencing approach in analysis of microbial communities colonizing natural gas pipelines.
This study provides a deep modern insight into the phylogenetic diversity among bacterial consortia found in working and nonworking high-methane natural gas pipelines located in Poland. The working pipeline was characterized by lower biodiversity (140-154 bacterial genera from 22 to 23 classes, depending on the source of the debris) in comparison to the off-gas pipeline (169 bacterial genera from 23 classes). The sediment recovered from the working pipeline contained mostly DNA identified as belonging to the phylum Firmicutes (66.4%-45.9% operational taxonomic units [OTUs]), predominantly Bacillus (41.4%-31.1% OTUs) followed by Lysinibacillus (2.6%-1.5% OTUs) and Clostridium (2.4%-1.8% OTUs). In the nonworking pipeline, Proteobacteria (46.8% OTUs) and Cyanobacteria (27.8% OTUs) were dominant. Over 30% of the Proteobacteria sequences showed homologies to Gammaproteobacteria, with Pseudomonas (7.1%), Enhydrobacter (2.1%), Stenotrophomonas (0.5%), and Haempohilus (0.4%) among the others. Differences were noted in terms of the chemical compositions of deposits originating from the working and nonworking gas pipelines. The deposits from the nonworking gas pipeline contained iron, as well as carbon (42.58%), sulphur (15.27%), and oxygen (15.32%). This composition can be linked to both the quantity and type of the resident microorganisms. The presence of a considerable amount of silicon (17.42%), and of aluminum, potassium, calcium, and magnesium at detectable levels, may likewise affect the metabolic activity of the resident consortia in the working gas pipeline. All the analyzed sediments included both bacteria known for causing and intensifying corrosion (e.g., Pseudomonas, Desulfovibrio, Shewanella, Serratia) and bacteria that can protect the surface of pipelines against deterioration (e.g., Bacillus). Biocorrosion is not related to a single mechanism or one species of microorganism, but results from the multidirectional activity of multiple microbial communities. The analysis presented here of the state of the microbiome in a gas pipeline during the real gas transport is a particularly valuable element of this work.


Academic Institutions' Critical Guidelines for Health Care Workers Who Deploy to West Africa for the Ebola Response and Future Crises.
The unprecedented Ebola Virus Disease (EVD) outbreak in West Africa, with its first cases documented in March 2014, has claimed the lives of thousands of people, and it has devastated the health care infrastructure and workforce in affected countries. Throughout this outbreak, there has been a critical lack of health care workers (HCW), including physicians, nurses, and other essential non-clinical staff, who have been needed, in most of the affected countries, to support the medical response to EVD, to attend to the health care needs of the population overall, and to be trained effectively in infection protection and control. This lack of sufficient and qualified HCW is due in large part to three factors: 1) limited HCW staff prior to the outbreak, 2) disproportionate illness and death among HCWs caused by EVD directly, and 3) valid concerns about personal safety among international HCWs who are considering responding to the affected areas. These guidelines are meant to inform institutions who deploy professional HCWs.


Inverse size scaling of the nucleolus by a concentration-dependent phase transition.
Just as organ size typically increases with body size, the size of intracellular structures changes as cells grow and divide. Indeed, many organelles, such as the nucleus [1, 2], mitochondria [3], mitotic spindle [4, 5], and centrosome [6], exhibit size scaling, a phenomenon in which organelle size depends linearly on cell size. However, the mechanisms of organelle size scaling remain unclear. Here, we show that the size of the nucleolus, a membraneless organelle important for cell-size homeostasis [7], is coupled to cell size by an intracellular phase transition. We find that nucleolar size directly scales with cell size in early C. elegans embryos. Surprisingly, however, when embryo size is altered, we observe inverse scaling: nucleolar size increases in small cells and decreases in large cells. We demonstrate that this seemingly contradictory result arises from maternal loading of a fixed number rather than a fixed concentration of nucleolar components, which condense into nucleoli only above a threshold concentration. Our results suggest that the physics of phase transitions can dictate whether an organelle assembles, and, if so, its size, providing a mechanistic link between organelle assembly and cell size. Since the nucleolus is known to play a key role in cell growth, this biophysical readout of cell size could provide a novel feedback mechanism for growth control.


Pairs of pigeons act as behavioural units during route learning and co-navigational leadership conflicts.
In many species, group members obtain benefits from moving collectively, such as enhanced foraging efficiency or increased predator detection. In situations where the group's decision involves integrating individual preferences, group cohesion can lead to more accurate outcomes than solitary decisions. In homing pigeons, a classic model in avian orientation studies, individuals learn habitual routes home, but whether and how co-navigating birds acquire and share route-based information is unknown. Using miniature GPS loggers, we examined these questions by first training pairs (the smallest possible flocks) of pigeons together, and then releasing them with other pairs that had received separate pair-training. Our results show that, much like solitary individuals, pairs of birds are able to establish idiosyncratic routes that they recapitulate together faithfully. Also, when homing with other pairs they exhibit a transition from a compromise- to a leadership-like mechanism of conflict resolution as a function of the degree of disagreement (distance separating the two preferred routes) between the two pairs, although pairs tolerate a greater range of disagreements prior to the transition than do single birds. We conclude that through shared experiences during past decision-making, pairs of individuals can become units so closely coordinated that their behaviour resembles that of single birds. This has implications for the behaviour of larger groups, within which certain individuals have closer social affiliations or share a history of previous associations.


MO-F-213AB-02: Correcting Spread-Out Bragg Peak Slope Using Time-Resolved Monte Carlo Simulations and Beam Current Modulation.
To fully characterize the spread-out Bragg peak from the Mevion S250 Proton Therapy System using Monte Carlo simulations, evaluate the slope of the spread-out Bragg peak in the treatment field, and correct this slope by virtually applying beam current modulation. MCNPX simulations were performed on the Mevion S250 for each of the 24 beamline configurations, with the modulator wheel rotated in 1 degree increments. Energy deposition was tallied in a water phantom, resulting in 8,640 central axis depth-dose calculations. These data were imported into MATLAB and the slope of the spread-out Bragg peak (SOBP) was evaluated for a constant beam current. An iterative algorithm was developed to determine the optimal beam current modulation (BCM) profile as a function of modulator wheel position for each configuration. These BCM profiles were then applied virtually to the MCNPX SOBP data and slope values were recalculated for the optimized SOBP profiles. Ideally SOBP slope would measure 0.0 %/cm in the treatment field, corresponding to a uniform dose delivery. Prior to BCM correction, slopes between -1.0 and +2.0 %/cm were observed across all beamline configurations. These were reduced to ±0.2 %/cm by applying optimized BCM profiles. The algorithm converged quickly, validating its underlying assumption that the optimal SOBP profile can be determined by considering only a single dose values at the depths of maximum individual Bragg peak dose. Optimized BCM profiles adequately reduce SOBP slope in the treatment field, and may be further studied using time-resolved MCNPX simulations. The algorithm presented efficiently calculates BCM profiles, fully accounting for beamline scatter and partial-shining effects across adjacent steps on the range modulator wheels, and without requiring direct measurements. Future work includes applying this same algorithm to optimize the SOBP distal dose profile. Partially supported by MEVION Medical Systems, Inc.


Impaired responsiveness of homosexual men with HIV antibodies to plasma derived hepatitis B vaccine.
Thirty five homosexual men (17 positive for antibody to the human immunodeficiency virus (HIV) and 18 consistently negative) were vaccinated against hepatitis B virus infection. Eight of the 17 seropositive patients failed to develop detectable hepatitis B surface antibody within three months of the third injection compared with only one of the 18 seronegative patients (p less than 0.01). HIV infection is prevalent in the developed world in groups at risk for hepatitis B infection and in certain Third World countries where widespread vaccination programmes exist. This study shows the impact that coincident HIV infection may have on an otherwise efficacious vaccine. The efficacy of this and other vaccines in patients infected with HIV needs to be studied urgently.


Design principle of Au grating couplers for quantum-well infrared photodetectors.
Based on analytic formulas and numerical simulations, we investigate the enhancement effect of Au gratings with spoof plasmon resonances on quantum-well infrared photodetectors (QWIPs) operating between 2 and 30 μm. It is found that a simple analytic formula can well estimate the resonant wavelengths of Au gratings. Using optimal grating parameters, the absorption in the QW active region can be enhanced by 4-5 times compared with that in the reference structure (without gratings and with an isotropic active region). For s-polarized light, a high enhancement (>1.4) can occur in a broad range of incident angle (|θ|<40°).


Investigation of the halophilic PET hydrolase PET6 from Vibrio gazogenes.
The handling of plastic waste and the associated ubiquitous occurrence of microplastic poses one of the biggest challenges of our time. Recent investigations of plastic degrading enzymes have opened new prospects for biological microplastic decomposition as well as recycling applications. For polyethylene terephthalate, in particular, several natural and engineered enzymes are known to have such promising properties. From a previous study that identified new PETase candidates by homology search, we chose the candidate PET6 from the globally distributed, halophilic organism Vibrio gazogenes for further investigation. By mapping the occurrence of Vibrios containing PET6 homologs we demonstrated their ubiquitous prevalence in the pangenome of several Vibrio strains. The biochemical characterization of PET6 showed that PET6 has a comparatively lower activity than other enzymes but also revealed a superior turnover at very high salt concentrations. The crystal structure of PET6 provides structural insights into this adaptation to saline environments. By grafting only a few beneficial mutations from other PET degrading enzymes onto PET6, we increased the activity up to three-fold, demonstrating the evolutionary potential of the enzyme. MD simulations of the variant helped rationalize the mutational effects of those mutants and elucidate the interaction of the enzyme with a PET substrate. With tremendous amounts of plastic waste in the Ocean and the prevalence of Vibrio gazogenes in marine biofilms and estuarine marshes, our findings suggest that Vibrio and the PET6 enzyme are worthy subjects to study the PET degradation in marine environments.


Transfer of innovation on allergic rhinitis and asthma multimorbidity in the elderly (MACVIA-ARIA) - EIP on AHA Twinning Reference Site (GARD research demonstration project).
The overarching goals of the European Innovation Partnership on Active and Healthy Ageing (EIP on AHA) are to enable European citizens to lead healthy, active and independent lives whilst ageing. The EIP on AHA includes 74 Reference Sites. The aim of this study was to transfer innovation from an app developed by the MACVIA-France EIP on AHA reference site (Allergy Diary) to other reference sites. The phenotypic characteristics of rhinitis and asthma multimorbidity in adults and the elderly will be compared using validated information and communication technology (ICT) tools (i.e. the Allergy Diary and CARAT: Control of Allergic Rhinitis and Asthma Test) in 22 Reference Sites or regions across Europe. This will improve the understanding, assessment of burden, diagnosis and management of rhinitis in the elderly by comparison with an adult population. Specific objectives will be: (i) to assess the percentage of adults and elderly who are able to use the Allergy Diary, (ii) to study the phenotypic characteristics and treatment over a 1-year period of rhinitis and asthma multimorbidity at baseline (cross-sectional study) and (iii) to follow-up using visual analogue scale (VAS). This part of the study may provide some insight into the differences between the elderly and adults in terms of response to treatment and practice. Finally (iv) work productivity will be examined in adults.


My aortic root simulator: if I can build it, you can build it.
Simulation has become an integral part of thoracic surgical training that has been proven to improve residents' skills. The purpose of the current study was to develop a low-fidelity and low-cost simulator for aortic root surgery that could provide training in multiple aortic valve and root procedures. The current aortic root simulator is made of inexpensive, easily available materials and can be built easily to simulate an aortic root. The basic components are rubber glove, plastic cup, silk tape and small silastic tubes. The simulator was proven to be simple and cost-effective. Several procedures have been performed with this simulator such as standard aortic valve replacement, aortic valve repair, valve-sparing aortic root replacement, and modified Bentall procedure. The building of the simulator is in itself a beneficial process to the resident. The benefit of its construction is a great learning experience. The simulator was placed in a box to create a portable device that can be used by the resident anywhere. This is a simple and cost-effective aortic root simulator that can be built by the thoracic surgical resident to provide training in multiple aortic valve and aortic root procedures. Such low-fidelity portable simulators are beneficial to young trainees and may contribute to improvement of technical skills and procedural knowledge that ultimately leads to improved performance in the operative field.


Association of Vitamin D Status with the Severity and Mortality of Community-Acquired Pneumonia in Iran during 2016-2017: A Prospective Cohort Study.
Community-acquired pneumonia (CAP) is a common disease considered as a major public health problem. It causes considerable morbidity and mortality despite antibiotic treatments. Hospital admission of CAP patients is a significant financial burden and many efforts are ongoing to decrease hospital stay durations. Vitamin D deficiency is associated with increased risk of respiratory infections. This study was designed to determine the association of vitamin D status with hospitalized CAP patient mortality and disease severity. This prospective cohort study examined 180 CAP patients admitted to a teaching Hospital in Tehran, Iran during 2016-2017. Their demographic and anthropometric characteristics were recorded. The disease severity was evaluated based on CURB-65. Vitamin D status was determined by measuring by serum 25-hydroxylated vitamin D (25(OH)D) with ELISA. The patients were followed for 30 days to evaluate their vitality. One hundred and eighty pneumonia patients, including 104 males and 84 females, were recruited from respiratory disease, infectious disease, emergency, and ICU wards. Nearly 18% of the patients were current smokers. The CAP severity, evaluated by CURB-65, was determined to be non-severe in 74.4% of the patients. Patients were classified as vitamin D sufficient, insufficient, or deficient. Thirty percent of the patients were vitamin D sufficient, 18% were insufficient, and 52% were deficient. Thirty-day mortality was 40% (72 cases). Mortality was greater in males than in females (47.1% vs. 30.3%, p=0.03). The disease was significantly less severe in the patients who survived than in those who did not. The vitamin D status differed between males and females (p=0.027). The vitamin D status was lower in the more severe cases than in the less (p=0.036), and vitamin D deficiency was more prevalent in patients who died than in those who lived. Vitamin D concentration was negatively correlated with hospital stay duration. The 25(OH)D concentration was significantly greater in patients who survived than in those who did not (p<0.001). Pneumonia severity and mortality risk were greater and hospital stays longer in vitamin D-deficient patients than in those with higher vitamin D status.


A proposed core genome scheme for analyses of the Salmonella genus.
The salmonellae are found in a wide range of animal hosts and many food products for human consumption. Most cases of human disease are caused by S. enterica subspecies I; however as opportunistic pathogens the other subspecies (II-VI) and S. bongori are capable of causing disease. Loci that were not consistently present in all of the species and subspecies were removed from a previously proposed core genome scheme (EBcgMLSTv2.0), the removal of these 252 loci resulted in a core genus scheme (SalmcgMLSTv1.0). SalmcgMLSTv1.0 clustered isolates from the same subspecies more rapidly and more accurately grouped isolates from different subspecies when compared with EBcgMLSTv2.0. All loci within the EBcgMLSTv2.0 scheme were present in over 98% of S. enterica subspecies I isolates and should, therefore, continue to be used for subspecies I analyses, while the SalmcgMLSTv1.0 scheme is more appropriate for cross genus investigations.


Coordinating Systems of Care Using Health Information Technology: Development of the ADHD Care Assistant.
Perhaps the two principal venues for the delivery of mental health services are schools and primary care practices. Unfortunately, these systems of care are poorly connected, which may result in care that is fragmented and suboptimal. This article describes the development and implementation of an electronic health record portal, known as the ADHD Care Assistant, to facilitate the sharing of information between schools and primary care offices to promote the use of evidence-based practices for managing children with behavioral health conditions. A feasibility study was conducted across 19 diverse primary care practices with 105 participating providers. Across the practices, 67% of providers activated the system for at least one patient and 32% activated it for five or more cases. Care Assistant use was lower in practices with a relatively high percentage of Medicaid patients (≥ 25%). The article discusses challenges that have arisen among primary care providers, parents, and schools in using the Care Assistant, potential strategies for addressing the challenges, and directions for future research and practice.


A Bayesian network meta-analysis on comparisons of enamel matrix derivatives, guided tissue regeneration and their combination therapies.
Guided tissue regeneration (GTR) and enamel matrix derivatives (EMD) are two popular regenerative treatments for periodontal infrabony lesions. Both have been used in conjunction with other regenerative materials. We conducted a Bayesian network meta-analysis of randomized controlled trials on treatment effects of GTR, EMD and their combination therapies. A systematic literature search was conducted using the Medline, EMBASE, LILACS and CENTRAL databases up to and including June 2011. Treatment outcomes were changes in probing pocket depth (PPD), clinical attachment level (CAL) and infrabony defect depth. Different types of bone grafts were treated as one group and so were barrier membranes. A total of 53 studies were included in this review, and we found small differences between regenerative therapies which were non-significant statistically and clinically. GTR and GTR-related combination therapies achieved greater PPD reduction than EMD and EMD-related combination therapies. Combination therapies achieved slightly greater CAL gain than the use of EMD or GTR alone. GTR with BG achieved greatest defect fill. Combination therapies performed better than single therapies, but the additional benefits were small. Bayesian network meta-analysis is a promising technique to compare multiple treatments. Further analysis of methodological characteristics will be required prior to clinical recommendations.


A Bayesian mixture model for across-site heterogeneities in the amino-acid replacement process.
Most current models of sequence evolution assume that all sites of a protein evolve under the same substitution process, characterized by a 20 x 20 substitution matrix. Here, we propose to relax this assumption by developing a Bayesian mixture model that allows the amino-acid replacement pattern at different sites of a protein alignment to be described by distinct substitution processes. Our model, named CAT, assumes the existence of distinct processes (or classes) differing by their equilibrium frequencies over the 20 residues. Through the use of a Dirichlet process prior, the total number of classes and their respective amino-acid profiles, as well as the affiliations of each site to a given class, are all free variables of the model. In this way, the CAT model is able to adapt to the complexity actually present in the data, and it yields an estimate of the substitutional heterogeneity through the posterior mean number of classes. We show that a significant level of heterogeneity is present in the substitution patterns of proteins, and that the standard one-matrix model fails to account for this heterogeneity. By evaluating the Bayes factor, we demonstrate that the standard model is outperformed by CAT on all of the data sets which we analyzed. Altogether, these results suggest that the complexity of the pattern of substitution of real sequences is better captured by the CAT model, offering the possibility of studying its impact on phylogenetic reconstruction and its connections with structure-function determinants.


New insights into the WalK/WalR (YycG/YycF) essential signal transduction pathway reveal a major role in controlling cell wall metabolism and biofilm formation in Staphylococcus aureus.
The highly conserved WalK/WalR (also known as YycG/YycF) two-component system is specific to low-G+C gram-positive bacteria. While this system is essential for cell viability, both the nature of its regulon and its physiological role have remained mostly uncharacterized. We observed that, unexpectedly, Staphylococcus aureus cell death induced by WalKR depletion was not followed by lysis. We show that WalKR positively controls autolytic activity, in particular that of the two major S. aureus autolysins, AtlA and LytM. By using our previously characterized consensus WalR binding site and carefully reexamining the genome annotations, we identified nine genes potentially belonging to the WalKR regulon that appeared to be involved in S. aureus cell wall degradation. Expression of all of these genes was positively controlled by WalKR levels in the cell, leading to high resistance to Triton X-100-induced lysis when the cells were starved for WalKR. Cells lacking WalKR were also more resistant to lysostaphin-induced lysis, suggesting modifications in cell wall structure. Indeed, lowered levels of WalKR led to a significant decrease in peptidoglycan biosynthesis and turnover and to cell wall modifications, which included increased peptidoglycan cross-linking and glycan chain length. We also demonstrated a direct relationship between WalKR levels and the ability to form biofilms. This is the first example in S. aureus of a regulatory system positively controlling autolysin synthesis and biofilm formation. Taken together, our results now define this signal transduction pathway as a master regulatory system for cell wall metabolism, which we have accordingly renamed WalK/WalR to reflect its true function.


Patient-centered digital biomarkers for allergic respiratory diseases and asthma: The ARIA-EAACI approach - ARIA-EAACI Task Force Report.
Biomarkers for the diagnosis, treatment and follow-up of patients with rhinitis and/or asthma are urgently needed. Although some biologic biomarkers exist in specialist care for asthma, they cannot be largely used in primary care. There are no validated biomarkers in rhinitis or allergen immunotherapy (AIT) that can be used in clinical practice. The digital transformation of health and health care (including mHealth) places the patient at the center of the health system and is likely to optimize the practice of allergy. Allergic Rhinitis and its Impact on Asthma (ARIA) and EAACI (European Academy of Allergy and Clinical Immunology) developed a Task Force aimed at proposing patient-reported outcome measures (PROMs) as digital biomarkers that can be easily used for different purposes in rhinitis and asthma. It first defined control digital biomarkers that should make a bridge between clinical practice, randomized controlled trials, observational real-life studies and allergen challenges. Using the MASK-air app as a model, a daily electronic combined symptom-medication score for allergic diseases (CSMS) or for asthma (e-DASTHMA), combined with a monthly control questionnaire, was embedded in a strategy similar to the diabetes approach for disease control. To mimic real-life, it secondly proposed quality-of-life digital biomarkers including daily EQ-5D visual analogue scales and the bi-weekly RhinAsthma Patient Perspective (RAAP). The potential implications for the management of allergic respiratory diseases were proposed.


Oral contraceptives and breast cancer risk among younger women.
Several studies have suggested a link between oral contraceptive use and breast cancer in younger women, but it is possible that chance or bias, including selective screening of contraceptive users, contributed to the putative association. Given that oral contraceptives were first marketed in the United States in the early 1960s, we conducted a population-based case-control study to examine the relationship between use of oral contraceptives and breast cancer among women in a recently assembled cohort, focusing on women younger than 45 years of age who had the opportunity for exposure throughout their entire reproductive years. Breast cancer patients and healthy control subjects were identified, the latter group by random-digit dialing, in Atlanta, Ga., Seattle/Puget Sound, Wash., and central New Jersey. In Seattle and New Jersey, the study was confined to women 20 through 44 years of age; in Atlanta the age range was extended through 54 years. Patients included women with in situ or invasive breast cancer newly diagnosed during the period of May 1, 1990, through December 31, 1992. In-person interviews were completed by 2203 (86.4%) of 2551 eligible patients and 2009 (78.1%) of 2571 eligible control subjects. Analyses focused on women younger than 45 years of age (1648 patients and 1505 control subjects) to maximize opportunities for extended exposure. Logistic regression analyses were used to obtain maximum likelihood estimates of relative risks (RRs) and their 95% confidence intervals (CIs). Among women younger than 45 years, oral contraceptive use for 6 months or longer was associated with an RR for breast cancer of 1.3 (95% CI = 1.1-1.5). Risks were enhanced for breast cancers occurring prior to age 35 years (RR = 1.7; 95% CI = 1.2-2.6), with the RR rising to 2.2 (95% CI = 1.2-4.1) for users of 10 or more years. The RR for breast cancer for those whose oral contraceptive use began early (before age 18 years) and continued long-term (> 10 years) was even higher (RR = 3.1; 95% CI = 1.4-6.7). The RRs observed for those who used oral contraceptives within 5 years of cancer diagnosis were higher than for those who had not, with the effect most marked for women younger than age 35 years (RR = 2.0; 95% CI = 1.3-3.1). Oral contraceptive associations were also strongest for cancers diagnosed at advanced stages. Evaluation of screening histories and methods of diagnosis failed to support the speculation that associations could be due to selective screening. Among women 45 years of age and older, no associations of risk with use of oral contraceptives were noted. The relationship between oral contraceptives and breast cancer in young women appears to have a biologic basis rather than to be an artifact or the result of bias.


Ideal femoral head size in total hip arthroplasty balances stability and volumetric wear.
Over the last several years, a trend towards increasing femoral head size in total hip arthroplasty to improve stability and impingement free range of motion has been observed. The specific questions we sought to answer in our review were: (1) What are the potential advantages and disadvantages of metal-on-metal, ceramic-on-ceramic, and metal-on-polyethylene bearings? (2) What is effect that femoral head size has on joint kinematics? (3) What is the effect that large femoral heads have on bearing surface wear? A PubMed search and a review of 2012 Orthopaedic Research Society abstracts was performed and articles were chosen that directly answered components of the specific aims and that reported outcomes with contemporary implant designs or materials. A review of the literature suggests that increasing femoral head size decreases the risk of postoperative dislocation and improves impingement free range of motion; however, volumetric wear increases with large femoral heads on polyethylene and increases corrosion of the stem in large metal-on-metal modular total hip arthroplasty (THA); however, the risk of potentially developing osteolysis or adverse reactions to metal debris respectively is still unknown. Further, the effect of large femoral heads with ceramic-on-ceramic THA is unclear, due to limited availability and published data. Surgeons must balance the benefits of larger head size with the increased risk of volumetric wear when determining the appropriate head size for a given patient.


Deformation of Microporous Carbons during N2, Ar, and CO2 Adsorption: Insight from the Density Functional Theory.
Using the nonlocal density functional theory, we investigate adsorption of N2 (77 K), Ar (77 K), and CO2 (273 K) and respective adsorption-induced deformation of microporous carbons. We show that the smallest micropores comparable in size and even smaller than the nominal molecular diameter of the adsorbate contribute significantly to the development of the adsorption stress. While pores of approximately the nominal adsorbate diameter exhibit no adsorption stress regardless of their filling level, the smaller pores cause expansive adsorption stresses up to almost 4 GPa. Accounting for this effect, we determined the pore-size distribution of a synthetic microporous carbon by simultaneously fitting its experimental CO2 adsorption isotherm (273 K) and corresponding adsorption-induced strain measured by in situ dilatometry. Based on the pore-size distribution and the elastic modulus fitted from CO2 data, we predicted the sample's strain isotherms during N2 and Ar adsorption (77 K), which were found to be in reasonable agreement with respective experimental data. The comparison of calculations and experimental results suggests that adsorption-induced deformation caused by micropores is not limited to the low relative pressures typically associated with the micropore filling, but is effective over the whole relative pressure range up to saturation pressure.


Better Utilization of Mouse Models of Neurodegenerative Diseases in Preclinical Studies: From the Bench to the Clinic.
The major symptom of Alzheimer's disease is dementia progressing with age. Its clinical diagnosis is preceded by a long prodromal period of brain pathology that encompasses both formation of extracellular amyloid and intraneuronal tau deposits in the brain and widespread neuronal death. At present, familial cases of dementia provide the most promising foundation for modeling neurodegenerative tauopathies, a group of heterogeneous disorders characterized by prominent intracellular accumulation of hyperphosphorylated tau protein. In this chapter, we describe major behavioral hallmarks of tauopathies, briefly outline the genetics underlying familial cases, and discuss the arising implications for modeling the disease in transgenic mouse systems. The selection of tests performed to evaluate the phenotype of a model should be guided by the key behavioral hallmarks that characterize human disorder and their homology to mouse cognitive systems. We attempt to provide general guidelines and establish criteria for modeling dementia in a mouse; however, interpretations of obtained results should avoid a reductionist "one gene, one disease" explanation of model characteristics. Rather, the focus should be directed to the question of how the mouse genome can cope with the over-expression of the protein coded by transgene(s). While each model is valuable within its own constraints and the experiments performed are guided by specific hypotheses, we seek to expand upon their methodology by offering guidance spanning from issues of mouse husbandry to choices of behavioral tests and routes of drug administration that might increase the external validity of studies and consequently optimize the translational aspect of preclinical research.


A randomised study in healthy volunteers to investigate the safety, tolerability and pharmacokinetics of idarucizumab, a specific antidote to dabigatran.
Idarucizumab, a monoclonal antibody fragment that binds dabigatran with high affinity, is in development as a specific antidote for dabigatran. In this first-in-human, single-rising-dose study, we investigated the pharmacokinetics, safety and tolerability of idarucizumab. Healthy male volunteers aged 18-45 years received between 20 mg and 8 g idarucizumab as a 1-hour intravenous infusion in 10 sequential dose groups, or 1, 2 or 4 g idarucizumab as a 5-minute infusion. Subjects within each dose group were randomised 3:1 to idarucizumab or placebo. A total of 110 randomised subjects received study drug (27 placebo, 83 idarucizumab). Peak and total exposure to idarucizumab increased proportionally with dose. Maximum plasma concentrations were achieved near the end of infusion, followed by a rapid decline, with an initial idarucizumab half-life of ~45 minutes. For the 5-minute infusions, this resulted in a reduction of plasma concentrations to less than 5 % of peak within 4 hours. Idarucizumab (in the absence of dabigatran) had no effect on coagulation parameters or endogenous thrombin potential. Overall adverse event (AE) frequency was similar for idarucizumab and placebo, and no relationship with idarucizumab dose was observed. Drug-related AEs (primary endpoint) were rare (occurring in 2 placebo and 3 idarucizumab subjects) and were mostly of mild intensity; none of them resulted in study discontinuation. In conclusion, the pharmacokinetic profile of idarucizumab meets the requirement for rapid peak exposure and rapid elimination, with no effect on pharmacodynamic parameters. Idarucizumab was safe and well tolerated in healthy males.


Indoor Air Quality and Sick Building Syndrome: Are Green Buildings Better than Conventional Buildings?
Indoor air quality (IAQ) influences human health, productivity and wellness. Green buildings are believed to have better IAQ. The 'sick building syndrome' (SBS) describes a set of nonspecific symptoms experienced by occupants due to time spent in a building with poor IAQ. Thus this study was undertaken to assess the IAQ in green buildings and compare it with that of conventional buildings. The prevalence of SBS in both types of buildings is also studied. In five pairs of green and conventional buildings measurements of comfort parameters (temperature & relative humidity) and indoor air pollutants using monitors was done. 148 employees which included 84 from green buildings and 64 from conventional buildings were surveyed for SBS using an interviewer-administered questionnaire. The analysis was done using SPSS16 and included Mann Whitney for IAQ pollutant concentrations and Chi-square for the SBS prevalence. Similar indoor air quality was found in both types of buildings. The mean of temperature, CO2 and formaldehyde was statistically lower in green buildings. The SBS prevalence was found to be 38.1% in green buildings and 53.1% in conventional buildings. Thus to conclude the poorly maintained green building does not have any added advantage for occurrence of SBS.


Diagnosis and treatment planning for the surgical-orthodontic patient.
Contemporary orthognathic surgery has made it possible to reposition either or both jaws in one piece or as segmental osteotomies. This has provided the patient, who has a severe dentofacial problem, with the option of having all components of their malocclusion and facial aesthetic concerns addressed. The close collaboration between the orthodontist and the surgeon provides a comprehensive diagnosis and treatment plan to be generated. The goal of the presurgical phase of orthodontics is to remove the dental compensations and allow optimum surgical correction of the jaw discrepancy. A short phase of orthodontic treatment postoperatively is usually necessary to detail the final occlusion. A systematic approach to diagnosis and treatment planning is discussed in the context of the problem-oriented approach. The application of this approach is illustrated in a case report of the combined orthodontic-surgical treatment of a patient who benefited from a two-jaw surgical procedure with pre- and postoperative orthodontic treatment. The principles of presurgical orthodontic treatment are discussed and should avoid any counterproductive tooth movements that can be part of the surgical correction. Tooth movements done as part of the postoperative treatment should avoid compromising the surgical outcome. Illustrations of the cephalometric prediction tracing and its application to the diagnosis and treatment plan of the patient discussed in the case report is an important aspect in providing the dentofacial patient with a representation of the expected treatment outcome.


Autophagy and aging: Maintaining the proteome through exercise and caloric restriction.
Accumulation of dysfunctional and damaged cellular proteins and organelles occurs during aging, resulting in a disruption of cellular homeostasis and progressive degeneration and increases the risk of cell death. Moderating the accrual of these defunct components is likely a key in the promotion of longevity. While exercise is known to promote healthy aging and mitigate age-related pathologies, the molecular underpinnings of this phenomenon remain largely unclear. However, recent evidences suggest that exercise modulates the proteome. Similarly, caloric restriction (CR), a known promoter of lifespan, is understood to augment intracellular protein quality. Autophagy is an evolutionary conserved recycling pathway responsible for the degradation, then turnover of cellular proteins and organelles. This housekeeping system has been reliably linked to the aging process. Moreover, autophagic activity declines during aging. The target of rapamycin complex 1 (TORC1), a central kinase involved in protein translation, is a negative regulator of autophagy, and inhibition of TORC1 enhances lifespan. Inhibition of TORC1 may reduce the production of cellular proteins which may otherwise contribute to the deleterious accumulation observed in aging. TORC1 may also exert its effects in an autophagy-dependent manner. Exercise and CR result in a concomitant downregulation of TORC1 activity and upregulation of autophagy in a number of tissues. Moreover, exercise-induced TORC1 and autophagy signaling share common pathways with that of CR. Therefore, the longevity effects of exercise and CR may stem from the maintenance of the proteome by balancing the synthesis and recycling of intracellular proteins and thus may represent practical means to promote longevity.


Full solution processed mesostructured optical resonators integrating colloidal semiconductor quantum dots.
Herein we show a solution based synthetic pathway to obtain a resonant optical cavity with embedded colloidal semiconductor quantum dots (CSQDs). The optical cavity pore network, surrounded by two dense Bragg mirrors, was designed ad hoc to selectively host the quantum dots, while uncontrolled infiltration of those in the rest of the layered structure was prevented. Coupling between the optical resonant modes of the host and the natural emission of the embedded nanoparticles gives rise to the fine tuning of the luminescence spectrum extracted from the ensemble. Our approach overcomes, without the need for an encapsulating agent and exclusively by solution processing, the difficulties that arise from the low thermal and chemical stability of the CSQDs. It opens the route to achieving precise control over their location and hence over the spectral properties of light emitted by these widely employed nanomaterials. Furthermore, as the porosity of the cavity is preserved after infiltration, the system remains responsive to environmental changes, which provides an added value to the proposed structure.


Cooperating in the face of uncertainty: a consistent framework for understanding the evolution of cooperation.
The evolution of cooperative behaviour, whereby individuals enhance the fitness of others at an apparent cost to themselves, represents one of the greatest paradoxes of evolution. Individuals that engage in such cooperative behaviour can, however, be favoured by natural selection if cooperative actions confer higher fitness than alternative actions. To understand the evolution of cooperative behaviour, the direct and indirect genetic benefits that individuals accrue in the present and future must be summed - this can be accomplished without any reference to the colorful vocabulary typically associated with studies of cooperation. When benefits are accrued indirectly through relatives or directly in the future individuals must be able to assess and enhance their probability of accruing those benefits and behave accordingly. We suggest that, in the same way that studies of kin recognition systems improved our understanding of how individuals assess and enhance their probability of accruing indirect benefits, studies of various forms of inheritance and reciprocation recognition systems will improve our understanding of how individuals assess and enhance their probability of accruing future benefits. Recognizing the parallel between studies of indirect fitness and future fitness, at multiple levels of analysis, will move us toward a simpler and more consistent framework for understanding the evolution of cooperative behaviour.


Pilot study of pegylated interferon alfa-2b and ribavirin for recurrent hepatitis C after liver transplantation.
Recurrent hepatitis C is often treated with an interferon and ribavirin combination therapy, but the results have been disappointing. Given the promising results reported with pegylated interferon and ribavirin for hepatitis C, we were interested in evaluating the effectiveness of this treatment in liver transplant recipients with recurrent hepatitis C (HCV). Between November 2001 and September 2002, patients with recurrent HCV were screened to determine if they were eligible for treatment. Liver function tests, HCV-RNA, and liver biopsies were performed on all patients prior to treatment. HCV-RNA was repeated at 3 months, the end of treatment (EOT), and 6 months after EOT for patients who were HCV-RNA negative at EOT. Patients were prospectively followed after starting weekly pegylated interferon alfa-2b 1.5 mcg/kg per week and ribavirin 800 mg per day (Schering-Plough, Kenilworth, NJ, USA) with folic acid 1 mg per day. Thirty-nine patients eligible for treatment displayed a median age of 50.4 years. Eighteen patients completed treatment, 4 remain on treatment, and 17 were intolerant. Sustained HCV-RNA eradication occurred in 66.7% of patients who completed treatment. Side effects led to treatment withdrawal in 17 patients (43.6%) In an intention-to treat analysis, sustained HCV-RNA eradication occurred in 30.8% of patients. Side effects are an important limiting factor in the treatment of recurrent HCV with pegylated interferon and ribavirin. However, these results are encouraging as sustained HCV eradication occurred in at least 66.7% of patients who completed treatment. Prospective randomized trials are required to assess the effectiveness of this treatment and its impact on quality of life and histology.


LKB1, a novel serine/threonine protein kinase and potential tumour suppressor, is phosphorylated by cAMP-dependent protein kinase (PKA) and prenylated in vivo.
Peutz-Jeghers syndrome (PJS) is an autosomal dominant disease characterized by melanocytic macules, hamartomatous polyps and an increased risk for numerous cancers. The human LKB1 (hLKB1) gene encodes a serine/threonine protein kinase that is deficient in the majority of patients with PJS. The murine LKB1 (mLKB1) cDNA was isolated, sequenced and shown to produce a 2.4-kb transcript encoding a 436 amino acid protein with 90% identity with hLKB1. RNA blot and RNase-protection analysis revealed that mLKB1 mRNA is expressed in all tissues and cell lines examined. The widespread expression of LKB1 transcripts is consistent with the elevated risk of multiple cancer types in PJS patients. The predicted LKB1 protein sequence terminates with a conserved prenylation motif (Cys(433)-Lys-Gln-Gln(436)) directly downstream from a consensus cAMP-dependent protein kinase (PKA) phosphorylation site (Arg(428)-Arg-Leu-Ser(431)). The expression of enhanced green fluorescent protein (EGFP)-mLKB1 chimaeras demonstrated that LKB1 possesses a functional prenylation motif that is capable of targeting EGFP to cellular membranes. Mutation of Cys(433) to an alanine residue, but not phosphorylation by PKA, blocked membrane localization. These findings suggest that PKA does phosphorylate LKB1, although this phosphorylation does not alter the cellular localization of LKB1.


Recurrent Optic Neuritis Associated With MOG Antibody Seropositivity.
Optic neuritis (ON) can be the first manifestation of autoimmune central nervous system diseases such as multiple sclerosis and neuromyelitis optica, but it can also occur as an isolated monophasic or relapsing disease. A proportion of these patients have antibodies against myelin oligodendrocyte glycoprotein (MOG). We report a case of recurrent ON with MOG antibodies in a South Asian patient. A 50-year-old Sri Lankan female with a previous history of 2 episodes of steroid-responsive ON presented with a 20/120 visual impairment of her right eye. On examination on admission, visual acuity was noted to be 20/30 on the left and visual evoked potentials were prolonged bilaterally, but all other neurological investigations including screening tests for infections and vasculitis were unremarkable. AQP4 antibodies were negative but MOG antibodies were positive. She responded to intravenous and long-term steroids combined with mycophenolate mofetil, but relapsed when steroids were discontinued. She has been relapse free on low-dose steroids and mycophenolate for 2 years. Recurrent ON occurs in association with MOG antibodies distinct from neuromyelitis optica and multiple sclerosis, and may require steroid-based immunosuppression to maintain remission. This case report from South Asia adds to map the worldwide occurrence of MOG antibody disease.


Blood arsenic as a biomarker of arsenic exposure: results from a prospective study.
Exposure to arsenic (As)-contaminated drinking water affects millions of people worldwide. Arsenic exposure is associated with skin lesions, skin, lung, kidney and liver cancers, neurologic and cardiovascular effects. Past studies involving biomarkers of As exposure have typically examined urinary As (UAs) (adjusted for urinary creatinine), hair or toenail As, but not blood As (BAs) since blood concentrations are exceedingly low and are not detectable by conventional atomic absorption spectrophotometric techniques. In a case-cohort analysis of 303 newly diagnosed cases of skin lesions, and 849 subcohort members randomly selected from 8092 participants in the health effects of as longitudinal study (HEALS) in Araihazar, Bangladesh, we measured blood, urine and water As concentrations, and examined their associations with each other, and with the risk for skin lesions. BAs concentrations were highly correlated with creatinine-adjusted UAs concentrations (r=0.85) and with water As (WAs) (r=0.75). We observed consistent dose-response relationships between the risk of skin lesions and all the measures of As exposure. Rate ratios (RRs) for skin lesions by quintile of As exposure, adjusted for age and gender, revealed that the two highest quintiles were significantly related to an increased risk of skin lesions for each measure of exposure: BAs, UAs, WAs and a time-weighted water As variable. This prospective study confirms the increased risk of skin lesions in relation to As concentrations in blood, urine and water and also establishes that BAs is a useful biomarker of As exposure in this study population.


Commitment to the regulatory T cell lineage requires CARMA1 in the thymus but not in the periphery.
Regulatory T (T(reg)) cells expressing forkhead box P3 (Foxp3) arise during thymic selection among thymocytes with modestly self-reactive T cell receptors. In vitro studies suggest Foxp3 can also be induced among peripheral CD4(+) T cells in a cytokine dependent manner. T(reg) cells of thymic or peripheral origin may serve different functions in vivo, but both populations are phenotypically indistinguishable in wild-type mice. Here we show that mice with a Carma1 point mutation lack thymic CD4(+)Foxp3(+) T(reg) cells and demonstrate a cell-intrinsic requirement for CARMA1 in thymic Foxp3 induction. However, peripheral Carma1-deficient T(reg) cells could be generated and expanded in vitro in response to the cytokines transforming growth factor beta (TGFbeta) and interleukin-2 (IL-2). In vivo, a small peripheral T(reg) pool existed that was enriched at mucosal sites and could expand systemically after infection with mouse cytomegalovirus (MCMV). Our data provide genetic evidence for two distinct mechanisms controlling regulatory T cell lineage commitment. Furthermore, we show that peripheral T(reg) cells are a dynamic population that may expand to limit immunopathology or promote chronic infection.


Drug delivery through the skin: molecular simulations of barrier lipids to design more effective noninvasive dermal and transdermal delivery systems for small molecules, biologics, and cosmetics.
The delivery of drugs through the skin provides a convenient route of administration that is often preferable to injection because it is noninvasive and can typically be self-administered. These two factors alone result in a significant reduction of medical complications and improvement in patient compliance. Unfortunately, a significant obstacle to dermal and transdermal drug delivery alike is the resilient barrier that the epidermal layers of the skin, primarily the stratum corneum, presents for the diffusion of exogenous chemical agents. Further advancement of transdermal drug delivery requires the development of novel delivery systems that are suitable for modern, macromolecular protein and nucleotide therapeutic agents. Significant effort has already been devoted to obtain a functional understanding of the physical barrier properties imparted by the epidermis, specifically the membrane structures of the stratum corneum. However, structural observations of membrane systems are often hindered by low resolutions, making it difficult to resolve the molecular mechanisms related to interactions between lipids found within the stratum corneum. Several models describing the molecular diffusion of drug molecules through the stratum corneum have now been postulated, where chemical permeation enhancers are thought to disrupt the underlying lipid structure, resulting in enhanced permeability. Recent investigations using biphasic vesicles also suggested a possibility for novel mechanisms involving the formation of complex polymorphic lipid phases. In this review, we discuss the advantages and limitations of permeation-enhancing strategies and how computational simulations, at the atomic scale, coupled with physical observations can provide insight into the mechanisms of diffusion through the stratum corneum.


A novel X-linked disorder of immune deficiency and hypohidrotic ectodermal dysplasia is allelic to incontinentia pigmenti and due to mutations in IKK-gamma (NEMO).
Hypohidrotic ectodermal dysplasia (HED), a congenital disorder of teeth, hair, and eccrine sweat glands, is usually inherited as an X-linked recessive trait, although rarer autosomal dominant and recessive forms exist. We have studied males from four families with HED and immunodeficiency (HED-ID), in which the disorder segregates as an X-linked recessive trait. Affected males manifest dysgammaglobulinemia and, despite therapy, have significant morbidity and mortality from recurrent infections. Recently, mutations in IKK-gamma (NEMO) have been shown to cause familial incontinentia pigmenti (IP). Unlike HED-ID, IP affects females and, with few exceptions, causes male prenatal lethality. IKK-gamma is required for the activation of the transcription factor known as "nuclear factor kappa B" and plays an important role in T and B cell function. We hypothesize that "milder" mutations at this locus may cause HED-ID. In all four families, sequence analysis reveals exon 10 mutations affecting the carboxy-terminal end of the IKK-gamma protein, a domain believed to connect the IKK signalsome complex to upstream activators. The findings define a new X-linked recessive immunodeficiency syndrome, distinct from other types of HED and immunodeficiency syndromes. The data provide further evidence that the development of ectodermal appendages is mediated through a tumor necrosis factor/tumor necrosis factor receptor-like signaling pathway, with the IKK signalsome complex playing a significant role.


Inhibition of contact-mediated activation of factor XI protects baboons against S aureus-induced organ damage and death.
Staphylococcus aureus infections can produce systemic bacteremia and inflammation in humans, which may progress to severe sepsis or septic shock, even with appropriate antibiotic treatment. Sepsis may be associated with disseminated intravascular coagulation and consumptive coagulopathy. In some types of mouse infection models, the plasma coagulation protein factor XI (FXI) contributes to the pathogenesis of sepsis. We hypothesize that FXI also contributes to the pathogenesis of sepsis in primates, and that pharmacological interference with FXI will alter the outcome of Staphylococcus aureus-induced lethality in a baboon model. Pretreatment of baboons with the anti-FXI antibody 3G3, a humanized variant of the murine monoclonal 14E11 that blocks FXI activation by FXIIa, substantially reduced the activation of coagulation, as reflected by clotting times and plasma complexes of coagulation proteases (FXIIa, FXIa, FIXa, FXa, FVIIa, and thrombin) with serpins (antithrombin or C1 inhibitor) following infusion of heat-inactivated S aureus 3G3 treatment reduced fibrinogen and platelet consumption, fibrin deposition in tissues, neutrophil activation and accumulation in tissues, cytokine production, kininogen cleavage, cell death, and complement activation. Overall, 3G3 infusion protected the structure and function of multiple vital organs, including lung, heart, liver, and kidney. All treated animals reached the end point survival (7 days), whereas all nontreated animals developed terminal organ failure within 28 hours. We conclude that FXI plays a role in the pathogenesis of S aureus-induced disseminated intravascular coagulation and lethality in baboons. The results provide proof of concept for future therapeutic interventions that may prevent sepsis-induced organ failure and save lives in certain forms of sepsis.


Differential regulation of iron regulatory element-binding protein(s) in cell extracts of activated lymphocytes versus monocytes-macrophages.
The intracellular iron level exerts a negative feedback on transferrin receptor (TfR) expression in cells requiring iron for their proliferation, in contrast to the positive feedback observed in monocytes-macrophages. It has been suggested recently that modulation of TfR and ferritin synthesis by iron is mediated through a cytoplasmic protein(s) (iron regulatory element-binding protein(s) (IRE-BP)), which interacts with ferritin and TfR mRNA at the level of hairpin structures (IRE), thus leading to inhibition of transferrin mRNA degradation and repression of ferritin mRNA translation. In the present study we have evaluated in parallel the level of TfR expression, ferritin, and IRE-BP in cultures of: (i) circulating human lymphocytes stimulated to proliferate by phytohemagglutinin (PHA) and (ii) circulating human monocytes maturing in vitro to macrophages. The cells were grown in either standard or iron-supplemented culture. TfR and ferritin expression was evaluated at both the protein and mRNA level. IRE-BP activity was measured by gel retardation assay in the absence or presence of beta-mercaptoethanol (spontaneous or total IRE-BP activity, respectively). Spontaneous IRE-BP activity, already present at low level in quiescent T lymphocytes, shows a gradual and marked increase in PHA-stimulated T cells from day 1 of culture onward. This increase is directly and strictly correlated with the initiation and gradual rise of TfR expression, which is in turn associated with a decrease of ferritin content. Both the rise of TfR and spontaneous IRE-BP activity are completely inhibited in iron-supplemented T cell cultures. In contrast, the total IRE-BP level is similar in both quiescent and PHA-stimulated lymphocytes, grown in cultures supplemented or not with iron salts. Monocytes maturing in vitro to macrophages show a sharp increase of spontaneous and, to a lesser extent, total IRE-BP; the addition of iron moderately stimulates the spontaneous IRE-BP activity but not the total one. Here again, the rise of spontaneous IRE-BP from very low to high activity is strictly related to the parallel increase of TfR expression and, suprisingly, also with a very pronounced rise of ferritin expression observed at both the mRNA and protein level. It is noteworthy the effect of beta-mercaptoethanol is cell specific, i.e. the ratio of total versus spontaneous IRE-BP activity is different in activated lymphocytes and maturing monocytes.(ABSTRACT TRUNCATED AT 400 WORDS)


Cerebral Venous Sinus Thrombosis in Users of a Hormonal Vaginal Ring.
Oral contraceptive pills (OCPs) are a well-established precipitant of thrombosis. The local release of hormone from the vaginal ring still results in systemic administration and therefore predisposes users to increased risk of venous thrombosis. Although representing only 1% of strokes, cerebral venous sinus thrombosis is eight times more likely in women taking OCPs than in nonusers, and the incidence of cerebral venous sinus thrombosis associated with the vaginal ring is unknown but is likely to be increased. We report four cases of cerebral venous sinus thrombosis in women using hormonal vaginal rings with negative thrombophilia testing who presented to our facility over a 2-year period. Three of the four women required mechanical thrombectomy in addition to anticoagulation, but all four women recovered and were discharged home with minimal or no neurologic deficits. Increased recognition of the thrombogenic potential of hormonal vaginal rings may promote earlier venous imaging and result in better clinical outcomes.


Possible Interplay Between Hospital and Community Transmission of a Novel Clostridium Difficile Sequence Type 295 Recognized by Next-Generation Sequencing.
OBJECTIVE To use next-generation sequencing (NGS) analysis to enhance epidemiological information to identify and resolve a Clostridium difficile outbreak and to evaluate its effectiveness beyond the capacity of current standard PCR ribotyping. METHODS NGS analysis was performed as part of prospective surveillance of all detected C. difficile isolates at a university hospital. An outbreak of a novel C. difficile sequence type (ST)-295 was identified in a hospital and a community hostel for homeless adults. Phylogenetic analysis was performed of all ST-295 and closest ST-2 isolates. Epidemiological details were obtained from hospital records and the public health review of the community hostel. RESULTS We identified 7 patients with C. difficile ST-295 infections between June 2013 and April 2015. Of these patients, 3 had nosocomial exposure to this infection and 3 had possible hostel exposure. Current Society for Healthcare Epidemiology of America (SHEA)- Infectious Diseases Society of America (IDSA) surveillance definitions (2010) were considered in light of our NGS findings. The initial transmission was not detectable using current criteria, because of 16 weeks between ST-295 exposure and symptoms. We included 3 patients with hostel exposure who met surveillance criteria of hospital-acquired infection due to their hospital admissions. CONCLUSION NGS analysis enhanced epidemiological information and helped identify and resolve an outbreak beyond the capacity of standard PCR ribotyping. In this cluster of cases, NGS was used to identify a hostel as the likely source of community-based C. difficile transmission. Infect Control Hosp Epidemiol 2016;37:680-684.


Association of Body Mass Index and Mortality in Japanese Diabetic Men and Women Based on Self-Reports: The Japan Collaborative Cohort (JACC) Study.
The association between body mass index (BMI) and mortality among Asian diabetic people, especially with respect to the obesity paradox (ie, higher BMI is associated with lower mortality risk), remains unresolved. We followed a cohort of 3851 self-reported Japanese diabetics (2115 men and 1736 women) in the Japan Collaborative Cohort Study from 1988-1990 through 2009. Individuals were aged 40 to 79 years and free from a history of cardiovascular disease, cancer, renal disease, or tuberculosis. BMI was grouped into the following four categories: <20.0, 20.0-22.9, 23.0-24.9, and ≥25.0 kg/m2. During 54 707 person-years of follow-up, 1457 deaths from all causes, 445 from cardiovascular disease, 421 from cancer, 43 from renal disease, and 148 from infectious disease were documented. Mortality from all causes, cardiovascular disease, cancer, and renal disease showed L-shaped associations with BMI. Compared to diabetics with BMI of 20.0-22.9 kg/m2, those with BMIs of 23.0-24.9 kg/m2 and ≥25.0 kg/m2 had lower risks of mortality from infectious disease (i.e., obesity paradox). The multivariable HRs for mortality from infectious disease were 0.50 (95% confidence interval, 0.31-0.81) and 0.51 (95% confidence interval, 0.32-0.82) among participants with BMIs of 23.0-24.9 kg/m2 and ≥25.0 kg/m2, respectively. Similar results were observed after stratification by smoking status and age and exclusion of early deaths. We observed L-shaped associations between BMI and mortality from all causes, cardiovascular disease, cancer, and renal disease, while the association between BMI and mortality from infectious disease manifested the obesity paradox.


Possible involvement of monoamines in the release of adipokinetic hormone in the locust Schistocerca gregaria.
1. The adipokinetic hormone release, which can be induced by anticholinesterases, is reduced by depleting the content of monoamines in the nervous system. 2. The participation of monoamines in the pathway of release of adipokinetic hormone is studied in vivo and in vitro. 3. A possible mechanism for anticholinesterase-induced release of this hormone involving cholinergic and aminergic transmission is postulated.


Association between interpregnancy interval and subsequent stillbirth in 58 low-income and middle-income countries: a retrospective analysis using Demographic and Health Surveys.
About 3 million stillbirths occur each year, 98% of which are in low-income and middle-income countries (LMICs). Interpregnancy interval is a key risk factor of interest, because it is modifiable. We aimed to investigate whether there is a causal relationship between the length of interpregnancy interval and risk of subsequent stillbirth. We used Demographic and Health Surveys (2002-18) from 58 LMICs to study reproductive histories of women and to identify livebirths and stillbirths in the preceding 5 years. Countries were selected on the basis of the availability of interpregnancy interval data and other covariates of interest (age, education, urban or rural residence, and wealth) in surveys done since 2002. Exclusion criteria were being nulliparous, having missing parity data, and not having had at least two births (livebirth or stillbirth) in the 5 years before the survey. We combined two analytic approaches: one that analyses intervals between all births and another that analyses intervals within mothers. We report stratified estimates for the first, second, and third intervals, controlling for all past birth outcomes and intervals in a 5-year period, and other socioeconomic covariates. We also explored effect heterogeneity across key cohort subgroups. Between July, 1997, and April, 2018, we identified 716 478 births from 338 223 women in 123 Demographic and Health Surveys from 58 LMICs, of which 9647 were stillbirths. Intervals of less than 6 months were associated with an increased risk of stillbirth in the between-mother models when considering the first interval (risk difference [RD] 0·0096, 95% CI 0·008-0·011). This association was slightly attenuated when considering only the second interval (RD 0·0054, 95% CI 0·0010 to 0·0099) and substantially attenuated when considering only the third interval (0·0007, -0·037 to 0·039). Within-mother modelling showed a null association with intervals of 24-59 months when considering the first and second (RD 0·007, 95% CI -0·001 to 0·016) and first and third (0·040, -0·422 to 0·501) intervals. Although interpregnancy intervals of less than 12 months were associated with increased risk of stillbirth, these effects were attenuated when considering second and third intervals, suggesting the association in the first interval might not be causal. Future studies should use generalisable cohorts with longitudinal data, and report estimates stratified by birth order. Canadian Institutes of Health Research.


Acupuncture treatment modulates the corticostriatal reward circuitry in major depressive disorder.
Major depressive disorder (MDD) is a common disorder with a high prevalence and significant social and economic impacts. Nevertheless, the treatment of MDD is far from satisfactory. Acupuncture treatment has emerged as a promising method for treating MDD. However, the neural mechanism by which acupuncture reduces depressive symptoms is not fully understood. Studies have shown that the corticostriatal reward circuitry is associated with the pathophysiology of MDD; thus, we investigated the corticostriatal resting-state functional connectivity (rsFC) before and after real and sham acupuncture treatments combined with the antidepressant fluoxetine. Forty-six female major depressive patients were assigned to either verum acupuncture plus fluoxetine (n = 22) or sham acupuncture plus fluoxetine (n = 24) treatment for 8 weeks, and resting state functional magnetic resonance imaging (fMRI) data were collected before the first and after the last treatment sessions. The results showed that compared with sham acupuncture, the verum acupuncture group showed: (1) significantly increased rsFC between inferior ventral striatum and medial prefrontal cortex, ventral rostral putamen and amygdala/parahippocampus, as well as dorsal caudate and middle temporal gyrus; (2) significantly decreased rsFC between right ventral rostral putamen and right dorsolateral prefrontal cortex, and right dorsal caudate and bilateral cerebellar tonsil. The increased rsFC between the inferior ventral striatum and medial prefrontal cortex, ventral rostral putamen and amygdala/parahippocampus were significantly positively associated with decreased clinical scores (Montgomery-Åsberg Depression Rating Scale and Self-Rating Depression Scale scores) at the end of the eight-week treatment. Our findings suggest that acupuncture may achieve treatment effects by modulating the corticostriatal reward/motivation circuitry in MDD patients.


Plant species loss affects life-history traits of aphids and their parasitoids.
The consequences of plant species loss are rarely assessed in a multi-trophic context and especially effects on life-history traits of organisms at higher trophic levels have remained largely unstudied. We used a grassland biodiversity experiment and measured the effects of two components of plant diversity, plant species richness and the presence of nitrogen-fixing legumes, on several life-history traits of naturally colonizing aphids and their primary and secondary parasitoids in the field. We found that, irrespective of aphid species identity, the proportion of winged aphid morphs decreased with increasing plant species richness, which was correlated with decreasing host plant biomass. Similarly, emergence proportions of parasitoids decreased with increasing plant species richness. Both, emergence proportions and proportions of female parasitoids were lower in plots with legumes, where host plants had increased nitrogen concentrations. This effect of legume presence could indicate that aphids were better defended against parasitoids in high-nitrogen environments. Body mass of emerged individuals of the two most abundant primary parasitoid species was, however, higher in plots with legumes, suggesting that once parasitoids could overcome aphid defenses, they could profit from larger or more nutritious hosts. Our study demonstrates that cascading effects of plant species loss on higher trophic levels such as aphids, parasitoids and secondary parasitoids begin with changed life-history traits of these insects. Thus, life-history traits of organisms at higher trophic levels may be useful indicators of bottom-up effects of plant diversity on the biodiversity of consumers.


The influence of green coffee bean extract supplementation on blood glucose levels: A systematic review and dose-response meta-analysis of randomized controlled trials.
Studies regarding the influence of green coffee extract (GCE) on blood glucose levels are conflicting. Thus, we sought to conduct a meta-analysis and systematic review of all available randomized controlled trials (RCTs) to quantify the effects of GCE and CGA intervention on blood glucose and insulin levels. We performed systematic online searches in Scopus, Web of science, and PubMed databases, from inception to July 2019. Data were combined analyzed using a random effects model (Der Simonian-Laird method) and reported as weighted mean differences (WMD). Ten trials reported the influences of GCE on FBS and insulin and were subsequently entered into the meta-analysis. Combined results highlighted that FBS was significantly altered after GCE consumption (WMD: -1.791 mg/dl, 95% CI -3.404, -0.177), with no significant heterogeneity among the studies (I2 = 35.0%, p = .128). However, overall results demonstrated that GCE administration did not result in any significant alteration in insulin levels (WMD: -0.925 μU/ml, 95% CI:-1.915, 0.064), with significant heterogeneity found across studies (I2 = 87.9%). In sub-group analysis, insulin levels were significantly reduced when GCE was supplemented in dosages of ≥400 mg/day (WMD:-1.942 mg/dl, 95% CI:-1.184, -0.975; I2 = 0.0%). The results of present study support the use of GCE for the enhancement of blood glucose, while subgroup analysis highlighted significant improvements in insulin levels when GCE is supplemented in doses ≥400 mg/day.


Stony endocarp dimension and shape variation in prunus section prunus.
Identification of Prunus groups at subspecies or variety level is complicated by the wide range of variation and morphological transitional states. Knowledge of the degree of variability within and between species is a sine qua non for taxonomists. Here, a detailed study of endocarp dimension and shape variation for taxa of Prunus section Prunus is presented. The sample size necessary to obtain an estimation of the population mean with a precision of 5 % was determined by iteration. Two cases were considered: (1) the population represents an individual; and (2) the population represents a species. The intra-individual and intraspecific variation of Prunus endocarps was studied by analysing the coefficients of variance for dimension and shape parameters. Morphological variation among taxa was assessed using univariate statistics. The influence of the time of sampling and the level of hydration on endocarp dimensions and shape was examined by means of pairwise t-tests. In total, 14 endocarp characters were examined for five Eurasian plum taxa. All linear measurements and index values showed a low or normal variability on the individual and species level. In contrast, the parameter 'Vertical Asymmetry' had high coefficients of variance for one or more of the taxa studied. Of all dimension and shape parameters studied, only 'Triangle' differed significantly between mature endocarps of P. insititia sampled with a time difference of 1 month. The level of hydration affected endocarp dimensions and shape significantly. Index values and the parameters 'Perimeter', 'Area', 'Triangle', 'Ellipse', 'Circular' and 'Rectangular', based on sample sizes and coefficients of variance, were found to be most appropriate for further taxonomic analysis. However, use of one, single endocarp parameter is not satisfactory for discrimination between Eurasian plum taxa, mainly because of overlapping ranges. Before analysing dried endocarps, full hydration is recommended, as this restores the original dimensions and shape.


Transcriptional repression of PTEN in neural cells using CRISPR/dCas9 epigenetic editing.
After damage to the adult mammalian central nervous system (CNS), surviving neurons have limited capacity to regenerate and restore functional connectivity. Conditional genetic deletion of PTEN results in robust CNS axon regrowth, while PTEN repression with short hairpin RNA (shRNA) improves regeneration but to a lesser extent, likely due to suboptimal PTEN mRNA knockdown using this approach. Here we employed the CRISPR/dCas9 system to repress PTEN transcription in neural cells. We targeted the PTEN proximal promoter and 5' untranslated region with dCas9 fused to the repressor protein Krüppel-associated box (KRAB). dCas9-KRAB delivered in a lentiviral vector with one CRISPR guide RNA (gRNA) achieved potent and specific PTEN repression in human cell line models and neural cells derived from human iPSCs, and induced histone (H)3 methylation and deacetylation at the PTEN promoter. The dCas9-KRAB system outperformed a combination of four shRNAs targeting the PTEN transcript, a construct previously used in CNS injury models. The CRISPR system also worked more effectively than shRNAs for Pten repression in rat neural crest-derived PC-12 cells, and enhanced neurite outgrowth after nerve growth factor stimulation. PTEN silencing with CRISPR/dCas9 epigenetic editing may provide a new option for promoting axon regeneration and functional recovery after CNS trauma.


Selective induction and subcellular distribution of ACONITASE 3 reveal the importance of cytosolic citrate metabolism during lipid mobilization in Arabidopsis.
Arabidopsis thaliana has three genes that encode distinct aconitases (ACO), but little is known about the function of each isoenzyme during plant development. In newly emerged seedlings of Arabidopsis, transcript and protein levels for ACO3 were selectively induced to yield more than 80% of total aconitase activity. Characterization of knockout mutants for each of the three ACOs suggests a major role for only ACO3 in citrate metabolism. The aco3 mutant showed delayed early seedling growth, altered assimilation of [14C]acetate feeding and elevated citrate levels, which were nearly 4-fold greater than in wild-type, aco1 or aco2. However, both ACO1 and ACO2 are active in seedlings as shown by inhibition of aco3 growth by the toxin monofluoroacetate, and altered [14C]acetate assimilation and metabolite levels in aco1 and aco2. Relative levels of fumarate and malate differed between aco2 and aco3, indicating metabolically isolated pools of these metabolites in seedlings. Our inability to enrich ACO protein through mitochondria isolation, and the reduced cytosolic ACO activity of the iron-sulfur centre assembly mutant atm3-1, indicated a cytosolic localization of ACO3 in 3-day-old seedlings. Subsequently, we determined that more than 90% of ACO3 was cytosolic. We conclude that ACO3 is cytosolic in young seedlings and functions in citrate catabolism consistent with the operation of the classic glyoxylate and not direct catabolism of citrate within mitochondria.


Hypertrygliceridemia-induced Acute Pancreatitis Following Hyperlipidemic Abdominal Crisis.
Hypertriglyceridemia is a well established cause of acute pancreatitis (AP). Multiple mechanism are proposed to explain this phenomenon, but the exact mechanism is unknown. Clinical manifestations are similar to other forms of AP. Although amylase and lipase levels exclude the diagnosis of AP in normolipidemic patients, they may be normal in hypertriglyceridemia-induced AP. Further evaluation with imaging studies are needed for diagnosis. A less known entity "hyperlipidemic abdominal crisis" is a prior clinical state before development to AP. We describe a young male patient without any previously diagnosed metabolic disorder presenting to emergency department with abdominal pain and vomiting where normal amylase in lactescent serum was detected. His abdomen computed tomography (CT) was reported as normal. His symptoms were relieved with antiemetic and histamine-2 blocker and he was diagnosed with hypertriglyceridemia and dyspepsia. Readmission with recurrence of symptoms after 3 hours resulted in hospitalization where the second CT showed edematous AP.


Increasing cardiopulmonary emergency visits by long-range transported Asian dust storms in Taiwan.
This study aims to explore whether Asian dust storms can affect health after 4000 km long-range transport from their origins to downwind areas. Asian dust storms reaching Taipei, Taiwan are tracked by satellite images and confirmed by backward trajectory analysis and ground air pollution monitoring between 1995 and 2002. Our outcome variables include emergency visits for ischaemic heart diseases (ICD-9-CM 410-411, 414), cerebrovascular diseases (ICD-9-CM 430-437), and chronic obstructive pulmonary diseases (COPD) (ICD-9-CM 493, 496) from the National Taiwan University Hospital (NTUH). We use simple paired t-test and Poisson regression models to compare difference in emergency visits, air pollution levels and meteorological conditions for the pairs of Asian dust events and pre-dust periods. There were 39 high dust events with PM(10) greater than 90 microg/m(3) and another 46 low dust events with PM(10) less than 90 microg/m(3). Compared to their pre-dust periods, PM(10) concentrations are significantly increased by 77 microg/m(3) per event for the high dust events. Asian dust storms increase cardiopulmonary emergency visits during storm-affecting periods in Taipei when ambient PM(10) concentrations are above 90 microg/m(3). Compared to their pre-dust periods, emergency visits for ischaemic heart diseases, cerebrovascular diseases, and COPD during high dust events are increased by 0.7 case (35%), 0.7 case (20%), and 0.9 case (20%) per event, respectively, by paired t-tests. By comparing the model-predicted to the observed emergency visits, we find emergency visits for cardiovascular diseases (ICD-9-CM 410-411, 414, 430-437) were significantly increased by 2.9 cases (67%) per event for the 39 high Asian dust events.


Efficacy of amitriptyline for treatment of somatoform pain disorder in the orofacial region: A case series.
To determine the efficacy of amitriptyline and the optimal dosage for treating a somatoform pain disorder in the orofacial region. Thirty outpatients with orofacial pain who fulfilled the criteria of pain disorder were recruited for the study. Twenty-three patients had specific precipitating events in their past history, which they considered to be the origin of the pain. Amitriptyline was administered and the dose was gradually increased up to a daily dose of 250 mg. The response to treatment was evaluated using a numeric rating scale and the Clinical Global Impression Scales. Five patients dropped out and 25 patients (83%) completed the trial. Twenty-two patients became pain free or nearly pain free, while 3 patients who also completed the study did not respond at all, even though they took a daily dose of 250 mg amitriptyline. For responders, the mean daily dose of amitriptyline was 77.5 +/- 51.5 mg (range, 10 to 200 mg). Four patients (16%) obtained pain relief with a daily dose of less than 50 mg, while 3 patients (12%) needed a daily dose of 150 mg or more for pain relief. Adverse side effects were observed in 19 patients. Amitriptyline was effective in relieving pain associated with a somatoform pain disorder in the orofacial region. The dose of amitriptyline may need to be as high as that used to treat a major depression.


Rare earth elements in freshwater, marine, and terrestrial ecosystems in the eastern Canadian Arctic.
Few ecotoxicological studies exist for rare earth elements (REEs), particularly field-based studies on their bioaccumulation and food web dynamics. REE mining has led to significant environmental impacts in several countries (China, Brazil, U.S.), yet little is known about the fate and transport of these contaminants of emerging concern. Northern ecosystems are potentially vulnerable to REE enrichment from prospective mining projects at high latitudes. To understand how REEs behave in remote northern food webs, we measured REE concentrations and carbon and nitrogen stable isotope ratios (∂15N, ∂13C) in biota from marine, freshwater, and terrestrial ecosystems of the eastern Canadian Arctic (N = 339). Wildlife harvesting and tissue sampling was partly conducted by local hunters through a community-based monitoring project. Results show that REEs generally follow a coherent bioaccumulation pattern for sample tissues, with some anomalies for redox-sensitive elements (Ce, Eu). Highest REE concentrations were found at low trophic levels, especially in vegetation and aquatic invertebrates. Terrestrial herbivores, ringed seal, and fish had low total REE levels in muscle tissue (∑REE for 15 elements <0.1 nmol g-1), yet accumulation was an order of magnitude higher in liver tissues. Age- and length-dependent REE accumulation also suggest that REE uptake is faster than elimination for some species. Overall, REE bioaccumulation patterns appear to be species- and tissue-specific, with limited potential for biomagnification. This study provides novel data on the behaviour of REEs in ecosystems and will be useful for environmental impact assessment of REE enrichment in northern regions.


Hypocretin antagonists in insomnia treatment and beyond.
Hypocretin neuropeptides have been shown to regulate transitions between wakefulness and sleep through stabilization of sleep promoting GABAergic and wake promoting cholinergic/monoaminergic neural pathways. Hypocretin also influences other physiologic processes such as metabolism, appetite, learning and memory, reward and addiction, and ventilatory drive. The discovery of hypocretin and its effect upon the sleep-wake cycle has led to the development of a new class of pharmacologic agents that antagonize the physiologic effects of hypocretin (i.e. hypocretin antagonists). Further investigation of these agents may lead to novel therapies for insomnia without the side-effect profile of currently available hypnotics (e.g. impaired cognition, confusional arousals, and motor balance difficulties). However, antagonizing a system that regulates the sleep-wake cycle while also influencing non-sleep physiologic processes may create an entirely different but equally concerning side-effect profile such as transient loss of muscle tone (i.e. cataplexy) and a dampened respiratory drive. In this review, we will discuss the discovery of hypocretin and its receptors, hypocretin and the sleep-wake cycle, hypocretin antagonists in the treatment of insomnia, and other implicated functions of the hypocretin system.


Molecular characterization of the humoral responses to Cryptococcus neoformans infection and glucuronoxylomannan-tetanus toxoid conjugate immunization.
The molecular characteristics of the humoral immune response to a serotype A Cryptococcus neoformans infection were compared with the response elicited by a cryptococcal glucuronoxylomannan-tetanus toxoid (GXM-TT) conjugate. Anticryptococcal monoclonal antibodies (mAbs) isolated from both responses have previously been shown to recognize the same antigenic determinant of cryptococcal GXM. Southern blot and sequence analyses indicate that the hybridomas isolated from each response arose from only a few precursor B cells. All the mAbs generated from the infected and GXM-TT conjugate-immunized mice utilize the same VH7183 family member: JH2/JH4, v kappa 5.1, and J kappa 1; mAbs generated by different B cells had complementarity-determining region 3's (CDR3s) composed of seven amino acids with a common sequence motif. Thus, the molecular analysis of these anticryptococcal mAb-producing hybridomas indicated that the response to both cryptococcal infection and conjugate immunization was oligoclonal and highly restricted with regard to immunoglobulin gene utilization. The GXM-TT conjugate primarily stimulated isotype switching and clonal proliferation, and did not result in hybridomas expressing additional immunoglobulin repertoires. The mAbs from both responses had a number of replacement mutations at the 5' end of CDR2 that appear to be the result of antigen-driven selection. Somatic mutation also resulted in altered epitope specificity for one mAb, 13F1. Passive administration of representative mAbs from different clones generated in response to the GXM-TT conjugate prolonged survival of lethally infected mice.


Mutations in AKAP5 disrupt dendritic signaling complexes and lead to electrophysiological and behavioral phenotypes in mice.
AKAP5 (also referred to as AKAP150 in rodents and AKAP79 in humans) is a scaffolding protein that is highly expressed in neurons and targets a variety of signaling molecules to dendritic membranes. AKAP5 interacts with PKA holoenzymes containing RIIalpha or RIIbeta as well as calcineurin (PP2B), PKC, calmodulin, adenylyl cyclase type V/VI, L-type calcium channels, and beta-adrenergic receptors. AKAP5 has also been shown to interact with members of the MAGUK family of PSD-scaffolding proteins including PSD95 and SAP97 and target signaling molecules to receptors and ion channels in the postsynaptic density (PSD). We created two lines of AKAP5 mutant mice: a knockout of AKAP5 (KO) and a mutant that lacks the PKA binding domain of AKAP5 (D36). We find that PKA is delocalized in both the hippocampus and striatum of KO and D36 mice indicating that other neural AKAPs cannot compensate for the loss of PKA binding to AKAP5. In AKAP5 mutant mice, a significant fraction of PKA becomes localized to dendritic shafts and this correlates with increased binding to microtubule associated protein-2 (MAP2). Electrophysiological and behavioral analysis demonstrated more severe deficits in both synaptic plasticity and operant learning in the D36 mice compared with the complete KO animals. Our results indicate that the targeting of calcineurin or other binding partners of AKAP5 in the absence of the balancing kinase, PKA, leads to a disruption of synaptic plasticity and results in learning and memory defects.


[The effect of alternating administration of aluminum chloride and sodium fluoride in drinking water on the concentration of fluoride in serum and its content in bones of rats].
Fluorine and aluminum remain a very interesting research topic due to equivocal and relatively unknown toxic action, role in the etiology of various diseases, and interactions of both elements. Fluorine and aluminum compounds are absorbed by organisms through the gastric and respiratory systems, although the latter route operates only at very high concentrations in air. Chronic exposure to fluorine and aluminum leads to accumulation of both elements, especially in bones and teeth, but also in lung, brain, kidney, and liver. Organisms excrete these elements with urine, faeces, and to a minor extent with sweat and bile. In the light of reports suggesting that aluminum has protective properties against fluorine toxicity during exposure to both elements, we decided to examine the effect of alternating doses of aluminum fluoride and sodium fluoride in drinking water on rats. Four female groups received: I--100 ppm fluorine ions during one month; II--100 ppm fluorine ions alternating every two days with 300 ppm aluminum ions during one month; III--100 ppm fluoride ions during four months; IV--100 ppm fluorine ions alternating every two days with 300 ppm aluminum ions during four months. The respective male groups called IA, IIA, IIIA, and IVA were treated identically. Subsequently, the animals were anesthetized and sacrificed. Blood was sampled from the heart and the right femur was removed for fluorine determination. Fluorine content in the femur and serum was determined with an ion-selective electrode (Orion). The results were analyzed statistically (Statistica 6). We observed higher fluoride concentrations in serum as compared with control values in all groups of female and male rats exposed to sodium fluoride only. Longer exposure time (4 months) did not result in further increase in serum fluoride concentration. However, longer exposure increased fluoride accumulation in the femur (p < 0.001). All groups exposed to NaF had significantly higher fluoride concentration in the femur as compared with control animals. Groups receiving NaF and AlCl3 showed lower fluoride concentration in serum and femur compared with those exposed to NaF only and higher in comparison with controls. Fluorine content in the femur of rats exposed to NaF and AlCI3 for four months was similar to the results obtained after one month of exposure.


Eccrine Porocarcinoma: A Multicenter Retrospective Study with Review of the Literatures Reported in Korea.
Eccrine porocarcinoma (EPC) is a rare malignant cutaneous adnexal tumor. Other than several scattered case reports, no comprehensive review on EPC has been conducted in Korea. To clinicopathologically review all EPC cases from our institutions as well as those reported in Korea. Medical records and histopathological slides of EPC cases in the skin biopsy registries of our institutions were retrospectively reviewed. Additionally, EPC cases reported in Korea before June 2019 were retrieved by searching the PubMed, KoMCI, KoreaMed, and KMbase databases. Nine EPC cases from our institutions were included in the study. In addition, 27 reports of 28 patients with EPC were reported in Korea. A total of 37 patients with EPC were identified, consisting of 19 males (male:female ratio, 1.06:1; mean age at diagnosis, 65.6 years). The most common site of primary tumor was the head and neck (29.7%). Wide excision was the most common (78.4%) treatment method. Initial metastasis work-up imaging studies were performed in 18 patients (48.6%), and metastasis was confirmed in eight patients (21.6%). EPC is a rare cutaneous carcinoma in Korea. EPC usually affects elderly patients, with no sexual predilection. Due to possible metastasis, careful diagnosis and appropriate metastasis work-ups are warranted in EPC.


Normal scores of deep breathing tests: beware of dysrhythmia in transthyretin amyloidosis.
The heart rate (HR) response to paced deep breathing (DB) is a common test of cardiac autonomic function, where high heart rate variability (HRV) is considered to reflect normal autonomic function. We evaluated the DB test in patients with hereditary transthyretin amyloid (ATTRm) amyloidosis, where autonomic dysregulation and atrial arrhythmias are common. Paced DB was performed during one minute (six breaths/min) in 165 recordings in adult ATTRm amyloidosis patients with the TTR Val30Met mutation, 42 hypertrophic cardiomyopathy (HCM) patients and 211 healthy subjects. HRV was scored by traditional DB indices and by a novel regularity index, estimating the fraction of the HRV that was coherent with the breathing pattern. Twenty per cent of ATTRm amyloidosis patients presented with age-adjusted HRV scores within normal limits but poor regularity due to subtle atrial arrhythmias and cardiac conduction disturbances. Forty-seven per cent of ATTRm amyloidosis patients presented with HRV scores below normal limits, whereas HCM patients presented with higher HRV than ATTRm amyloidosis patients. Reduced HRV is common in ATTRm amyloidosis patients during DB, however, autonomic function cannot be evaluated in patients presenting with the combination of "normal" scores and low regularity, since their HR responses often reflects dysrhythmias.


Recurrent DUX4 fusions in B cell acute lymphoblastic leukemia of adolescents and young adults.
The oncogenic mechanisms underlying acute lymphoblastic leukemia (ALL) in adolescents and young adults (AYA; 15-39 years old) remain largely elusive. Here we have searched for new oncogenes in AYA-ALL by performing RNA-seq analysis of Philadelphia chromosome (Ph)-negative AYA-ALL specimens (n = 73) with the use of a next-generation sequencer. Interestingly, insertion of D4Z4 repeats containing the DUX4 gene into the IGH locus was frequently identified in B cell AYA-ALL, leading to a high level of expression of DUX4 protein with an aberrant C terminus. A transplantation assay in mice demonstrated that expression of DUX4-IGH in pro-B cells was capable of generating B cell leukemia in vivo. DUX4 fusions were preferentially detected in the AYA generation. Our data thus show that DUX4 can become an oncogenic driver as a result of somatic chromosomal rearrangements and that AYA-ALL may be a clinical entity distinct from ALL at other ages.


Phylogeny of whey acidic protein (WAP) four-disulfide core proteins and their role in lower vertebrates and invertebrates.
Proteins containing WAP (whey acidic protein) domains with a characteristic WFDC (WAP four-disulfide core) occur not only in mammals (including marsupials and monotremes) but also in birds, reptiles, amphibians and fish. In addition, they are present in numerous invertebrates, from cnidarians to urochordates. Many of those from non-mammalian groups are poorly understood with respect to function or phylogeny. Those well characterized so far are waprins from snakes, perlwapins from bivalves and crustins from decapod crustaceans. Waprins are venom proteins with a single WAP domain at the C-terminus. They display antimicrobial, rather than proteinase inhibitory, activities. Perlwapins, in contrast, possess three WAP domains at the C-terminus and are expressed in the shell nacre of abalones. They participate in shell formation by inhibiting the growth of calcium crystals in the shell. The crustin group is the largest of all WFDC-containing proteins in invertebrates with the vast majority being highly expressed in the haemocytes. Most have a single WAP domain at the C-terminus. The presence and type of the domains between the signal sequence and the C-terminus WAP domain separate the different crustin types. Most of the Type I and II crustins are antimicrobial towards Gram-positive bacteria, whereas the Type III crustins tend to display protease inhibition. Expression studies show that at least some crustins have other important biological effects, as levels change with physiological stress, wound repair, tissue regeneration or ecdysis. Thus WAP domains are widely distributed and highly conserved, serving in diverse physiological processes (proteinase inhibition, bacterial killing or inhibition of calcium transport).


Functional magnetic resonance imaging of reorganization in rat brain after stroke.
Functional recovery after stroke has been associated with brain plasticity; however, the exact relationship is unknown. We performed behavioral tests, functional MRI, and histology in a rat stroke model to assess the correlation between temporal changes in sensorimotor function, brain activation patterns, cerebral ischemic damage, and cerebrovascular reactivity. Unilateral stroke induced a large ipsilateral infarct and acute dysfunction of the contralateral forelimb, which significantly recovered at later stages. Forelimb impairment was accompanied by loss of stimulus-induced activation in the ipsilesional sensorimotor cortex; however, local tissue and perfusion were only moderately affected and cerebrovascular reactivity was preserved in this area. At 3 days after stroke, extensive activation-induced responses were detected in the contralesional hemisphere. After 14 days, we found reduced involvement of the contralesional hemisphere, and significant responses in the infarction periphery. Our data suggest that limb dysfunction is related to loss of brain activation in the ipsilesional sensorimotor cortex and that restoration of function is associated with biphasic recruitment of peri- and contralesional functional fields in the brain.