Emerging investigator series: dual role of organic matter in the anaerobic degradation of triclosan.
Triclosan (TCS), one of the most widely used antimicrobial agents, has been listed among the top 10 contaminants in US rivers. Environmental persistence, endocrine disruption effects, and the antibiotic resistance induction capacity of TCS attract interest in its environmental fate and degradation. Herein, we found that TCS can be anaerobically degraded at pH 9 by a metal-reducing bacterium, Shewanella putrefaciens CN32. The degradation was substantially facilitated by low-concentration (0-15 mg C per L) organic matter (OM) extracted from a peat soil, whereas TCS degradation was inhibited by further increased concentration (15-100 mg C per L) of OM. OM acted as both an electron shuttle and sorbent in regulating the degradation of TCS. The novel dual role of ubiquitous OM in the reaction of TCS governs the environmental degradation and persistence of TCS. Our study highlights the effects of OM on the reaction of emerging trace organic pollutants, with implications on their engineering treatment and environmental risk regulation.


Bradykinin, angiotensin-(1-7), and ACE inhibitors: how do they interact?
The beneficial effect of ACE inhibitors in hypertension and heart failure may relate, at least in part, to their capacity to interfere with bradykinin metabolism. In addition, recent studies have provided evidence for bradykinin-potentiating effects of ACE inhibitors that are independent of bradykinin hydrolysis, i.e. ACE-bradykinin type 2 (B(2)) receptor 'cross-talk', resulting in B(2) receptor upregulation and/or more efficient activation of signal transduction pathways, as well as direct activation of bradykinin type 1 receptors by ACE inhibitors. This review critically reviews the current evidence for hydrolysis-independent bradykinin potentiation by ACE inhibitors, evaluating not only the many studies that have been performed with ACE-resistant bradykinin analogues, but also paying attention to angiotensin-(1-7), a metabolite of both angiotensin I and II, that could act as an endogenous ACE inhibitor. The levels of angiotensin-(1-7) are increased during ACE inhibition, and most studies suggest that its hypotensive effects are mediated in a bradykinin-dependent manner.


POLARIZED SPATIAL FREQUENCY DOMAIN IMAGING OF HEART VALVE FIBER STRUCTURE.
Our group previously introduced Polarized Spatial Frequency Domain Imaging (PSFDI), a wide-field, reflectance imaging technique which we used to empirically map fiber direction in porcine pulmonary heart valve leaflets (PHVL) without optical clearing or physical sectioning of the sample. Presented is an extended analysis of our PSFDI results using an inverse Mueller matrix model of polarized light scattering that allows additional maps of fiber orientation distribution, along with instrumentation permitting increased imaging speed for dynamic PHVL fiber measurements. We imaged electrospun fiber phantoms with PSFDI, and then compared these measurements to SEM data collected for the same phantoms. PHVL was then imaged and compared to results of the same leaflets optically cleared and imaged with small angle light scattering (SALS). The static PHVL images showed distinct regional variance of fiber orientation distribution, matching our SALS results. We used our improved imaging speed to observe bovine tendon subjected to dynamic loading using a biaxial stretching device. Our dynamic imaging experiment showed trackable changes in the fiber microstructure of biological tissue under loading. Our new PSFDI analysis model and instrumentation allows characterization of fiber structure within heart valve tissues (as validated with SALS measurements), along with imaging of dynamic fiber remodeling. The experimental data will be used as inputs to our constitutive models of PHVL tissue to fully characterize these tissues' elastic behavior, and has immediate application in determining the mechanisms of structural and functional failure in PHVLs used as bio-prosthetic implants.


Farmers and Climate Change: A Cross-National Comparison of Beliefs and Risk Perceptions in High-Income Countries.
Climate change has serious implications for the agricultural industry-both in terms of the need to adapt to a changing climate and to modify practices to mitigate for the impacts of climate change. In high-income countries where farming tends to be very intensive and large scale, it is important to understand farmers' beliefs and concerns about climate change in order to develop appropriate policies and communication strategies. Looking across six study sites-Scotland, Midwestern United States, California, Australia, and two locations in New Zealand-this paper finds that over half of farmers in each location believe that climate change is occurring. However, there is a wide range of beliefs regarding the anthropogenic nature of climate change; only in Australia do a majority of farmers believe that climate change is anthropogenic. In all locations, a majority of farmers believe that climate change is not a threat to local agriculture. The different policy contexts and existing impacts from climate change are discussed as possible reasons for the variation in beliefs. This study compared varying surveys from the different locations and concludes that survey research on farmers and climate change in diverse locations should strive to include common questions to facilitate comparisons.


Personality traits of centenarians' offspring.
To determine whether the offspring of centenarians have personality characteristics that are distinct from the general population. Case-control. Nationwide U.S. sample. Unrelated offspring of centenarians (n=246, mean age 75) were compared with published norms. Using the NEO-Five-Factor Inventory (NEO-FFI) questionnaire, measures of the personality traits neuroticism, extraversion, openness, agreeableness, and conscientiousness were obtained. T-scores and percentiles were calculated according to sex and used to interpret the results. Male and female offspring of centenarians scored in the low range of published norms for neuroticism and in the high range for extraversion. The women also scored comparatively high in agreeableness. Otherwise, both sexes scored within normal range for conscientiousness and openness, and the men scored within normal range for agreeableness. Specific personality traits may be important to the relative successful aging demonstrated by the offspring of centenarians. Similarities across four of the five domains between male and female offspring is noteworthy and may relate to their successful aging. Measures of personality are an important phenotype to include in studies that assess genetic and environmental influences of longevity and successful aging.


Pulmonary infections complicating ARDS.
Pulmonary infection is one of the main complications occurring in patients suffering from acute respiratory distress syndrome (ARDS). Besides traditional risk factors, dysregulation of lung immune defenses and microbiota may play an important role in ARDS patients. Prone positioning does not seem to be associated with a higher risk of pulmonary infection. Although bacteria associated with ventilator-associated pneumonia (VAP) in ARDS patients are similar to those in patients without ARDS, atypical pathogens (Aspergillus, herpes simplex virus and cytomegalovirus) may also be responsible for infection in ARDS patients. Diagnosing pulmonary infection in ARDS patients is challenging, and requires a combination of clinical, biological and microbiological criteria. The role of modern tools (e.g., molecular methods, metagenomic sequencing, etc.) remains to be evaluated in this setting. One of the challenges of antimicrobial treatment is antibiotics diffusion into the lungs. Although targeted delivery of antibiotics using nebulization may be interesting, their place in ARDS patients remains to be explored. The use of extracorporeal membrane oxygenation in the most severe patients is associated with a high rate of infection and raises several challenges, diagnostic issues and pharmacokinetics/pharmacodynamics changes being at the top. Prevention of pulmonary infection is a key issue in ARDS patients, but there is no specific measure for these high-risk patients. Reinforcing preventive measures using bundles seems to be the best option.


[Risk of postoperative infection in patients with inflammatory bowel disease].
The clinical course of patients with inflammatory bowel disease (IBD) frequently leads to the use of immunosuppressants and immunomodulators. We investigated the risk of postoperative infection in patients with IBD undergoing elective bowel surgery and whether the use of corticosteroid (CS) and/or 6-mercaptopurine/ azathioprine (6-MP/AZA) before surgery was associated with the increased risk of postoperative infection. Patients who were diagnosed as Crohn's disease (n=25) or ulcerative colitis (n=19) and underwent elective bowel surgery between 1986 and 2005 were identified. Medical records were retrospectively analyzed including age, sex, duration of disease, indication for surgery, duration of surgery, type of surgery, type of postoperative infection, admission period, usage of CS and 6-MP/AZA, and preoperative laboratory values. There were 27 patients receiving CS alone, 6 patients receiving 6-MP/AZA alone or with CS, and 16 patients receiving neither CS nor 6-MP/AZA. There were 17 postoperative infections (38.6%) among IBD patients who had undergone surgery and wound infection was the most common type of infection (76.5%). In IBD patients, patients receiving CS had higher postoperative infection rate than those patients receiving neither CS nor 6-MP/AZA (p=0.039). Patients receiving CS in conjunction with 6-MP/AZA did not have significantly higher postoperative infection rate than those with CS only (p=0.415). Preoperative use of CS in patients with IBD is associated with the increased risk of postoperative infections. Addition of 6-MP/AZA in patients receiving CS does not increase the risk of postoperative infections.


Feasibility of cardiac gating free of interference with electro-magnetic fields at 1.5 Tesla, 3.0 Tesla and 7.0 Tesla using an MR-stethoscope.
To circumvent the challenges of conventional electrocardiographic (ECG)-gating by examining the efficacy of an MR stethoscope, which offers (i) no risk of high voltage induction or patient burns, (ii) immunity to electromagnetic interference, (iii) suitability for all magnetic field strengths, and (iv) patient comfort together with ease of use for the pursuit of reliable and safe (ultra)high field cardiac gated magnetic resonance imaging (MRI). The acoustic gating device consists of 3 main components: an acoustic sensor, a signal processing unit, and a coupler unit to the MRI system. Signal conditioning and conversion are conducted outside the 0.5 mT line using dedicated electronic circuits. The final waveform is delivered to the internal physiological signal controller circuitry of a clinical MR scanner. Cardiovascular MRI was performed of normal volunteers (n = 17) on 1.5 T, 3.0 T and 7.0 T whole body MR systems. Black blood imaging, 2D CINE imaging, 3D phase contrast MR angiography, and myocardial T2* mapping were carried out. The MR-stethoscope provided cardiograms at 1.5 T, 3.0 T and 7.0 T free of interference from electromagnetic fields and magneto-hydrodynamic effects. In comparison, ECG waveforms were susceptible to T-wave elevation and other distortions, which were more pronounced at higher fields. Acoustically gated black blood imaging at 1.5 T and 3.0 T provided image quality comparable with or even superior to that obtained from the ECG-gated approach. In the case of correct R-wave recognition, ECG-gated 2D CINE SSFP imaging was found to be immune to cardiac motion effects -even at 3.0 T. However, ECG-gated 2D SSFP CINE imaging was prone to cardiac motion artifacts if R-wave mis-registration occurred because of T-wave elevation. Acoustically gated 3D PCMRA at 1.5 T, 3.0 T and 7.0 T resulted in images free of blood pulsation artifacts because the acoustic gating approach provided cardiac signal traces free of interference with electromagnetic fields or magneto-hydrodynamic effects even at 7.0 Tesla. Severe ECG-trace distortions and T-wave elevations occurred at 3.0 T and 7.0 T. Acoustically cardiac gated T2* mapping at 3.0 T yielded a T2* value of 22.3 +/- 4.8 ms for the inferoseptal myocardium. The proposed MR-stethoscope presents a promising alternative to currently available techniques for cardiac gating of (ultra)high field MRI. Its intrinsic insensitivity to interference from electromagnetic fields renders it suitable for clinical imaging because of its excellent trigger reliability, even at 7.0 Tesla.


Medical school curriculum characteristics associated with intentions and frequency of tobacco dependence treatment among 3rd year U.S. medical students.
Physicians play a critical role in addressing tobacco dependence, yet report limited training. Tobacco dependence treatment curricula for medical students could improve performance in this area. This study identified student and medical school tobacco treatment curricula characteristics associated with intentions and use of the 5As for tobacco treatment among 3rd year U.S. medical students. Third year medical students (N=1065, 49.3% male) from 10 U.S. medical schools completed a survey in 2009-2010 assessing student characteristics, including demographics, tobacco treatment knowledge, and self-efficacy. Tobacco curricula characteristics assessed included amount and type of classroom instruction, frequency of tobacco treatment observation, instruction, and perception of preceptors as role models. Greater tobacco treatment knowledge, self-efficacy, and curriculum-specific variables were associated with 5A intentions, while younger age, tobacco treatment self-efficacy, intentions, and each curriculum-specific variable were associated with greater 5A behaviors. When controlling for important student variables, greater frequency of receiving 5A instruction (OR=1.07; 95%CI 1.01-1.12) and perception of preceptors as excellent role models in tobacco treatment (OR=1.35; 95%CI 1.04-1.75) were significant curriculum predictors of 5A intentions. Greater 5A instruction (B=.06 (.03); p<.05) and observation of tobacco treatment (B=.35 (.02); p<.001) were significant curriculum predictors of greater 5A behaviors. Greater exposure to tobacco treatment teaching during medical school is associated with both greater intentions to use and practice tobacco 5As. Clerkship preceptors, or those physicians who provide training to medical students, may be particularly influential when they personally model and instruct students in tobacco dependence treatment.


Application of three-dimensional rendering in joint-related ganglion cysts.
The origin of para-articular cysts is poorly understood and controversial. The relatively common, simple (extraneural) cysts are presumed to be derived from joints, although joint connections are not always established. Rarer complex cysts are thought by many to form de novo within nerves (intraneural ganglion cysts) or within vessels (adventitial cysts) (degenerative theory). We believe that these simple and complex ganglion cysts are joint-related (articular theory). Joint connections are often not readily appreciated with routine imaging or at surgery. Not identifying and/or treating joint connections frequently leads to cyst recurrence. More sophisticated imaging may enhance visualization of these joint connections. We created a 3D rendering technique to assess potential joint connections of simple and complex cysts localized to the knee and superior tibiofibular joints in patients with fibular (peroneal) neuropathy. Two- and three-dimensional data sets from MRI examinations were segmented semiautomatically by signal intensity with further refinement based on interaction with the user to identify specific anatomic structures, such as small nerves and vessels on serial images. The bone, cysts, nerves, and vessels were each assigned different color representations, and 3D renderings were created in ANALYZE using the data sets closest to isotropic (voxel with equal length in all dimensions) resolution as the primary background rendering. We selected four cases to illustrate the spectrum of pathology. In all of these cases, we demonstrated joint connections and correlated imaging and operative findings. Surgery addressing the cyst and the joint connection resulted in excellent outcomes; postoperative MRIs done more than 6 months later confirmed that there was no recurrence. In addition to highlighting the important relationship of these cysts to neighboring anatomic structures, this 3D technique allows visualization of "occult" connections not readily appreciated with standard MR imaging. We believe that these joint-related cysts have a common pathogenesis; they dissect through a capsular rent and follow the path of least resistance; they may form simple cysts by dissecting out into the soft tissue, or more complex cysts by dissecting within the epineurium of nerves or adventitia of vessels (along an articular branch), or various combinations of all of these types of cysts. Understanding the pathogenesis for cyst formation will improve surgical management and outcomes. We have adapted this 3D technique to enhance the visualization of cysts occurring at other joints.


Characterizing poliovirus transmission and evolution: insights from modeling experiences with wild and vaccine-related polioviruses.
With national and global health policymakers facing numerous complex decisions related to achieving and maintaining polio eradication, we expanded our previously developed dynamic poliovirus transmission model using information from an expert literature review process and including additional immunity states and the evolution of oral poliovirus vaccine (OPV). The model explicitly considers serotype differences and distinguishes fecal-oral and oropharyngeal transmission. We evaluated the model by simulating diverse historical experiences with polioviruses, including one country that eliminated wild poliovirus using both OPV and inactivated poliovirus vaccine (IPV) (USA), three importation outbreaks of wild poliovirus (Albania, the Netherlands, Tajikistan), one situation in which no circulating vaccine-derived polioviruses (cVDPVs) emerge despite annual OPV use and cessation (Cuba), three cVDPV outbreaks (Haiti, Madura Island in Indonesia, northern Nigeria), one area of current endemic circulation of all three serotypes (northern Nigeria), and one area with recent endemic circulation and subsequent elimination of multiple serotypes (northern India). We find that when sufficient information about the conditions exists, the model can reproduce the general behavior of poliovirus transmission and outbreaks while maintaining consistency in the generic model inputs. The assumption of spatially homogeneous mixing remains a significant limitation that affects the performance of the differential equation-based model when significant heterogeneities in immunity and mixing may exist. Further studies on OPV virus evolution and improved understanding of the mechanisms of mixing and transmission may help to better characterize poliovirus transmission in populations. Broad application of the model promises to offer insights in the context of global and national policy and economic models.


Autoimmune disease classification by inverse association with SNP alleles.
With multiple genome-wide association studies (GWAS) performed across autoimmune diseases, there is a great opportunity to study the homogeneity of genetic architectures across autoimmune disease. Previous approaches have been limited in the scope of their analysis and have failed to properly incorporate the direction of allele-specific disease associations for SNPs. In this work, we refine the notion of a genetic variation profile for a given disease to capture strength of association with multiple SNPs in an allele-specific fashion. We apply this method to compare genetic variation profiles of six autoimmune diseases: multiple sclerosis (MS), ankylosing spondylitis (AS), autoimmune thyroid disease (ATD), rheumatoid arthritis (RA), Crohn's disease (CD), and type 1 diabetes (T1D), as well as five non-autoimmune diseases. We quantify pair-wise relationships between these diseases and find two broad clusters of autoimmune disease where SNPs that make an individual susceptible to one class of autoimmune disease also protect from diseases in the other autoimmune class. We find that RA and AS form one such class, and MS and ATD another. We identify specific SNPs and genes with opposite risk profiles for these two classes. We furthermore explore individual SNPs that play an important role in defining similarities and differences between disease pairs. We present a novel, systematic, cross-platform approach to identify allele-specific relationships between disease pairs based on genetic variation as well as the individual SNPs which drive the relationships. While recognizing similarities between diseases might lead to identifying novel treatment options, detecting differences between diseases previously thought to be similar may point to key novel disease-specific genes and pathways.


The phosphatidylcholine-transfer protein catalyzed import of phosphatidylcholine into isolated rat liver mitochondria.
In order to study the individual steps involved in the import of phosphatidylcholine (PC) into rat liver mitochondria, a number of PC analogues were introduced into the outer membrane of isolated mitochondria. Two fluorescent PC species, i.e. 1-palmitoyl-2-(16-bimanylthio)hexadecanoyl-PC (bimane-PC) and 1-palmitoyl-2-(10-pyrene)decanoyl-PC (pyrene-PC), and one radiolabeled PC species, i.e. 1-palmitoyl-2-[1-14C]oleoyl-PC (14C-POPC), were studied. The PC analogues were introduced from small unilamellar vesicles with the use of PC-specific transfer protein. The amount of PC imported was quantified by reisolation of the mitochondria. Import of the fluorescent PC species was monitored by on-line fluorescence spectroscopy. The distribution of the newly inserted PC between the outer and the inner membrane was assessed by separation of the two membranes using digitonin treatment. All analogues tested remained exclusively localized in the outer membrane thereby suggesting that additional (extramitochondrial) factors are required to initiate transfer of PC to the inner membrane.


A programmable platform for sub-second multichemical dynamic stimulation and neuronal functional imaging in C. elegans.
Caenorhabditis elegans (C. elegans) is a prominent model organism in neuroscience, as its small stereotyped nervous system offers unique advantages for studying neuronal circuits at the cellular level. Characterizing temporal dynamics of neuronal circuits is essential to fully understand neuronal processing. Characterization of the temporal dynamics of chemosensory circuits requires a precise and fast method to deliver multiple stimuli and monitor the animal's neuronal activity. Microfluidic platforms have been developed that offer an improved control of chemical delivery compared to manual methods. However, stimulating an animal with multiple chemicals at high speed is still difficult. In this work, we have developed a platform that can deliver any sequence of multiple chemical reagents, at sub-second resolution and without cross-contamination. We designed a network of chemical selectors wherein the chemical selected for stimulation is determined by the set of pressures applied to the chemical reservoirs. Modulation of inlet pressures has been automated to create robust, programmable sequences of subsecond chemical pulses. We showed that stimulation with sequences of different chemicals at the second to sub-second range can generate different neuronal activity patterns in chemosensory neurons; we observed previously unseen neuronal responses to a controlled chemical stimulation. Because of the speed and versatility of stimulus generated, this platform opens new possibilities to investigate neuronal circuits.


Dielectric and thermal effects on the optical properties of natural dyes: a case study on solvated cyanin.
The optical properties of the flavylium state of the cyanin dye are simulated numerically by combining Car-Parrinello molecular dynamics and linear-response time-dependent density functional theory calculations. The spectrum of the dye calculated in the gas phase is characterized by two peaks in the yellow and in the blue (green and violet), using a GGA-PBE (hybrid-B3LYP) DFT functional, which would bring about a greenish (bright orange) color incompatible with the dark purple hue observed in nature. Describing the effect of the water solvent through a polarizable continuum model does not modify qualitatively the resulting picture. An explicit simulation of both solvent and thermal effects using ab initio molecular dynamics results instead in a spectrum that is compatible with the observed coloration. This result is analyzed in terms of the spectroscopic effects of the molecular distortions induced by thermal fluctuations.


Direct 17O Isotopic Labeling of Oxides Using Mechanochemistry.
While 17O NMR is increasingly being used for elucidating the structure and reactivity of complex molecular and materials systems, much effort is still required for it to become a routine analytical technique. One of the main difficulties for its development comes from the very low natural abundance of 17O (0.04%), which implies that isotopic labeling is generally needed prior to NMR analyses. However, 17O-enrichment protocols are often unattractive in terms of cost, safety, and/or practicality, even for compounds as simple as metal oxides. Here, we demonstrate how mechanochemistry can be used in a highly efficient way for the direct 17O isotopic labeling of a variety of s-, p-, and d-block oxides, which are of major interest for the preparation of functional ceramics and glasses: Li2O, CaO, Al2O3, SiO2, TiO2, and ZrO2. For each oxide, the enrichment step was performed under ambient conditions in less than 1 h and at low cost, which makes these synthetic approaches highly appealing in comparison to the existing literature. Using high-resolution solid-state 17O NMR and dynamic nuclear polarization, atomic-level insight into the enrichment process is achieved, especially for titania and alumina. Indeed, it was possible to demonstrate that enriched oxygen sites are present not only at the surface but also within the oxide particles. Moreover, information on the actual reactions occurring during the milling step could be obtained by 17O NMR, in terms of both their kinetics and the nature of the reactive species. Finally, it was demonstrated how high-resolution 17O NMR can be used for studying the reactivity at the interfaces between different oxide particles during ball-milling, especially in cases when X-ray diffraction techniques are uninformative. More generally, such investigations will be useful not only for producing 17O-enriched precursors efficiently but also for understanding better mechanisms of mechanochemical processes themselves.


Anti-Metastatic and Anti-Invasion Effects of a Specific Anti-MUC18 scFv Antibody on Breast Cancer Cells.
Breast cancer is the most common malignancy in women. Altered expression of MUC18, a cell surface receptor, and its interaction with Wnt-5a as its ligand, affects the motility and invasiveness of breast cancer cells. In this study, we explored the Wnt-5a binding site and designed an antigenic epitope on the MUC18 receptor using in silico methods. A specific single-chain variable fragment (scFv) was isolated against the epitope by several panning processes. The binding ability of the scFv to the related epitope was evaluated in ELISA and flow cytometry. The inhibitory effects of the selected scFv on MUC18 positive cell line, MDA-MB231, was assessed by migration and invasion assays. The results demonstrated isolation of specific scFv with frequency of 40 % which showed significant binding with the epitope in both ELISA and fluorescence-activated cell sorting (FACS) analyses. The antibody inhibited the migration (76 %) and invasion (67 %) of MUC18 positive cell line. The results suggest the specific anti-MUC18 scFv as an effective antibody for breast cancer immunotherapy.


Multistep nucleation of nanocrystals in aqueous solution.
The nucleation and growth of solids from solutions impacts many natural processes and is fundamental to applications in materials engineering and medicine. For a crystalline solid, the nucleus is a nanoscale cluster of ordered atoms that forms through mechanisms still poorly understood. In particular, it is unclear whether a nucleus forms spontaneously from solution via a single- or multiple-step process. Here, using in situ electron microscopy, we show how gold and silver nanocrystals nucleate from supersaturated aqueous solutions in three distinct steps: spinodal decomposition into solute-rich and solute-poor liquid phases, nucleation of amorphous nanoclusters within the metal-rich liquid phase, followed by crystallization of these amorphous clusters. Our ab initio calculations on gold nucleation suggest that these steps might be associated with strong gold-gold atom coupling and water-mediated metastable gold complexes. The understanding of intermediate steps in nuclei formation has important implications for the formation and growth of both crystalline and amorphous materials.


Structure-based engineering of minimal proline dehydrogenase domains for inhibitor discovery.
Proline dehydrogenase (PRODH) catalyzes the FAD-dependent oxidation of l-proline to Δ1-pyrroline-5-carboxylate and is a target for inhibitor discovery because of its importance in cancer cell metabolism. Because human PRODH is challenging to purify, the PRODH domains of the bacterial bifunctional enzyme proline utilization A (PutA) have been used for inhibitor development. These systems have limitations due to large polypeptide chain length, conformational flexibility and the presence of domains unrelated to PRODH activity. Herein, we report the engineering of minimal PRODH domains for inhibitor discovery. The best designs contain one-third of the 1233-residue PutA from Sinorhizobium meliloti and include a linker that replaces the PutA α-domain. The minimal PRODHs exhibit near wild-type enzymatic activity and are susceptible to known inhibitors and inactivators. Crystal structures of minimal PRODHs inhibited by S-(-)-tetrahydro-2-furoic acid and 2-(furan-2-yl)acetic acid were determined at 1.23 and 1.72 Å resolution. Minimal PRODHs should be useful in chemical probe discovery.


Controlling measles using supplemental immunization activities: a mathematical model to inform optimal policy.
The Measles & Rubella Initiative, a broad consortium of global health agencies, has provided support to measles-burdened countries, focusing on sustaining high coverage of routine immunization of children and supplementing it with a second dose opportunity for measles vaccine through supplemental immunization activities (SIAs). We estimate optimal scheduling of SIAs in countries with the highest measles burden. We develop an age-stratified dynamic compartmental model of measles transmission. We explore the frequency of SIAs in order to achieve measles control in selected countries and two Indian states with high measles burden. Specifically, we compute the maximum allowable time period between two consecutive SIAs to achieve measles control. Our analysis indicates that a single SIA will not control measles transmission in any of the countries with high measles burden. However, regular SIAs at high coverage levels are a viable strategy to prevent measles outbreaks. The periodicity of SIAs differs between countries and even within a single country, and is determined by population demographics and existing routine immunization coverage. Our analysis can guide country policymakers deciding on the optimal scheduling of SIA campaigns and the best combination of routine and SIA vaccination to control measles.


Cloning and sequence analysis of rat fertilin alpha and beta--developmental expression, processing and immunolocalization.
Fertilin alpha and beta are members of the MDC (metalloproteinase-like, disintegrin-like, cysteine-rich) protein family and are expressed on the sperm surface where they have been proposed to play a role in mammalian fertilization. Inhibition of sperm-oocyte binding and sperm-oocyte fusion make fertilin an attractive target for the development of an immunocontraceptive vaccine. Full-length cDNAs encoding alpha and beta fertilin subunits were isolated from a rat testis cDNA library and sequenced. Using reverse transcription-polymerase chain reaction (RT-PCR), the developmental expression of fertilin alpha and beta was determined in pre-pubertal and mature rat testes. Fertilin alpha mRNA was present at all stages of development, suggesting that it is not exclusively expressed in post-meiotic germ cells. In contrast, fertilin beta mRNA was first identified in day 19 testes, coincident with the presence of pachytene spermatocytes. Polyclonal antisera raised against a 28-residue peptide (corresponding to part of the disintegrin domain) and two recombinant fusion proteins identified a 90 kDa protein in testicular sperm extracts and a 60 kDa protein in caput and cauda epididymidal sperm extracts, the predicted sizes for rat fertilin beta precursor and mature protein respectively. Indirect immunofluorescence using the anti-peptide antisera stained the acrosomal cap of permeabilized testicular, caput and caudal spermatozoa and elongating spermatids in testicular sections.


An investigation of weak syllable processing in deaf children with cochlear implants.
In this study the influence of prosodic foot structure on the processing of weak syllables in children with cochlear implants (CI) was investigated. A battery of tests investigating processing of weak syllables in single and multi-word utterances was carried out on four groups of children: 15 children with CI developing spoken language as expected (Main CI); five children with CI reported to have additional speech and language problems; 15 age matched; and 15 language matched (LM) children with normal hearing (NH). Children in the main CI and, to a lesser extent, the LM groups processed footed weak syllables preferably over unfooted weak syllables (particularly as memory load increased). Thus, these children with CI appear to possess a similar Prosodic Hierarchy (PH) to their LM peers with NH, and possibly due to the impact of delayed and constrained exposure to audition on the development of linguistic processing and short-term memory, are influenced by its foot structure in the processing of weak syllables.


A systematic review of informal caregivers' needs in providing home-based end-of-life care to people with cancer.
This paper presents the results of a systematic review examining the practical information needs of informal caregivers providing home-based palliative and end-of-life care to people with advanced cancer. Modern hospice care has led to increases in home-based palliative care services, with informal caregivers assuming responsibility for the majority of care. In response, health policy emphasises the provision of palliative care services in which both the patient and carer receive adequate support throughout illness and death. While the emotional needs of carers have been extensively researched, their practical needs with respect to the provision of physical care are yet to receive systematic attention. Systematic review. Eligible articles were identified via electronic searches of research and evidence-based databases, hand-searching of academic journals and searches of non-academic grey literature websites. Quality of research was assessed via accepted guidelines for reviewing non-randomised, observational and qualitative literature. Data were synthesised by comparing and contrasting the findings to identify prominent themes. Research consistently highlights this lack of practical support, often related to inadequate information exchange. These deficits typically manifest in relatives adopting a 'trial and error' approach to palliative care. Informal carers request a greater quantity of practically-focussed information, improvements in quality and increased methods of dissemination. Synthesis of the literature suggests that home-based palliative care services have been insufficiently focussed on assisting informal caregivers acquire practical nursing skills. Enhanced access to professional advice represents a potentially effective method of increasing carers' confidence in their ability to undertake practical aspects of home-based care. Evidence suggests that nurses and other health providers may better assist home-based carers by providing the information and skills-training necessary to facilitate this. This may necessitate the involvement of carers in the design and testing of new educational interventions.


Small sample mediation testing: misplaced confidence in bootstrapped confidence intervals.
Bootstrapping is an analytical tool commonly used in psychology to test the statistical significance of the indirect effect in mediation models. Bootstrapping proponents have particularly advocated for its use for samples of 20-80 cases. This advocacy has been heeded, especially in the Journal of Applied Psychology, as researchers are increasingly utilizing bootstrapping to test mediation with samples in this range. We discuss reasons to be concerned with this escalation, and in a simulation study focused specifically on this range of sample sizes, we demonstrate not only that bootstrapping has insufficient statistical power to provide a rigorous hypothesis test in most conditions but also that bootstrapping has a tendency to exhibit an inflated Type I error rate. We then extend our simulations to investigate an alternative empirical resampling method as well as a Bayesian approach and demonstrate that they exhibit comparable statistical power to bootstrapping in small samples without the associated inflated Type I error. Implications for researchers testing mediation hypotheses in small samples are presented. For researchers wishing to use these methods in their own research, we have provided R syntax in the online supplemental materials.


[Ultrasonographic finding and treatment of a cow with pyelonephrosis].
Ultrasonography of a six-year-old Simmental cow revealed an abscess like structure, approximately 15 cm in diameter, in place of the right kidney. The cow had a history of colic for 4 days and was referred to our clinic with a tentative diagnosis of caecal dilatation. The cow voided dark opaque urine with white floccules. Laboratory examination yielded increased haematocrit, leukocytosis with left shift, hyperbilirubinaemia and azotaemia. The diagnosis was confirmed, the caecum emptied surgically and the pus-filled structure at the site of the right kidney removed. A diagnosis of pyonephrosis was made after pathological examination of the pus-filled structure.


Interpretation of time constant and electrotonic length estimates in multicylinder or branched neuronal structures.
1. We have investigated the theoretical and practical problems associated with the interpretation of time constants and the estimation of electrotonic length with equivalent cylinder formulas for neurons best represented as multiple cylinders or branched structures. Two analytic methods were used to compute the time constants and coefficients of passive voltage transients (and time constants of current transients under voltage clamp). One method, suitable for simple geometries, involves analytic solutions to boundary value problems. The other, suitable for neurons of any geometric complexity, is an algebraic approach based on compartmental models. Neither of these methods requires the simulation of transients. 2. We computed the time constants and coefficients of voltage transients for several hypothetical neurons and also for a spinal motoneuron whose morphology was characterized from serial reconstructions. These time constants and coefficients were used to generate voltage transients. Then exponential peeling, nonlinear regression, and transform methods were applied to these transients to test how well these procedures estimate the underlying time constants and coefficients. 3. For a serially reconstructed motoneuron with 732 compartments, we found that the theoretical and peeled tau 0 values were nearly equal, but the theoretical tau 1 was much larger than the peeled tau 1. The theoretical tau 1 could not be peeled because it was associated with a coefficient, C1, that had a very small value. In fact, there were 156 time constants between 1.0 and 6.0 ms, most of which had very small coefficients; none had a coefficient larger than 2% of the signal. The peeled value of tau 1 (called tau 1 peel) can be viewed as some sort of a weighted average of the time constants having the largest coefficients. 4. We studied simple hypothetical neurons to determine what interpretation could be applied to the multitude of theoretical time constants. We found that after tau 0, there was a group of time constants associated with eigenfunctions that were odd (or approximately odd) functions with respect to the soma. These time constants could be interpreted as "equalizing" time constants along particular paths between different pairs of dendritic terminals in the neuron. After this group of time constants, there was one that we call tau even because it was associated with an eigen-function that was approximately even with respect to the soma. This tau even could be interpreted as an equalizing time constant for charge equalization between proximal membrane (soma and proximal dendrites) and distal membrane (including all distal dendrites).4=


Child abuse: approach and management.
Child abuse is a common diagnosis in the United States and should be considered any time neglect or emotional, physical, or sexual abuse is a possibility. Although home visitation programs have been effective in preventing child maltreatment, much of the approach to and management of child abuse is directed by expert opinion or legal mandate. Any suspicion of abuse must be reported to Child Protective Services. A multidisciplinary approach is recommended to adequately evaluate and treat child abuse victims; however, the responsibility often lies with the family physician to recognize and treat these cases at first presentation to prevent significant morbidity and mortality.


Two minor determinants of myelin basic protein induce experimental allergic encephalomyelitis in SJL/J mice.
Experimental allergic encephalomyelitis (EAE) is an autoimmune demyelinating disease of the central nervous system (CNS) that occurs after immunization of animals with myelin basic protein (MBP). The disease is a prototype model for the study of antigen-specific T helper cell-mediated autoimmune disease. In SJL/J mice, EAE is mediated by T helper cells directed against a 40-amino acid COOH-terminal peptic fragment of mouse small MBP. To identify the minimal T cell epitopes of MBP responsible for EAE, overlapping peptides completely encompassing the epitopes within this region were synthesized. A 28-residue peptide of mouse MBP spanning residues 87-114 (pM87-114) was able to elicit both a strong T cell response and chronic relapsing disease. To better localize the T cell epitopes, shorter peptides within this region were synthesized and two overlapping peptides, pM87-98 and pM91-104, were able to induce EAE. T cell clones and bulk lymph node cell populations reactive with pM87-98 did not respond to pM91-104. However, lymph node cells reactive with pM91-104 also reacted with pM87-98, thus showing that these two peptides represent contiguous, but distinct encephalitogenic epitopes and that both these epitopes may be contained within pM87-98. In addition, pM87-114 and pM87-98 were found to be minor determinants of the total T cell response to rat and rabbit MBP. The restricted response to MBP in SJL/J mice is similar to that of the PL/J mice in that each appears to have only a single peptide region in MBP that elicits encephalitogenic T cells. However, within the region studied, there were two if not more T cell epitopes. This differs from the single encephalitogenic PL/J epitope. These findings of a single encephalitogenic peptide region with multiple T cell epitopes and the fact that encephalitogenic T cell epitopes may be subdominant have implications for the design of treatments directed at the T cell receptor-MHC-peptide epitope complex in autoimmune disease.


Tolerance of T cell receptor gamma/delta cells in the intestine.
The present study examined the mechanism(s) of tolerance induction for intestinal intraepithelial lymphocytes (iIELs) using an alloantigen (Ag)-specific gamma/delta T cell receptor (TCR gamma/delta) transgenic (Tg) model. In Tg Ag-bearing H-2b/d mice (Tgb/d), Tg iIELs were Thy-1-, CD44+, CD45R (B220)+, and CD5+, whereas in syngeneic Tgd/d mice, iIELs were Thy-1+, CD44-, and CD45R- with a subset of CD5+ cells. Previously, we had shown that tolerance for Tgb/d iIELs involved functional anergy and deletion (Barrett, T. A., M. L. Delvy, D. M. Kennedy, L. Lefrancois, L. A. Matis, A. L. Dent, S. M. Hedrick, and J. A. Bluestone. 1992. J. Exp. Med. 175:65). In this study we demonstrate that Tgb/d iIELs expressing dull levels of Thy-1 proliferated in the presence of exogenous rIL-2. A direct precursor-product relationship between the Thy-1+-responsive iIELs and the tolerant Thy-1dul/- iIELs was demonstrated by adoptive transfer into severe combined immunodeficient (SCID) mice. Tg Thy-1+ iIELs reconstituting Ag+ but not Ag- SCID mice downregulated Thy-1 after Ag exposure in vivo. Analysis of bone marrow (BM) chimeras demonstrated the persistence of Tg IELs in all Ag+ chimeras although a modest degree of clonal deletion was apparent. The greatest percentage of Tg IELs were detected when Ag was restricted to radioresistant cells (e.g., epithelial cells) compared with BM-derived antigen-presenting cells (APC). This was especially apparent in thymectomized chimeric mice. Consistent with the notion that Ag-bearing epithelial cells may be poor APC, isolated intestinal epithelial cells from Ag-bearing mice failed to stimulate Tg iIELs compared with splenic APC. These studies suggest that the major population of TCR gamma/delta iIELs were probably extrathymically derived and encountered self-Ag on intestinal epithelial cells. The induction of tolerance likely involved an activation event resulting in downregulation of Thy-1. These mechanisms of tolerance for TCR gamma/delta iIELs led to the persistence of a reservoir of self-reactive T cells with the potential for mediating autoimmune disease.


Tracking iron-associated proteomes in pathogens by a fluorescence approach.
Porphyromonas gingivalis is a key oral anaerobic bacterium involved in human periodontitis, which may affect up to 15% adults worldwide. Using the membrane permeable fluorescent probe Fe-TRACER, we identified 17 iron-associated proteins in Porphyromonas gingivalis. We demonstrated the specific binding of the probe towards iron-associated proteins using transferrin as an example and provided an X-ray structure of the fluorescent probe-bound transferrin. Our study provides a basis for the understanding of iron homeostasis in pathogens, and our approach based on the integration of fluorescence imaging with proteomics and bioinformatics can be readily extended to mine other metalloproteomes in microbials.


MAPRE1 as a plasma biomarker for early-stage colorectal cancer and adenomas.
Blood-based biomarkers for early detection of colorectal cancer could complement current approaches to colorectal cancer screening. We previously identified the APC-binding protein MAPRE1 as a potential colorectal cancer biomarker. Here, we undertook a case-control validation study to determine the performance of MAPRE1 in detecting early colorectal cancer and colon adenoma and to assess the potential relevance of additional biomarker candidates. We analyzed plasma samples from 60 patients with adenomas, 30 with early colorectal cancer, 30 with advanced colorectal cancer, and 60 healthy controls. MAPRE1 and a set of 21 proteins with potential biomarker utility were assayed using high-density antibody arrays, and carcinoembryonic antigen (CEA) was assayed using ELISA. The biologic significance of the candidate biomarkers was also assessed in colorectal cancer mouse models. Plasma MAPRE1 levels were significantly elevated in both patients with adenomas and patients with colorectal cancer compared with controls (P < 0.0001). MAPRE1 and CEA together yielded an area under the curve of 0.793 and a sensitivity of 0.400 at 95% specificity for differentiating early colorectal cancer from controls. Three other biomarkers (AK1, CLIC1, and SOD1) were significantly increased in both adenoma and early colorectal cancer patient plasma samples and in plasma from colorectal cancer mouse models at preclinical stages compared with controls. The combination of MAPRE1, CEA, and AK1 yielded sensitivities of 0.483 and 0.533 at 90% specificity and sensitivities of 0.350 and 0.467 at 95% specificity for differentiating adenoma and early colorectal cancer, respectively, from healthy controls. These findings suggest that MAPRE1 can contribute to the detection of early-stage colorectal cancer and adenomas together with other biomarkers.


Reduced dendritic arborization and hyperexcitability of pyramidal neurons in a Scn1b-based model of Dravet syndrome.
Epileptic encephalopathies, including Dravet syndrome, are severe treatment-resistant epilepsies with developmental regression. We examined a mouse model based on a human β1 sodium channel subunit (Scn1b) mutation. Homozygous mutant mice shared phenotypic features and pharmaco-sensitivity with Dravet syndrome. Patch-clamp analysis showed that mutant subicular and layer 2/3 pyramidal neurons had increased action potential firing rates, presumably as a consequence of their increased input resistance. These changes were not seen in L5 or CA1 pyramidal neurons. This raised the concept of a regional seizure mechanism that was supported by data showing increased spontaneous synaptic activity in the subiculum but not CA1. Importantly, no changes in firing or synaptic properties of gamma-aminobutyric acidergic interneurons from mutant mice were observed, which is in contrast with Scn1a-based models of Dravet syndrome. Morphological analysis of subicular pyramidal neurons revealed reduced dendritic arborization. The antiepileptic drug retigabine, a K+ channel opener that reduces input resistance, dampened action potential firing and protected mutant mice from thermal seizures. These results suggest a novel mechanism of disease genesis in genetic epilepsy and demonstrate an effective mechanism-based treatment of the disease.


A Novel Video-Based Patient Education Program to Reduce Penile Prosthetic Surgery Cancellations.
Penile prosthetic surgery is an effective treatment for men with erectile dysfunction. Cancellation of surgery is disruptive and costly to patients, physicians, and the healthcare system. This pilot study sought to analyze surgery cancellations and implement a video-based patient education program to decrease surgery noncompletion. Baseline penile prosthetic surgery completion, rescheduling, and cancellation rates among consecutively scheduled surgeries were determined using a national cohort. Selected prosthetic surgeons then implemented Vidscrip, a video-based patient education program. Prerecorded videos were delivered via text message 14 days, 7 days, and 1 day preoperatively, as well as 1 day postoperatively. Subsequent analysis determined noncompletion rates, reasons for noncompletion, surgeon volume, and video utilization. Two-hundred twenty-six surgeries were scheduled in the baseline cohort; 141 were completed, and 85 were rescheduled or canceled. Among the intervention cohort, 290 patients completed, 7 rescheduled, and 37 canceled surgery. After program implementation, the surgery noncompletion rate was reduced compared to baseline (13.2% vs. 37.6%, p < .05), corresponding to a number needed to treat of 4.1. When stratified by surgeon volume, there was no difference in noncompletion rate (>20 cases vs. ≤20 cases: 8.20% vs. 32.0%, p = .35). Video utilization was widely variable among practices (median viewing time 58.6 min, IQR 5.09-113). Penile prosthetic surgery is frequently rescheduled or canceled. Implementing a video-based patient education program reduces surgery noncompletion, improving efficiency and quality of care. Wider implementation is needed to validate these findings, while cost-effectiveness analyses may further support their broad adoption.


Management of Severe Bleeding in Patients Treated with Direct Oral Anticoagulants: An Observational Registry Analysis.
The use of prothrombin complex concentrates and the role of plasma concentration of anticoagulants in the management of bleeding in patients treated with direct oral anticoagulants are still debated. Our aim was to describe management strategies and outcomes of severe bleeding events in patients treated with direct oral anticoagulants. We performed a prospective cohort study of 732 patients treated with dabigatran, rivaroxaban, or apixaban hospitalized for severe bleeding, included prospectively in the registry from June 2013 to November 2015. Bleeding was gastrointestinal or intracranial in 37% (212 of 732) and 24% (141 of 732) of the cases, respectively. Creatinine clearance was lower than 60 ml/min in 61% (449 of 732) of the cases. The plasma concentration of direct oral anticoagulants was determined in 62% (452 of 732) of the cases and was lower than 50 ng/ml or higher than 400 ng/ml in 9.2% (41 of 452) and in 6.6% (30 of 452) of the cases, respectively. Activated or nonactivated prothrombin complex concentrates were administered in 38% of the cases (281 of 732). Mortality by day 30 was 14% (95% CI, 11 to 16). Management of severe bleeding in patients treated with direct oral anticoagulants appears to be complex. The use of prothrombin complex concentrates differs depending on bleeding sites and direct oral anticoagulant plasma concentrations. Mortality differs according to bleeding sites and was similar to previous estimates.


Conservation in a cup of water: estimating biodiversity and population abundance from environmental DNA.
Three mantras often guide species and ecosystem management: (i) for preventing invasions by harmful species, 'early detection and rapid response'; (ii) for conserving imperilled native species, 'protection of biodiversity hotspots'; and (iii) for assessing biosecurity risk, 'an ounce of prevention equals a pound of cure.' However, these and other management goals are elusive when traditional sampling tools (e.g. netting, traps, electrofishing, visual surveys) have poor detection limits, are too slow or are not feasible. One visionary solution is to use an organism's DNA in the environment (eDNA), rather than the organism itself, as the target of detection. In this issue of Molecular Ecology, Thomsen et al. (2012) provide new evidence demonstrating the feasibility of this approach, showing that eDNA is an accurate indicator of the presence of an impressively diverse set of six aquatic or amphibious taxa including invertebrates, amphibians, a fish and a mammal in a wide range of freshwater habitats. They are also the first to demonstrate that the abundance of eDNA, as measured by qPCR, correlates positively with population abundance estimated with traditional tools. Finally, Thomsen et al. (2012) demonstrate that next-generation sequencing of eDNA can quantify species richness. Overall, Thomsen et al. (2012) provide a revolutionary roadmap for using eDNA for detection of species, estimates of relative abundance and quantification of biodiversity.


Evidence that the 16 kDa proteolipid (subunit c) of the vacuolar H(+)-ATPase and ductin from gap junctions are the same polypeptide in Drosophila and Manduca: molecular cloning of the Vha16k gene from Drosophila.
The 16 kDa proteolipid (subunit c) of the eukaryotic vacuolar H(+)-ATPase (V-ATPase) is closely related to the ductin polypeptide that forms the connexon channel of gap junctions in the crustacean Nephrops norvegicus. Here we show that the major protein component of Manduca sexta gap junction preparations is a 16 kDa polypeptide whose N-terminal sequence is homologous to ductin and is identical to the deduced sequence of a previously cloned cDNA from Manduca (Dow et al., Gene, 122, 355-360, 1992). We also show that a Drosophila melanogaster cDNA, highly homologous to the Manduca cDNA, can rescue Saccharomyces cerevisiae, defective in V-ATPase function, in which the corresponding yeast gene, VMA3, has been inactivated. Evidence is presented for a single genetic locus (Vha16) in Drosophila, which in adults at least contains a single transcriptional unit. Taken together, the data suggest that in Drosophila and Manduca, the same polypeptide is both the proteolipid subunit c component of the V-ATPase and the ductin component of gap junctions. The intron/exon structure of the Drosophila Vha16 is identical to that of a human Vha16 gene, and is consistent with an ancient duplication of an 8 kDa domain. A pilot study for gene inactivation shows that transposable P-elements can be easily inserted into the Drosophila ductin Vha16 gene. Although without phenotypic consequences, these can serve as a starting point for generation of null alleles.


Paludibaculum fermentans gen. nov., sp. nov., a facultative anaerobe capable of dissimilatory iron reduction from subdivision 3 of the Acidobacteria.
A facultatively anaerobic, non-pigmented, non-spore-forming bacterium was isolated from a littoral wetland of a boreal lake located on Valaam Island, northern Russia, and designated strain P105(T). Cells of this isolate were Gram-negative, non-motile rods coated by S-layers with p2 lattice symmetry. Sugars were the preferred growth substrates. Under anoxic conditions, strain P105(T) was capable of fermentation and dissimilatory Fe(III) reduction. End products of fermentation were acetate, propionate and H2. Strain P105(T) was a mildly acidophilic, mesophilic organism, capable of growth at pH 4.0-7.2 (optimum pH 5.5-6.0) and at 4-35 °C (optimum at 20-28 °C). The major fatty acids were iso-C(15 : 0) and C(16 : 1)ω7c; the cells also contained significant amounts of 13,16-dimethyl octacosanedioic acid (isodiabolic acid). The major polar lipids were phosphocholine and phosphoethanolamine; the quinone was MK-8. The G+C content of the DNA was 60.5 mol%. 16S rRNA gene sequence analysis showed that strain P105(T) belongs to subdivision 3 of the Acidobacteria and is only distantly related (90% sequence similarity) to the only currently characterized member of this subdivision, Bryobacter aggregatus. The novel isolate differs from Bryobacter aggregatus in its cell morphology and ability to grow under anoxic conditions and in the presence of iron- and nitrate-reducing capabilities as well as quinone and polar lipid compositions. These differences suggest that strain P105(T) represents a novel genus and species, for which the name Paludibaculum fermentans gen. nov., sp. nov., is proposed. The type strain of Paludibaculum fermentans is P105(T) ( = DSM 26340(T) = VKM B-2878(T)).


Hyperinsulinemia, risk factors, and coronary heart disease. The Zutphen Elderly Study.
We investigated the association between fasting insulin concentration--an indicator of insulin resistance in nondiabetic individuals--cardiovascular risk factors, and coronary heart disease in a study of 390 men in the town of Zutphen. In 1990, an extensive examination was carried out on the participating men (aged 70 to 89 years). Fasting insulin levels were determined and a number of other risk factors measured. Known and newly diagnosed diabetics were excluded from the data analyses. Fasting insulin concentration was significantly associated with levels of glucose, triglycerides, uric acid, serum albumin, creatinine, and fibrinogen as well as resting heart rate. Inverse associations with high-density lipoprotein cholesterol and factor VII activity were observed. These results were independent of confounding factors such as age, body mass index, ratio of subscapular to triceps skinfold thicknesses, cigarette smoking, physical activity, and alcohol consumption. Men with a fasting insulin level higher than 80 pmol/L (highest quartile of the distribution) had a significantly higher prevalence of coronary heart disease and especially of myocardial infarction. This result was independent of potential confounding variables as well as of possible intermediates (total and high-density lipoprotein cholesterol, hypertension, serum triglycerides, fasting glucose, and other risk factors related to fasting insulin) (odds ratio, 2.2; 95% confidence interval, 1.2-4.0). No association between fasting insulin level and hypertension or blood pressure was observed. These results show that fasting insulin is an important indicator of coronary heart disease in elderly men. Clotting factors, resting heart rate, uric acid, serum albumin, and creatinine may also play a role in this metabolic syndrome.


Mental performance in 8-year-old children fed reduced protein content formula during the 1st year of life: safety analysis of a randomised clinical trial.
In humans, maximum brain development occurs between the third trimester of gestation and 2 years of life. Nutrition during these critical windows of rapid brain development might be essential for later cognitive functioning and behaviour. In the last few years, trends on protein recommendations during infancy and childhood have tended to be lower than that in the past. It remains to be demonstrated that lower protein intakes among healthy infants, a part of being able to reduce obesity risk, is safe in terms of mental performance achievement. Secondary analyses of the EU CHOP, a clinical trial in which infants from five European countries were randomised to be fed a higher or a lower protein content formula during the 1st year of life. Children were assessed at the age of 8 years with a neuropsychological battery of tests that included assessments of memory (visual and verbal), attention (visual, selective, focused and sustained), visual-perceptual integration, processing speed, visual-motor coordination, verbal fluency and comprehension, impulsivity/inhibition, flexibility/shifting, working memory, reasoning, visual-spatial skills and decision making. Internalising, externalising and total behaviour problems were assessed using the Child Behaviour Checklist 4-18. Adjusted analyses considering factors that could influence neurodevelopment, such as parental education level, maternal smoking, child's gestational age at birth and head circumference, showed no differences between feeding groups in any of the assessed neuropsychological domains and behaviour. In summary, herewith we report on the safety of lower protein content in infant formulae (closer to the content of human milk) according to long-term mental performance.


The performance of an automatic acoustic-based program classifier compared to hearing aid users' manual selection of listening programs.
To compare preference for and performance of manually selected programmes to an automatic sound classifier, the Phonak AutoSense OS. A single blind repeated measures study. Participants were fit with Phonak Virto V90 ITE aids; preferences for different listening programmes were compared across four different sound scenarios (speech in: quiet, noise, loud noise and a car). Following a 4-week trial preferences were reassessed and the users preferred programme was compared to the automatic classifier for sound quality and hearing in noise (HINT test) using a 12 loudspeaker array. Twenty-five participants with symmetrical moderate-severe sensorineural hearing loss. Participant preferences of manual programme for scenarios varied considerably between and within sessions. A HINT Speech Reception Threshold (SRT) advantage was observed for the automatic classifier over participant's manual selection for speech in quiet, loud noise and car noise. Sound quality ratings were similar for both manual and automatic selections. The use of a sound classifier is a viable alternative to manual programme selection.


Association between early and late response in eyes with central or hemiretinal vein occlusion treated with anti-VEGF agents : SCORE2 report 12: secondary analysis of the SCORE2 clinical trial.
To assess whether early visual acuity letter score change from baseline (ΔVALS) and early spectral domain optical coherence tomography (SD-OCT) measures of center point thickness (CPT) are associated with later ΔVALS in eyes with macular edema due to central or hemiretinal vein occlusion treated with intravitreal aflibercept or bevacizumab. Secondary analysis of a randomized clinical trial of 362 participants. Considered separately at month 3, CPT (categorized as ≤ 300 μm, > 300 μm) and ΔVALS (categorized as < 5, 5-9, ≥ 10) are predictive of ΔVALS at month 6 (aflibercept: P = 0.02 for CPT and P < 0.0001 for ΔVALS; bevacizumab: P = 0.007 for CPT and P < 0.0001 for ΔVALS) and, except for CPT in the bevacizumab arm, also predictive of ΔVALS at month 12 (aflibercept: P = 0.03 for CPT and P < 0.0001 for ΔVALS; bevacizumab: P = 0.18 for CPT and P < 0.0001 for ΔVALS). Month 3 predictors are also associated with average ΔVALS from months 4 to 12 (CPT P = 0.01 in the aflibercept arm, P = 0.02 in the bevacizumab arm; ΔVALS > 10 versus < 5; P < 0.001 for both aflibercept and bevacizumab). When month 3 measures are considered jointly, ΔVALS effect remains significant for average ΔVALS from months 4 to 12 (aflibercept: P = 0.002; bevacizumab: P < 0.0001) but not CPT (aflibercept: P = 0.18; bevacizumab: P = 0.22). While both month 3 ΔVALS and CPT are predictive of ΔVALS after month 3 through month 12, early ΔVALS has a stronger relationship than CPT with later ΔVALS. SCORE2 registration number is NCT01969708.


A method for assessing the damping characteristics of periodontal tissues: goals and limitations.
The Periotest method, an objective, noninvasive clinical diagnostic method, is a dynamic procedure that measures the resistance of the periodontium to a defined impact load. It has been reported that Periotest values depend to some extent on tooth mobility, but mainly on the damping characteristics of the periodontium. Nevertheless, the real clinical meaning of the measurements and some important limitations of the Periotest measuring principle still seem to be poorly understood. In the present study, the relationship between damping characteristics of periodontal tissues and tooth mobility was investigated. The best correlations between tooth deflection and Periotest values were found for teeth showing a certain degree of clinical mobility (R2 from .79 to .91). Nevertheless, this correlation was clearly lower when only healthy subjects were examined (R2 from .43 to .54). The better correlation found for forces greater than 1.0 N indicates that the damping characteristics assessed with the Periotest method are related to secondary tooth movement. The Periotest methodology, measuring principle, and limitations are critically reviewed.


Nanostructural origin of blue fluorescence in the mineral karpatite.
The colour of crystals is a function of their atomic structure. In the case of organic crystals, it is the spatial relationships between molecules that determine the colour, so the same molecules in the same arrangement should produce crystals of the same colour, regardless of whether they arise geologically or synthetically. There is a naturally-occurring organic crystal known as karpatite which is prized for its beautiful blue fluorescence under ultra-violet illumination. When grown under laboratory conditions however, the crystals fluoresce with an intense green colour. For 20 years, this difference has been thought to be due to chemical impurities in the laboratory-grown material. Using electron microscopy coupled with fluorescence spectroscopy and X-Ray diffraction, we report here that this disparity is instead due to differences in the structure of the crystals at the nanoscale. The results show that in nature, karpatite has a nanotexture that is not present in the synthetic crystals, which enables different photonic pathways and therefore a blue, rather than green colour whilst undergoing fluorescence.


Vitellogenin underwent subfunctionalization to acquire caste and behavioral specific expression in the harvester ant Pogonomyrmex barbatus.
The reproductive ground plan hypothesis (RGPH) proposes that the physiological pathways regulating reproduction were co-opted to regulate worker division of labor. Support for this hypothesis in honeybees is provided by studies demonstrating that the reproductive potential of workers, assessed by the levels of vitellogenin (Vg), is linked to task performance. Interestingly, contrary to honeybees that have a single Vg ortholog and potentially fertile nurses, the genome of the harvester ant Pogonomyrmex barbatus harbors two Vg genes (Pb_Vg1 and Pb_Vg2) and nurses produce infertile trophic eggs. P. barbatus, thus, provides a unique model to investigate whether Vg duplication in ants was followed by subfunctionalization to acquire reproductive and non-reproductive functions and whether Vg reproductive function was co-opted to regulate behavior in sterile workers. To investigate these questions, we compared the expression patterns of P. barbatus Vg genes and analyzed the phylogenetic relationships and molecular evolution of Vg genes in ants. qRT-PCRs revealed that Pb_Vg1 is more highly expressed in queens compared to workers and in nurses compared to foragers. By contrast, the level of expression of Pb_Vg2 was higher in foragers than in nurses and queens. Phylogenetic analyses show that a first duplication of the ancestral Vg gene occurred after the divergence between the poneroid and formicoid clades and subsequent duplications occurred in the lineages leading to Solenopsis invicta, Linepithema humile and Acromyrmex echinatior. The initial duplication resulted in two Vg gene subfamilies preferentially expressed in queens and nurses (subfamily A) or in foraging workers (subfamily B). Finally, molecular evolution analyses show that the subfamily A experienced positive selection, while the subfamily B showed overall relaxation of purifying selection. Our results suggest that in P. barbatus the Vg gene underwent subfunctionalization after duplication to acquire caste- and behavior- specific expression associated with reproductive and non-reproductive functions, supporting the validity of the RGPH in ants.


The relationship between obesity and fecundity.
Obesity is an important factor that might reduce fecundity. In order to determine the underlying physiological mechanisms and risk factors, the obesity-fecundity association is investigated in relation to parity, menstrual cycle regularity, smoking habits, and age. This was a retrospective cohort study of 22,840 women who gave birth between January 2006 and January 2007 in the Dr Zekai Tahir Burak Women's Health Research and Education Hospital. Age, parity, prepregnancy body mass index (BMI) values, time to pregnancy data related to smoking, and reproductive, medical, and gynecological history were obtained from the medical records. Fecundity was reduced for overweight and obese women compared with optimal weight women, and this reduction was more evident for obese primiparous women. Fecundity remained reduced for overweight and obese women with normal menstrual cycles. Obese and overweight women were found to smoke significantly more than the optimal weight group. Obesity was found to be associated with reduced fecundity for all weight-adjusted groups of women and persisted for women with regular cycles. Weight loss should be encouraged initially during the treatment of infertile overweight and obese women.


A foundation for provitamin A biofortification of maize: genome-wide association and genomic prediction models of carotenoid levels.
Efforts are underway for development of crops with improved levels of provitamin A carotenoids to help combat dietary vitamin A deficiency. As a global staple crop with considerable variation in kernel carotenoid composition, maize (Zea mays L.) could have a widespread impact. We performed a genome-wide association study (GWAS) of quantified seed carotenoids across a panel of maize inbreds ranging from light yellow to dark orange in grain color to identify some of the key genes controlling maize grain carotenoid composition. Significant associations at the genome-wide level were detected within the coding regions of zep1 and lut1, carotenoid biosynthetic genes not previously shown to impact grain carotenoid composition in association studies, as well as within previously associated lcyE and crtRB1 genes. We leveraged existing biochemical and genomic information to identify 58 a priori candidate genes relevant to the biosynthesis and retention of carotenoids in maize to test in a pathway-level analysis. This revealed dxs2 and lut5, genes not previously associated with kernel carotenoids. In genomic prediction models, use of markers that targeted a small set of quantitative trait loci associated with carotenoid levels in prior linkage studies were as effective as genome-wide markers for predicting carotenoid traits. Based on GWAS, pathway-level analysis, and genomic prediction studies, we outline a flexible strategy involving use of a small number of genes that can be selected for rapid conversion of elite white grain germplasm, with minimal amounts of carotenoids, to orange grain versions containing high levels of provitamin A.


Identification of the molecular defect in a family with spondyloepiphyseal dysplasia.
Spondyloepiphyseal dysplasias (SED) are a heterogeneous group of inherited disorders characterized by disproportionate short stature and pleiotropic involvement of the skeletal and ocular systems. Evidence has suggested that SED may result from structural defects in type II collagen. To confirm the validity of this hypothesis, the structure of the "candidate" type II collagen gene (COL2A1) has been directly examined in a relatively large SED family. Coarse scanning of the gene by Southern blot hybridization identified an abnormal restriction pattern in one of the affected members of the kindred. Analysis of selected genomic fragments, amplified by the polymerase chain reaction, precisely localized the molecular defect and demonstrated that all affected family members carried the same heterozygous single-exon deletion. As a consequence of the mutation, nearly 90 percent of the assembled type II collagen homotrimers are expected to contain one or more procollagen subunits harboring an interstitial deletion of 36 amino acids in the triple helical domain.


Inhibition of chronic prostate inflammation by hyaluronic acid through an immortalized human prostate stromal cell line model.
Benign prostatic hyperplasia (BPH) is the most common urologic disease among elderly men. A well-established in vitro cell model is required to determine the therapeutic mechanism of BPH inflammation. In this study, we attempted to establish an immortalized human prostate stromal cell line by transfecting with HPV-16 E6/E7 and designated as ihPSC. No significant difference was found in fibroblast-like morphology between primary hPSC and ihPSC. The ihPSC possessed a significantly higher cell proliferation rate than primary hPSC. The prostate-specific markers and proteins including cytoskeleton (α-SMA and vimentin) and smooth muscle (calponin), especially the androgen receptor (AR) were also examined in ihPSC, almost identical to the primary hPSC. To create an in vitro model featuring chronic prostatic inflammation, ihPSC was stimulated with IFN-γ+IL-17 and then treated with the high molecular weight hyaluronic acid hylan G-F 20 as an alternative strategy for inhibiting BPH inflammation. Hylan G-F 20 could dose-dependently diminish the inflammation-induced proliferation in ihPSC. The enhanced expressions of inflammatory molecules including IL-1β, IL-6, IL-8, cyclooxygenase 2 (COX2), inducible nitrogen oxide synthase (iNOS), and Toll-like receptor 4 (TLR4) were all abolished by hylan G-F 20. For inflammatory signaling, hylan G-F 20 can also diminish the IFN-γ+IL-17-increased expression of iNOS and p65 in ihPSC. These findings suggest that ihPSC could provide a mechanism-based platform for investigating prostate inflammation. The hylan G-F 20 showed strong anti-inflammatory effects by decreasing inflammatory cytokines and signalings in the ihPSC, indicating its therapeutic potentials in BPH treatment in the future.


Growing hierarchical tree SOM: an unsupervised neural network with dynamic topology.
In this paper we introduce a tree structured self-organizing network, called the Growing Hierarchical Tree SOM (GHTSOM), that combines unsupervised learning with a dynamic topology for hierarchical classification of unlabelled data sets. The main feature of the proposed model is a SOM-like self-organizing process that allows the network to adapt the topology of each layer of the hierarchy to the characteristics of the training set. In particular the self-organization is obtained in two steps: the first one concerns the learning phase and is finalized with the creation of a tree of SOMs, while the second one is in regard to the clustering phase and provides the formation of classes for each level of the tree (hence self-organization not only for training but also for the creation of topological connections). As a result the network works without the need for user-defined parameters. Experimental results are proposed on both synthetic and real data sets.


Comorbidity of chronic diseases: effects of disease pairs on physical and mental functioning.
Although much research has been done on the impact of chronic illness on quality of life, still relatively little is known about the role of comorbidity. Given the growing number of (older) people with multiple chronic conditions, more information is needed on the effects of specific disease combinations for preventive purposes. In a nationwide representative sample of 1673 non-institutionalized chronic disease patients (recruited in 56 general practices) in The Netherlands, we assessed the separate and joint effects of cardiovascular disease, cancer, arthritis, chronic respiratory disease, diabetes mellitus, and thyroid dysfunction on physical and mental functioning. Data on medical diagnoses were provided by the general practitioners; data on physical and mental functioning were collected by a patient survey (SF-36). Compared to reference data of the general population, physical functioning appeared to be worse in all six diagnostic groups, whereas mental functioning was more or less comparable. Patients with arthritis or those suffering from comorbidity reported the lowest levels of physical functioning. Synergistic effects of combinations of diabetes, cardiovascular disease and/or chronic respiratory disease were found, indicating that patients suffering from these disease combinations run a higher risk of physical disability than could be expected from their separate effects.


Theoretical and Empirical Foundations for a Unified Pyramid of Human Motivation.
Revisions are proposed to the taxonomic model of human motivation of Forbes (Review of General Psychology, 15(2), 85-98, 2011) in order to incorporate a heretofore missing fourth life domain, the spiritual. The growing literature on spiritual motives is systematically reviewed in accordance with literature review standards for theory development (Templier & Paré, 2018) focusing on the objective of identifying comprehensive theoretical systems that explicitly incorporate the spiritual domain as one of a limited set of human life domains. The structure of the Forbes model is contrasted with thirteen theoretical systems that explicitly incorporate the spiritual as a fourth life domain. Consistent with the Forbes model, the spiritual domain is proposed to consist of three modes of existence (Being, Doing, Having) represented as justice motivation, moral motivation, and transcendental motivation, respectively, as well as both promotion and prevention goals within each of the three motives. Empirical evidence is reviewed in support of a revised heuristic device wherein the Spiritual domain is closely linked with the Intrapsychic and Interpersonal domains, but not the Instrumental domain, resulting in a pyramidal structure and corresponding set of testable hypotheses.


Unengaged vertex in nulliparous women in active labor. A risk factor for cesarean delivery.
To compare the route of delivery among nulliparous parturients with and without an engaged vertex in the early, active phase of labor. Prospectively, the position of the fetal head was ascertained among nulliparous women at 37 weeks' gestation or more in early, active labor (cervical dilation > or = 4 cm with adequate contractions). Sixteen variables, including maternal demographics, obstetric complications and intrapartum course, were examined using chi 2 and logistic regression analysis. Among the 77 patients, 33 (42.8%) had an unengaged vertex and 44 (57.2%) had an engaged vertex in active labor. Of the 22 cesarean deliveries for arrest disorder, 2 were in the engaged and 20 in the unengaged group (P < .001). The mean birth weight was similar among those who had vaginal (3,211 +/- 416 g) and cesarean delivery (3,400 +/- 489 g, P = .08). Univariate analysis indicated that chorioamnionitis (relative risk [RR] 2.6, 95% confidence interval [CI] 1.4-4.9) and unengaged vertex (RR 13.3, CI 3.3-53.0) were associated with cesarean delivery for arrest disorders. When entered into a multiple logistic model, only unengagement was a risk factor for cesarean delivery. The following were not associated with cesarean delivery: maternal demographics, gestational age, estimate of fetal weight, presence or absence of meconium, preeclampsia, diabetes mellitus, private obstetric care or use of epidural anesthesia. Among nulliparous parturients, an unengaged vertex is a significant risk factor for cesarean delivery for arrest disorders.


Dysphagia, nutrition, and hydration in ischemic stroke patients at admission and discharge from acute care.
Dysphagia may predispose stroke patients toward undernutrition and hydration. These comorbidities increase patient risks for reduced functional outcome and short-term mortality. Despite this impact, available information on relationships among dysphagia, nutrition, and hydration status in acute stroke is limited and conflicted. This study evaluated nutrition and hydration status in ischemic stroke patients with versus without clinically significant dysphagia at admission and at discharge from acute care. Sixty-seven patients admitted to the stroke unit in a tertiary-care hospital provided data for this study. On the day of hospital admission and upon discharge or at 7 days post admission, serum biochemical measures were obtained for nutrition (prealbumin) and hydration status (BUN/Cr). Clinical evaluation for dysphagia, nutrition status, and stroke severity were completed an average of 1.4 days following hospital admission. Dysphagia was identified in 37 % of the cohort. At admission 32 % of patients demonstrated malnutrition based on prealbumin levels and 53 % demonstrated evidence of dehydration based on BUN/Cr levels. No differences in nutrition status were attributed to dysphagia. Patients with dysphagia demonstrated significantly higher BUN/Cr levels (greater dehydration) than patients without dysphagia at admission and at discharge. Dehydration at both admission and discharge was associated with dysphagia, clinical nutrition status, and stroke severity. Results of this study support prior results indicating that dysphagia is not associated with poor nutrition status during the first week post stroke. Dehydration status is associated with dysphagia during this period. The results have implications for future confirmatory research and for clinical management of dysphagia in the acute stroke period.


The HXT1 gene product of Saccharomyces cerevisiae is a new member of the family of hexose transporters.
Two novel genes affecting hexose transport in the yeast Saccharomyces cerevisiae have been identified. The gene HXT1 (hexose transport), isolated from plasmid pSC7, was sequenced and found to encode a hydrophobic protein which is highly homologous to the large family of sugar transporter proteins from eucaryotes and procaryotes. Multicopy expression of the HXT1 gene restored high-affinity glucose transport to the snf3 mutant, which is deficient in a significant proportion of high-affinity glucose transport. HXT1 was unable to complement the snf3 growth defect in low copy number. The HXT1 protein was found to contain 12 putative membrane-spanning domains with a central hydrophilic domain and hydrophilic N- and C-terminal domains. The HXT1 protein is 69% identical to GAL2 and 66% identical to HXT2, and all three proteins were found to have a putative leucine zipper motif at a consensus location in membrane-spanning domain 2. Disruption of the HXT1 gene resulted in loss of a portion of high-affinity glucose and mannose transport, and wild-type levels of transport required both the HXT1 and SNF3 genes. Unexpectedly, expression of beta-galactosidase activity by using a fusion of the lacZ gene to the HXT1 promoter in a multicopy plasmid was maximal during lag and early exponential phases of growth, decreasing approximately 100-fold upon further entry into exponential growth. Deletion analysis of pSC7 revealed the presence of another gene (called ORF2) capable of suppressing the snf3 null mutant phenotype by restoring high-affinity glucose transport and increased low-affinity transport.


Assessing food security using household consumption expenditure surveys (HCES): a scoping literature review.
To meet some of the UN's seventeen Sustainable Development Goals by 2030, there is a need for more effective policy to reduce food insecurity in low-income and lower-middle-income countries (LMIC). Measuring progress towards these goals requires reliable indicators of food security in these countries. Routinely conducted household consumption and expenditure surveys (HCES) provide potentially valuable and nationally representative data sets for this purpose. The present study aimed to assess methods used to determine national food security status using proxy measures from HCES data in LMIC globally. A scoping literature review was conducted using electronic databases. Of the 929 abstracts identified, a total of twenty articles were reviewed against strict inclusion and exclusion criteria and included for further analysis. Fourteen LMIC globally were represented in the twenty articles. The simplest metric used to indicate food insecurity compared household food expenditure against a level of expenditure considered to be below the poverty line. Data on acquisition of food was commonly converted to available energy for the household using local food composition tables and expressed as a proportion of household total energy requirements. Dietary diversity was also assessed in some studies as well as experience of food insecurity. The review demonstrated that routinely collected HCES data sets provide a useful resource for the measurement of household food security in often resource-limited LMIC. Standardisation of methods used to assess food security is needed to allow for more useful comparisons between countries, as well as to assess temporal trends.


Surface Engineered Protein Nanoparticles With Hyaluronic Acid Based Multilayers For Targeted Delivery Of Anticancer Agents.
Layer-by-layer (LbL) technique was employed to modify the surface of doxorubicin (Dox)-loaded bovine serum albumin (BSA) nanoparticles using hyaluronic acid (HA) to enable targeted delivery to overexpressed CD44 receptors in metastatic breast cancer cells. LbL technique offers a versatile approach to modify the surface of colloidal nanoparticles without any covalent modification. Dox-loaded BSA (Dox Ab) nanoparticles optimized for their size, zeta potential, and drug encapsulation efficiency were prepared by modified desolvation technique. The cellular uptake and cytotoxicity of the LbL coated Dox Ab nanoparticles were analyzed in CD44 overexpressing breast cancer cell line MDA-MB-231. Nanoparticles with HA as the final layer (Dox Ab HA) showed maximum cellular uptake in MDA-MB-231 cells owing to the CD44 receptor-mediated endocytosis and hence, exhibited more cytotoxicity as compared to free Dox. Further, luciferase-transfected MDA-MB-231 cells were used to induce tumor in BALB/c female nude mice to enable whole body tumor imaging. The mice were imaged before and after Dox treatment to visualize the tumor growth. The in vivo biodistribution of Dox Ab HA nanoparticles in nude mice showed maximum accumulation in tumor, and importantly, better tumor reduction in comparison with free Dox, thus paving the way for improved drug delivery into tumors.


Efficient and accurate fragmentation methods.
Conspectus Three novel fragmentation methods that are available in the electronic structure program GAMESS (general atomic and molecular electronic structure system) are discussed in this Account. The fragment molecular orbital (FMO) method can be combined with any electronic structure method to perform accurate calculations on large molecular species with no reliance on capping atoms or empirical parameters. The FMO method is highly scalable and can take advantage of massively parallel computer systems. For example, the method has been shown to scale nearly linearly on up to 131 000 processor cores for calculations on large water clusters. There have been many applications of the FMO method to large molecular clusters, to biomolecules (e.g., proteins), and to materials that are used as heterogeneous catalysts. The effective fragment potential (EFP) method is a model potential approach that is fully derived from first principles and has no empirically fitted parameters. Consequently, an EFP can be generated for any molecule by a simple preparatory GAMESS calculation. The EFP method provides accurate descriptions of all types of intermolecular interactions, including Coulombic interactions, polarization/induction, exchange repulsion, dispersion, and charge transfer. The EFP method has been applied successfully to the study of liquid water, π-stacking in substituted benzenes and in DNA base pairs, solvent effects on positive and negative ions, electronic spectra and dynamics, non-adiabatic phenomena in electronic excited states, and nonlinear excited state properties. The effective fragment molecular orbital (EFMO) method is a merger of the FMO and EFP methods, in which interfragment interactions are described by the EFP potential, rather than the less accurate electrostatic potential. The use of EFP in this manner facilitates the use of a smaller value for the distance cut-off (Rcut). Rcut determines the distance at which EFP interactions replace fully quantum mechanical calculations on fragment-fragment (dimer) interactions. The EFMO method is both more accurate and more computationally efficient than the most commonly used FMO implementation (FMO2), in which all dimers are explicitly included in the calculation. While the FMO2 method itself does not incorporate three-body interactions, such interactions are included in the EFMO method via the EFP self-consistent induction term. Several applications (ranging from clusters to proteins) of the three methods are discussed to demonstrate their efficacy. The EFMO method will be especially exciting once the analytic gradients have been completed, because this will allow geometry optimizations, the prediction of vibrational spectra, reaction path following, and molecular dynamics simulations using the method.


Deploying and sharing U-Compare workflows as web services.
U-Compare is a text mining platform that allows the construction, evaluation and comparison of text mining workflows. U-Compare contains a large library of components that are tuned to the biomedical domain. Users can rapidly develop biomedical text mining workflows by mixing and matching U-Compare's components. Workflows developed using U-Compare can be exported and sent to other users who, in turn, can import and re-use them. However, the resulting workflows are standalone applications, i.e., software tools that run and are accessible only via a local machine, and that can only be run with the U-Compare platform. We address the above issues by extending U-Compare to convert standalone workflows into web services automatically, via a two-click process. The resulting web services can be registered on a central server and made publicly available. Alternatively, users can make web services available on their own servers, after installing the web application framework, which is part of the extension to U-Compare. We have performed a user-oriented evaluation of the proposed extension, by asking users who have tested the enhanced functionality of U-Compare to complete questionnaires that assess its functionality, reliability, usability, efficiency and maintainability. The results obtained reveal that the new functionality is well received by users. The web services produced by U-Compare are built on top of open standards, i.e., REST and SOAP protocols, and therefore, they are decoupled from the underlying platform. Exported workflows can be integrated with any application that supports these open standards. We demonstrate how the newly extended U-Compare enhances the cross-platform interoperability of workflows, by seamlessly importing a number of text mining workflow web services exported from U-Compare into Taverna, i.e., a generic scientific workflow construction platform.


Eosin Y photoredox catalyzed net redox neutral reaction for regiospecific annulation to 3-sulfonylindoles via anion oxidation of sodium sulfinate salts.
An eosin Y photoredox catalyzed net redox neutral process for 3-sulfonylindoles via the anionic oxidation of sodium sulfinate salts and its radical cascade cyclization with 2-alkynyl-azidoarenes was developed with visible light as a mediator. The reaction offers metal and oxidant/reductant free, visible light mediated vicinal sulfonamination of alkynes to 2-aryl/alkyl-3-sulfonylindoles and proceeds via the generation of a sulfur-centered radical through direct oxidation of the sulfinate anion by an excited photocatalyst with a reductive quenching cycle. The mild conditions, use of an organic dye as photo-catalyst, bench stability and easily accessible starting materials make the present approach green and attractive.


Screening for hereditary angioedema (HAE) at 13 emergency centers in Osaka, Japan: A prospective observational study.
Hereditary angioedema (HAE) with deficiency of C1 inhibitor (C1-INH) is an autosomal-dominant disease characterized by recurrent episodes of potentially life-threatening angioedema. The objective is to study the incidence of HAE among patients who visit the emergency department.This was a 3-year prospective observational screening study involving 13 urban tertiary emergency centers in Osaka prefecture, Japan. Patients were included if they met the following criteria: unexplained edema of the body, upper airway obstruction accompanied by edema, anaphylaxis, acute abdomen with intestinal edema (including ileus and acute pancreatitis), or asthma attack. C1-INH activity and C4 level were measured at the time of emergency department admission during the period between July 2011 and June 2014.This study comprised 66 patients with a median age of 54.0 (IQR: 37.5-68.3) years. Three patients were newly diagnosed as having HAE, and 1 patient had already been diagnosed as having HAE. C1-INH activity levels of the patients with HAE were below the detection limit (<25%), whereas those of non-HAE patients (n = 62) were 106% (IQR: 85.5%-127.0%) (normal range, 70%-130%). The median level of C4 was significantly lower in the patients with HAE compared with those without HAE (1.2 [IQR: 1-3] mg/dL vs 22 [IQR: 16.5-29.5] mg/dL, P < 0.01) (normal range, 17-45 mg/dL).Three patients with undiagnosed HAE were diagnosed as having HAE in the emergency department during the 3-year period. If patients have signs and symptoms suspicious of HAE, the levels of C1-INH activity and C4 should be measured.


Mcl-1 correlates with reduced apoptosis in neutrophils from patients with sepsis.
Reduction of PMN apoptosis during sepsis contributes to the pathogenesis of multiple organ failure. Differential expression of Bcl-2 proteins, which participate in apoptosis regulation, may be responsible for the dysbalanced apoptosis seen in neutrophils from septic patients. In this study, expression of Mcl-1, Bid, Bcl-2, and Bax were investigated in septic neutrophils. PMN (1 x 10(6)/mL) from septic patients (n = 16) or healthy volunteers (n = 10) were incubated with either lipopolysaccharide (1 microg/mL), agonistic CD95 antibody (100 ng/mL), or medium for 16 hours. Apoptosis was quantified in FACS after propidium iodine staining. Mcl-1, Bid, Bcl-2, and Bax mRNA expression was detected by reverse transcriptase-polymerase chain reaction and protein determined by Western blot analysis. Spontaneous apoptosis was significantly reduced in PMN from septic patients (28.8% versus 64.0% in controls). Mcl-1 protein levels decreased in patients after 16 hours but remained stable in controls. Mcl-1 mRNA was found in freshly isolated PMN from controls and patients but remained elevated only in patients. Bid protein level decreased significantly in control PMN undergoing apoptosis but differences were less prominent in septic patients. Bid mRNA was detected only in freshly isolated PMN. No Bcl-2 mRNA or protein was detected in neutrophils from patients or controls, and detectable Bax protein and mRNA levels remained unchanged in all samples. Alterations of Bid and Mcl-1 protein in neutrophils may reflect the level of apoptosis. The upregulation of Mcl-1 mRNA in patients with sepsis suggests an active role for Mcl-1 in regulation of apoptosis during sepsis; Bax remains unchanged.