Descriptive epidemiology of injury cases: findings from a pilot injury surveillance system in Abu Dhabi.
Considering the high burden of injuries, the Health Authority-Abu Dhabi developed a draft electronic and paper-based injury and poisoning notification system (IPNS) to generate better data on the nature and severity of injuries. The pilot testing and evaluation of IPNS was conducted with the specific objectives to (1) identify the characteristics of injury cases, (2) explore potential risk factors, (3) illustrate the nature and type of data, and (4) the working mechanism of data collection. Data were collected from selected hospitals on patient demographics, injury information and clinical assessment. Descriptive, bivariate and multivariate analyses were conducted. Of 4226 injury cases, nearly three-fourths were male, majority were non-UAE nationals, and the mean age was 21.9. Multivariate findings suggested that compared to UAE nationals, non-UAE nationals were 27% more likely to experience fatal, severe or moderate injuries (p = 0.01). Individuals with health insurance were 31% less likely to suffer a fatal, severe or moderate injury compared to those having no health insurance (p < 0.001). This is the first systematically standardised collection of injury data across three facilities in Abu Dhabi, and provides initial information on characteristics and injury risk factors that will help identify the need for evidence-based intervention for injury prevention and control.


Role of endogenous carbon monoxide in the control of breathing in zebrafish (Danio rerio).
Carbon monoxide (CO) is a gaseous signaling molecule and is produced in vivo from the intracellular breakdown of heme via the heme oxygenase (HO) family of enzymes. In this study we investigated the role of the HO-1/CO system in the control of ventilation in zebrafish, Danio rerio Immunohistochemistry revealed the presence of HO-1 in the chemoreceptive neuroepithelial cells (NECs) of larvae (4 days postfertilization) and adults, indicating the potential for endogenous CO production in the NECs. Hypoxia (20 min, water Po2 of 30 mmHg) caused a significant increase in HO-1 activity in whole larvae and in the gills of adult fish. Zebrafish with reduced HO-1 activity (via HO-1 knockdown in larvae or zinc protoporphyrin IX treatment in adults) exhibited increased ventilation frequency (Vf) under normoxic but not hypoxic conditions. The addition of exogenous CO restored resting Vf in fish with diminished CO production, and in some cases (e.g., hypoxic sham larvae) CO modestly reduced Vf below resting levels. Larval fish were treated with phenylhydrazine (PHZ) to eliminate the potential confounding effects of CO-hemoglobin interactions that might influence ventilation. PHZ treatment did not cause changes in Vf of normoxic larvae, and the addition of CO to PHZ-exposed larvae resulted in a significant decrease in sham and HO-1-deficient fish under normoxic conditions. This study demonstrates for the first time that CO plays an inhibitory role in the control of breathing in larval and adult zebrafish.


Exendin-4, a GLP-1 receptor agonist, attenuates prostate cancer growth.
Recently, pleiotropic benefits of incretin therapy beyond glycemic control have been reported. Although cancer is one of the main causes of death in diabetic patients, few reports describe the anticancer effects of incretin. Here, we examined the effect of the incretin drug exendin (Ex)-4, a GLP-1 receptor (GLP-1R) agonist, on prostate cancer. In human prostate cancer tissue obtained from patients after they had undergone radical prostatectomy, GLP-1R expression colocalized with P504S, a marker of prostate cancer. In in vitro experiments, Ex-4 significantly decreased the proliferation of the prostate cancer cell lines LNCap, PC3, and DU145, but not that of ALVA-41. This antiproliferative effect depended on GLP-1R expression. In accordance with the abundant expression of GLP-1R in LNCap cells, a GLP-1R antagonist or GLP-1R knockdown with small interfering RNA abolished the inhibitory effect of Ex-4 on cell proliferation. Although Ex-4 had no effect on either androgen receptor activation or apoptosis, it decreased extracellular signal-regulated kinase (ERK)-mitogen-activated protein kinase (MAPK) phosphorylation in LNCap cells. Importantly, Ex-4 attenuated in vivo prostate cancer growth induced by transplantation of LNCap cells into athymic mice and significantly reduced the tumor expression of P504S, Ki67, and phosphorylated ERK-MAPK. These data suggest that Ex-4 attenuates prostate cancer growth through the inhibition of ERK-MAPK activation.


Fusobacteria modulate oral carcinogenesis and promote cancer progression.
Background: Evidence suggest periodontal bacterial infection can contribute to oral cancer initiation and progression. Aim: To investigate the effects of periodontal bacteria on oral cancer cell behavior using a cell-based system and a mouse carcinogenesis model. Methods: Oral cancer cell lines were polyinfected with four periodontal bacteria. Cytokine levels and relative changes in oncogene mRNA expression were determined post-infection. Oral tumours in mice induced by 4-nitroquinoline-1-oxide (4NQO) were compared with and without administrating periodontal bacteria. Results: Polyinfected oral cancer cells had upregulated MMP1, MMP9, and IL-8. The expression of cell survival markers MYC, JAK1, and STAT3 and epithelial-mesenchymal transition markers ZEB1 and TGF-β were also significantly elevated. Monoinfections showed F. nucleatum alone had comparable or greater effects than the four bacteria together. Fusobacterial culture supernatant, primarily LPS, was sufficient to induce IL-8 secretion, demonstrating that direct contact of live Fusobacteria with cancer cells might not be required to exert changes in cancer cell behaviour. In the 4NQO-induced oral tumour model, mice infected with bacteria developed significantly larger and more numerous lesions compared to those not infected. Conclusion: This study demonstrated that Fusobacteria could potentially enhance cancer cell invasiveness, survival, and EMT when presented in the oral tumour microenvironment. Abbreviations: 4NQO, 4-nitroquinoline-1-oxide; ELISA, enzyme-linked immunosorbent assay; EMT, epithelial-mesenchymal transition; IL-8, interleukin-8; JAK1, Janus kinase 1; LPS, lipopolysaccharide; MMP, matrix metalloproteinase; OSCCs, oral squamous cell carcinomas; PK, proteinase K; PMB, Polymyxin B; qRT-PCR, quantitative real-time polymerase chain reaction; STAT3, signal transducer and activator of transcription 3; TGF-β, transforming growth factor beta; ZEB1, zinc finger E-Box binding homeobox 1.


Sodium Fate after Sodium Bicarbonate Infusion: Influence of Altered Acid-Base Status.
We have previously investigated the fate of administered bicarbonate infused as a hypertonic solution in animals with each of the 4 chronic acid-base disorders. Those studies did not address the fate of sodium, the coadministered cation. We examined baseline total body water (TBW), Na+ space, HCO3- space, and urinary sodium and bicarbonate excretion after acute hypertonic NaHCO3 infusion (1-N solution, 5 mmol/kg body weight) in dogs with each of the 4 chronic acid-base disorders. Observations were made at 30, 60, and 90 min postinfusion. Retained sodium that remains osmotically active distributes in an apparent space that approximates TBW. Na+ space that exceeds TBW uncovers nonosmotic sodium storage. Na+ space approximated TBW at all times in normal and hyperbicarbonatemic animals (metabolic alkalosis and respiratory acidosis), but exceeded TBW by ~30% in hypobicarbonatemic animals (metabolic acidosis and respiratory alkalosis). Such osmotic inactivation was detected at 30 min and remained stable. The pooled data revealed that Na+ space corrected for TBW was independent of the initial blood pH but correlated with initial extracellular bicarbonate concentration (y = -0.01x + 1.4, p= 0.002). The fate of administered sodium and bicarbonate (internal distribution and urinary excretion) was closely linked. This study demonstrates that hypobicarbonatemic animals have a Na+ space that exceeds TBW after an acute infusion of hypertonic NaHCO3 indicating osmotic inactivation of a fraction of retained sodium. In addition to an expanded Na+ space, these animals have a larger HCO3- space compared with hyperbicarbonatemic animals. Both phenomena appear to reflect the wider range of titration of nonbicarbonate buffers (Δ pH) occurring during NaHCO3- loading whenever initial [HCO3-]e is low. The data indicate that the fate of administered bicarbonate drives the internal distribution and the external disposal of sodium, the co-administered cation, and is responsible for the early, but non-progressive, osmotic inactivation of a fraction of the retained sodium.


Environmental and demographic correlates of bicycling.
The present study examined correlates of bicycle ownership and bicycling frequency, and projected increases in cycling if perceived safety from cars was improved. Participants were 1780 adults aged 20-65 recruited from the Seattle, Washington and Baltimore, Maryland regions (48% female; 25% ethnic/racial minority) and studied in 2002-2005. Bicycling outcomes were assessed by survey. Multivariable models were conducted to examine demographic and built environment correlates of bicycling outcomes. About 71% of the sample owned bicycles, but 60% of those did not report cycling. Among bicycle owners, frequency of riding was greater among young, male, White, educated, and lean subgroups. Neighborhood walkability measures within 1 km were not consistently related to bicycling. For the whole sample, bicycling at least once per week was projected to increase from 9% to 39% if bicycling was safe from cars. Ethnic-racial minority groups and those in the least safe neighborhoods for bicycling had greater projected increases in cycling if safety from traffic was improved. Implementing measures to improve bicyclists' safety from cars would primarily benefit racial-ethnic groups who cycle less but have higher rates of chronic diseases, as well as those who currently feel least safe bicycling.


Micromachined ultrasonic capacitance transducers for immersion applications.
Investigations into the characteristics of water-coupled ultrasonic capacitance transducers have been undertaken for a range of transducer configurations. The radiated fields have been scanned in water using a miniature hydrophone detector, and the results compared to theory based on a plane piston approach. Micromachined backplates in conjunction with thin Mylar and mica membranes have been investigated, together with aperture modifications such as an annulus and Fresnel zone plate. The measured results agree well with theory, thus demonstrating that wideband predictable performance (>8 MHz) is obtainable with such transducers. Additionally, pulse-echo C-scans of a Plexiglas plate containing an artificial defect have been undertaken with the capacitance transducer, in order to demonstrate that the capacitance transducer has sufficient sensitivity to allow routine nondestructive testing within immersion applications.


Electrochemistry meets polymer physics: polymerized ionic liquids on an electrified electrode.
Polymeric ionic liquids are emerging polyelectrolyte materials for modern electrochemical applications. In this paper, we propose a self-consistent field theory of a polymeric ionic liquid on a charged conductive electrode. Taking into account the conformational entropy of rather long polymerized cations within the Lifshitz theory and electrostatic and excluded volume interactions of ionic species within the mean-field approximation, we obtain a system of self-consistent field equations for the local electrostatic potential and average concentrations of monomeric units and counterions. We solve these equations in the linear approximation for the cases of a point-like charge and a flat infinite uniformly charged electrode immersed in a polymeric ionic liquid and derive analytical expressions for local ionic concentrations and electrostatic potential, and derive an analytical expression for the linear differential capacitance of the electric double layer. We also find a numerical solution to the self-consistent field equations for two types of boundary conditions for the local polymer concentration on the electrode, corresponding to the cases of the specific adsorption absence (indifferent surface) and strong short-range repulsion of the monomeric units near the charged surface (hard wall case). For both cases, we investigate the behavior of differential capacitance as a function of applied voltage for a pure polymeric ionic liquid and a polymeric ionic liquid dissolved in a polar organic solvent. We observe that the differential capacitance profile shape is strongly sensitive to the adopted boundary condition for the local polymer concentration on the electrode.


Postoperative malnutrition in Duchenne muscular dystrophy.
Dysphagia and aspiration seem to be rare in Duchenne muscular dystrophy, but cachexia can be associated with early death. Commonly, weight loss can be attributed to inadequate caloric intake caused by loss of ability to self-feed and/or fatigue. Our objective was to determine whether scoliosis repair is associated with malnutrition. A retrospective chart review was undertaken of patients with Duchenne muscular dystrophy, including those who underwent operative repair of scoliosis. We identified nine boys who lost > 5% body weight within 12 months of surgery. Eight patients who gained weight after surgery and eight patients of comparable age who had no surgery served as control subjects. All patients had no change in biceps strength after surgery, but those who lost weight were unable to self-feed. We found that weight loss after surgery was associated with loss of self-feeding. We conclude that pre- and postoperative management of patients with Duchenne muscular dystrophy should include feeding evaluation and determination of postural changes.


Niche specificity of ammonia-oxidizing archaeal and bacterial communities in a freshwater wetland receiving municipal wastewater in Daqing, Northeast China.
Ecophysiological differences between ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) enable them to adapt to different niches in complex freshwater wetland ecosystems. The community characters of AOA and AOB in the different niches in a freshwater wetland receiving municipal wastewater, as well as the physicochemical parameters of sediment/soil samples, were investigated in this study. AOA community structures varied and separated from each other among four different niches. Wetland vegetation including aquatic macrophytes and terrestrial plants affected the AOA community composition but less for AOB, whereas sediment depths might contribute to the AOB community shift. The diversity of AOA communities was higher than that of AOB across all four niches. Archaeal and bacterial amoA genes (encoding for the alpha-subunit of ammonia monooxygenases) were most diverse in the dry-land niche, indicating O2 availability might favor ammonia oxidation. The majority of AOA amoA sequences belonged to the Soil/sediment Cluster B in the freshwater wetland ecosystems, while the dominant AOB amoA sequences were affiliated with Nitrosospira-like cluster. In the Nitrosospira-like cluster, AOB amoA gene sequences affiliated with the uncultured ammonia-oxidizing beta-proteobacteria constituted the largest portion (99%). Moreover, independent methods for phylogenetic tree analysis supported high parsimony bootstrap values. As a consequence, it is proposed that Nitrosospira-like amoA gene sequences recovered in this study represent a potentially novel cluster, grouping with the sequences from Gulf of Mexico deposited in the public databases.


Family Assisted Contingency Management within the Context of Evidence-Supported Treatment for Child Neglect and Drug Abuse.
Contingency management (CM) has extensively been shown to be effective in reducing substance use disorders, but its effects in reducing child maltreatment have yet to be determined. The current study provides preliminary support for the utilization of an innovative family-assisted CM component in 18 mothers who were referred to an evidence-supported behavioral treatment for concurrent child neglect and drug abuse by Child Protective Service caseworkers. In the examined CM, participants were invited to indicate from a list of common actions incompatible with child neglect (i.e. positive parenting actions), the extent to which these actions had been experienced utilizing a 3-point scale (almost never, sometimes, almost always). For each item that was indicated to be almost never or sometimes experienced, the participants were queried to indicate if the neglect incompatible action should be targeted as a therapeutic goal. Contingencies were subsequently established in which the participants were rewarded by involved family members for their completion of therapeutic goals. At baseline, results indicated that there was a negative association between the number of neglect incompatible parenting actions that were infrequently experienced and child abuse potential. A hierarchical multiple regression analysis showed that the number of neglect incompatible actions targeted as therapeutic goals at baseline, but not the number of positive parenting actions experienced infrequently at baseline, predicted reduced child maltreatment potential following treatment. These findings suggest the examined CM may assist evidence supported behavioral treatment specific to child neglect and drug abuse.


ErbB2 upregulates the Na+,HCO3(-)-cotransporter NBCn1/SLC4A7 in human breast cancer cells via Akt, ERK, Src, and Kruppel-like factor 4.
Misregulation of acid-base transport plays central roles in cancer development. We previously demonstrated the strong up-regulation of the Na(+),HCO3(-) cotransporter NBCn1 (SLC4A7) in MCF-7 breast cancer cells by a truncated, constitutively active ErbB2 (HER2) receptor, ΔNErbB2, and showed that NBCn1 expression and activity are increased in breast cancer tissue from patients. Here, we present the first in-depth characterization of an SLC4A7 promoter and identify its minimal ΔNErbB2-sensitive region. Inhibition or siRNA-mediated knockdown of PI3K, Akt1, ERK1/2, or Src decreased the NBCn1 protein level in ΔNErbB2-expressing MCF-7 cells by ~50, 60, 30 and 35%, respectively. Further, knockdown of the transcription factor Krüppel-like factor 4 (KLF4) reduced NBCn1 protein expression by ~40%, and KLF4 overexpression increased NBCn1 expression by 50-80%. In contrast, knockdown of the closely related transcription factor specificity protein 1 (Sp1) or transfection with dominant-negative Sp1 increased NBCn1 expression by ~35 and ~50%, respectively. NBCn1 expression was also increased by stimulation of full-length ErbB1, -2, and -3 receptors in SKBr3 cells (1.5- and 2-fold by NRG1 or EGF, respectively) or after their exogenous expression in MCF-7 cells. Finally, stimulation with NRG1 or EGF more than doubled acid extrusion capacity in SKBr3 cells. In conclusion, NBCn1 is strongly upregulated by ErbB receptor signaling in a manner involving opposite effects of KLF4 and Sp1, transcription factors with central roles in cancer development. ErbB-induced up-regulation of NBCn1-mediated acid extrusion may play important physiological and pathophysiological roles in the breast epithelium and other tissues with high ErbB receptor levels.


Amplified energy transfer in conjugated polymer nanoparticle tags and sensors.
Nanoparticles primarily consisting of π-conjugated polymers have emerged as extraordinarily bright fluorescent tags with potential applications in biological imaging and sensing. As fluorescent tags, conjugated polymer nanoparticles possess a number of advantageous properties, such as small particle size, extraordinary fluorescence brightness, excellent photostability, and high emission rate. Exciton diffusion occurring in the nanoparticles results in amplified energy transfer, doubling the energy transfer efficiency in some cases. Amplified energy transfer has been exploited to obtain highly red-shifted emission, oxygen-sensing nanoparticles, and fluorescence photoswitching. Additional observed phenomena are attributable to amplified energy transfer in conjugated polymers, including superquenching by metal nanoparticles, and fluorescence modulation by hole polarons. This feature article presents an overview of recent investigations of optical properties and energy transfer phenomena of this relatively novel type of fluorescent nanoparticle with a viewpoint towards demanding fluorescence-based imaging and sensing applications.


Local challenge of oral mucosa with gliadin in patients with coeliac disease.
In coeliac disease, gluten-containing diet challenges over many years are sometimes required for diagnosis, especially if the initial diagnosis was equivocal. The rectal gluten challenge has been proposed to simplify coeliac disease diagnosis. We were interested in studying whether the oral mucosa could be used for local challenge with gliadin as an aid in finalizing the diagnosis of coeliac disease. The study groups consisted of 37 treated coeliac disease patients and 10 controls. The challenges on the oral mucosa were performed either supramucosally with gliadin powder (coeliac disease patients) or by submucosal injection of dissolved gliadin (10 microg/ml) (coeliac disease patients and controls). A control challenge with submucosal gliadin solvent was made in the coeliac disease patients. B and T cells, mast cells and T cell subsets were counted and HLA-DR expression was determined. Biopsies were taken from each provoked area 24 h post-challenge. A significant increase in the number of CD4+ lymphocytes in the lamina propria (observed in 27/37 patients), but a decrease in the number of mast cells was observed in treated coeliac disease patients after submucosal challenge with gliadin. Following supramucosal challenge with gliadin the counts of intraepithelial CD4+ (in 25/37 patients) and CD8+ T cells (in 27/37 patients) increased significantly and the number of CD4+ T cells in the lamina propria was also significantly increased. Control subjects were tested by submucosal gliadin challenge and no significant changes in the number of cells were observed. HLA-DR expression did not show increased positivity in coeliac disease patients on submucosal challenge. For the first time the oral mucosa has been used for immunological testing and shown to react to gliadin challenge in coeliac disease patients. Recruitment of T cells upon submucosal gliadin challenge occurred towards the lamina propria, whereas it occurred towards the epithelium in supramucosal gliadin challenge. The numbers of T cells increased in the lamina propria after submucosal challenge. The results suggest that local oral challenge with gliadin may be used as a diagnostic method in coeliac disease; however, further studies in untreated coeliac disease patients are needed to evaluate the usefulness of this method.


Joint fluid after aspiration. A disturbing factor in knee arthrography.
The amount of fluid in the knee after aspirating any effusion was investigated and averaged 6.8 ml, being independent of the presence of a lesion. Because it causes dilution of the contrast medium, it is suggested that small amounts of a relatively high-concentrated non-ionic contrast agent should be preferred for double-contrast knee arthrography.


Implementation of the WHO guideline on treatment of young infants with signs of possible serious bacterial infection when hospital referral is not feasible in rural Zaria, Nigeria: Challenges and solutions.
Bacterial infection is one of the leading causes of mortality in young infants globally. The standard practice to manage young infants with any sign of possible serious bacterial infection (PSBI) is in a hospital setting with parenteral antibiotics, which may not be feasible for majority of cases in most low resource settings. The World Health Organization developed a guideline on management of PSBI in young infants when referral is not feasible in 2016. We conducted implementation research in selected communities in Zaria Local Government Areas of Kaduna State with an estimated population of 50,000 with the aim of understanding how to implement the WHO PSBI treatment guideline to achieve high coverage with low case fatality and treatment failure rates. Implementation was within the programmatic settings using existing health structure. We conducted policy dialogue with decision makers to adapt the recommendations to their social, cultural and programmatic context in Nigeria, held orientation meetings with program managers, built capacity of the health workers and supported the implementation within the health system. We supported a non-government organization to conduct community sensitization to promote care seeking and adherence to treatment advice. The research team collected data systematically on all young infants identified to have PSBI, the treatment they received and the clinical outcome. Between April 2016 and March 2017, we identified 347 young infants up to 2 months of age with signs of PSBI who received treatment either as an outpatient or in a hospital among 2,154 births in the study population. The coverage of PSBI treatment in the study area was 95.5% assuming that 10% of all births have an episode of PSBI in the first two months of life. Most (89%) sick young infants with PSBI were identified by the community-oriented resource persons and sent to the Primary Health Care Centres (PHCs). Most families (97%) refused referral and were treated at a primary health care centre on outpatient basis. There were 12 deaths (3.5%) and 17 non-death treatment failures (4.9%) in 343 infants in whom an outcome could be ascertained. While non-death treatment failure rate was highest in 0-6-day infants with fast breathing (14.4%), case fatality was highest in those with signs of critical illness (20%). We have demonstrated that outpatient treatment strategy for young infants with PSBI when referral is not feasible is implementable within the programmatic settings, achieving very high population coverage and relatively low treatment failure and case fatality rates. Implementation at scale will require government's commitment to strengthen the health system with trained, motivated health care providers and necessary commodities.


Customized Consensus Spectral Library Building for Untargeted Quantitative Metabolomics Analysis with Data Independent Acquisition Mass Spectrometry and MetaboDIA Workflow.
Data independent acquisition-mass spectrometry (DIA-MS) coupled with liquid chromatography is a promising approach for rapid, automatic sampling of MS/MS data in untargeted metabolomics. However, wide isolation windows in DIA-MS generate MS/MS spectra containing a mixed population of fragment ions together with their precursor ions. This precursor-fragment ion map in a comprehensive MS/MS spectral library is crucial for relative quantification of fragment ions uniquely representative of each precursor ion. However, existing reference libraries are not sufficient for this purpose since the fragmentation patterns of small molecules can vary in different instrument setups. Here we developed a bioinformatics workflow called MetaboDIA to build customized MS/MS spectral libraries using a user's own data dependent acquisition (DDA) data and to perform MS/MS-based quantification with DIA data, thus complementing conventional MS1-based quantification. MetaboDIA also allows users to build a spectral library directly from DIA data in studies of a large sample size. Using a marine algae data set, we show that quantification of fragment ions extracted with a customized MS/MS library can provide as reliable quantitative data as the direct quantification of precursor ions based on MS1 data. To test its applicability in complex samples, we applied MetaboDIA to a clinical serum metabolomics data set, where we built a DDA-based spectral library containing consensus spectra for 1829 compounds. We performed fragment ion quantification using DIA data using this library, yielding sensitive differential expression analysis.


Differential area profiles: decomposition properties and efficient computation.
Differential area profiles (DAPs) are point-based multiscale descriptors used in pattern analysis and image segmentation. They are defined through sets of size-based connected morphological filters that constitute a joint area opening top-hat and area closing bottom-hat scale-space of the input image. The work presented in this paper explores the properties of this image decomposition through sets of area zones. An area zone defines a single plane of the DAP vector field and contains all the peak components of the input image, whose size is between the zone's attribute extrema. Area zones can be computed efficiently from hierarchical image representation structures, in a way similar to regular attribute filters. Operations on the DAP vector field can then be computed without the need for exporting it first, and an example with the leveling-like convex/concave segmentation scheme is given. This is referred to as the one-pass method and it is demonstrated on the Max-Tree structure. Its computational performance is tested and compared against conventional means for computing differential profiles, relying on iterative application of area openings and closings. Applications making use of the area zone decomposition are demonstrated in problems related to remote sensing and medical image analysis.


Microbial keratitis following lamellar keratoplasty.
To determine the predisposing factors, etiologic agents, and clinical and visual outcomes in infectious keratitis following lamellar keratoplasty (LK). One hundred thirty-five eyes (135 patients) that had undergone LK were retrospectively analyzed for the occurrence of infectious keratitis following LK. The parameters evaluated were predisposing factors, seasonal variation, indications and type of LK, time interval between LK and infection, site and depth of infection, etiologic organisms, type of treatment, outcome in terms of graft status, secondary surgery, visual acuity, and the donor tissue profile. The incidence of infectious keratitis following LK was 11.11%. The most significant predisposing factor was persistent epithelial defect (3 eyes) and suture abscesses (3 eyes). Most cases occurred between May and August (9/15). Twelve cases developed infection within 2 weeks of surgery (80%). Seven cases (7/15) occurred with onlay grafts, 6 with inlay grafts, and 2 with large-diameter LK. Cultures of corneal scrapings were positive in 11 eyes (73.3%), and the most common isolated organism was coagulase-negative Staphylococcus (CNS). Only 2 eyes responded to medical therapy, and graft sloughing occurred in 9 cases. Six eyes underwent penetrating keratoplasty to either salvage the integrity of globe or for visual rehabilitation of cases where infection resulted in corneal opacity. Infections after LK may not be amenable to antimicrobial therapy and may necessitate the removal of the graft or a therapeutic penetrating keratoplasty.


Identification of the isoamylase 3 gene in common wheat and its expression profile during the grain-filling period.
In higher plants, isoamylase-type starch debranching enzyme catalyzes the α-1,6-glucosidic linkages of glycogen and phytoglycogen. We cloned an isoamylase-type starch debranching enzyme ISA3 cDNA sequence (2883 bp), designated as TaISA3, from common wheat (Triticum aestivum), using the rapid amplification of cDNA ends method. The open reading frame of TaISA3 was found to have 2331 bp, and its deduced amino acid sequence was found to share high similarity with those of other gramineous plant ISA3 proteins. It contains a putative transit peptide (68 amino acids), N-terminus domain (107 amino acids), and a catalytic domain (173 amino acids). We extracted the expressed TaISA3 protein from Escherichia coli (BL21), and measured starch isoamylase activity. During the wheat grain-filling period, transcripts of the TaISA3 gene reached a maximum level at the early developmental stage, then declined, and increased again near the final maturation stage of the grain. We confirm that the ISA3 gene is present in common wheat; it appears to play a role in starch synthesis during early and late stages of the grain-filling period.


Microbulbifer gwangyangensis sp. nov. and Microbulbifer pacificus sp. nov., isolated from marine environments.
Two novel Gram-stain-negative, chemoheterotrophic and strictly aerobic bacteria, strains GY2(T) and SPO729(T), were isolated from a tidal flat at Gwangyang Bay in Korea and a marine sponge sample from the Pacific Ocean, respectively. The two strains were halotolerant, catalase- and oxidase-positive, and non-motile rods. Optimum temperature and pH for growth of both strains were observed to be 35 °C and pH 7.0-7.5, but optimum salinity for strain SPO729(T) [2-3 % (w/v)] was slightly higher than that for strain GY2(T) (1-2 %). The major cellular fatty acids of both strains were C16 : 0, iso-C15 : 0, iso-C17 : 0, iso-C17 : 1ω9c, C18 : 1ω7c, iso-C11 : 0 and iso-C11 : 0 3-OH. The genomic DNA G+C contents of strains GY2(T) and SPO729(T) were 55.1 and 57.9 mol%, respectively, and ubiquinone 8 (Q-8) was detected as the sole respiratory quinone from the two strains. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains GY2(T) and SPO729(T) formed tight phyletic lineages with members of the genus Microbulbifer. Strain GY2(T) was closely related to Microbulbifer okinawensis ABABA23(T) (98.2 %), strain SPO729(T) (98.0 %) and Microbulbifer donghaiensis CN85(T) (97.0 %); strain SPO729(T) was closely related to M. okinawensis ABABA23(T) (98.3 %) and M. donghaiensis CN85(T) (98.2 %). The DNA-DNA relatedness values of strain GY2(T) with M. okinawensis ABABA23(T), strain SPO729(T) and M. donghaiensis CN85(T) were 40.0±2.1 %, 13.1±3.9 % and 16.2±5.8 %, respectively, whereas those of strain SPO729(T) with M. okinawensis ABABA23(T) and M. donghaiensis CN85(T) were 48.0±4.0 % and 34.6±9.3 %, respectively. On the basis of phenotypic and molecular features, it is concluded that the two strains GY2(T) and SPO729(T) represent two novel species of the genus Microbulbifer, for which the names Microbulbifer gwangyangensis sp. nov. and Microbulbifer pacificus are proposed; the type strains are GY2(T) ( = KACC 16189(T) = JCM 17800(T)) and SPO729(T) ( = KCCM 42667(T) = JCM 14507(T)), respectively.


Context-Aware Reviewer Assignment for Trust Enhanced Peer Review.
Reviewer assignment is critical to peer review systems, such as peer-reviewed research conferences or peer-reviewed funding applications, and its effectiveness is a deep concern of all academics. However, there are some problems in existing peer review systems during reviewer assignment. For example, some of the reviewers are much more stringent than others, leading to an unfair final decision, i.e., some submissions (i.e., papers or applications) with better quality are rejected. In this paper, we propose a context-aware reviewer assignment for trust enhanced peer review. More specifically, in our approach, we first consider the research area specific expertise of reviewers, and the institution relevance and co-authorship between reviewers and authors, so that reviewers with the right expertise are assigned to the corresponding submissions without potential conflict of interest. In addition, we propose a novel cross-assignment paradigm, and reviewers are cross-assigned in order to avoid assigning a group of stringent reviewers or a group of lenient reviewers to the same submission. More importantly, on top of them, we propose an academic CONtext-aware expertise relevanCe oriEnted Reviewer cross-assignmenT approach (CONCERT), which aims to effectively estimate the "true" ratings of submissions based on the ratings from all reviewers, even though no prior knowledge exists about the distribution of stringent reviewers and lenient reviewers. The experiments illustrate that compared with existing approaches, our proposed CONCERT approach can less likely assign more than one stringent reviewers or lenient reviewers to a submission simultaneously and significantly reduce the influence of ratings from stringent reviewers and lenient reviewers, leading to trust enhanced peer review and selection, no matter what kind of distributions of stringent reviewers and lenient reviewers are.


High-resolution melting analysis of BLU methylation levels in gastric, colorectal, and pancreatic cancers.
BLU was recently characterized as a novel tumor suppressor gene (TSG), and was epigenetically silenced in some tumor cell lines and primary tumor samples. High-resolution melting (HRM) analysis has been used as a novel tool for analysis of promoter methylation. We used HRM analysis to detect the methylation levels of BLU gene in 100 gastric, 100 colorectal, and 70 pancreatic cancers, and also in an equal number of adjacent normal tissues for all. The frequency of BLU methylation in all three types of cancer was significantly higher than that in normal tissues. And the expression levels of BLU were inversely correlated with methylation levels.


Oral biopsies: methods and applications.
Biopsies are an important diagnostic tool for the diagnosis of lesions ranging from simple periapical lesions to malignancies. Planning prior to performing a biopsy is essential. It will be beneficial to the receiving pathologist in reaching a helpful and meaningful diagnosis, and therefore ultimately and more importantly, to the patient. This paper presents an updated view of biopsies and discusses some of the potential problems with biopsy technique and specimens and how to overcome them.


"Preconditioning at a distance" in the isolated rabbit heart.
Brief myocardial ischemia evokes a cardioprotective response, referred to as "ischemic preconditioning" (IP), that limits injury caused by a subsequent prolonged ischemic insult. The myocardial IP effect can be induced by ischemia of "distant" cardiac and noncardiac tissue, implicating the involvement of an as-yet-unidentified humoral trigger. If a preconditioning hormone exists, the authors hypothesize that the IP effect should be transferable, via administration of coronary effluent, from a preconditioned donor heart to a virgin non-preconditioned acceptor heart. Isolated buffer-perfused rabbit hearts were assigned to one of four treatment groups in a donor/acceptor sequence. Donor hearts underwent either three IP cycles or a matched period of uninterrupted perfusion (control donors). Coronary perfusate collected from IP and control donor hearts was reoxygenated and transfused to virgin acceptor hearts. All hearts then underwent 30 minutes of global ischemia followed by 30 minutes of reperfusion. Left ventricular developed pressure (LVDP) (the authors' index of cardioprotection) was monitored throughout the protocol by a left ventricular (LV) balloon. In donor controls, LVDP assessed at 30 minutes post-reflow was restored to only 49 +/- 5% of baseline values. Recovery of LV function was significantly enhanced in both IP donor hearts (69 +/- 4%*) and IP acceptor hearts (70 +/- 6%*) vs donor controls (*p < 0.05), while, in acceptor controls, intermediate values of LVDP (62 +/- 7%) were obtained. The IP effect can be transferred between rabbit hearts, suggesting the presence of a humoral trigger signal for distant preconditioning. Isolating this hormone may have therapeutic and diagnostic implications in the management of acute myocardial ischemia.


Practical approaches for self-monitoring of blood glucose: an Asia-Pacific perspective.
Comprehensive glycemic control is necessary to improve outcomes and avoid complications in individuals with diabetes. Self-monitoring of blood glucose (SMBG) is a key enabler of glycemic assessment, providing real-time information that complements HbA1c monitoring and supports treatment optimization. However, SMBG is under-utilized by patients and physicians within the Asia-Pacific region, because of barriers such as the cost of monitoring supplies, lack of diabetes self-management skills, or concerns about the reliability of blood glucose readings. Practice recommendations in international and regional guidelines vary widely, and may not be detailed or specific enough to guide SMBG use effectively. This contributes to uncertainty among patients and physicians about how best to utilize this tool: when and how often to test, and what action(s) to take in response to high or low readings. In developing a practical SMBG regimen, the first step is to determine the recommended SMBG frequency and intensity needed to support the chosen treatment regimen. If there are practical obstacles to monitoring, such as affordability or access, physicians should identify the most important aspects of glycemic control to target for individual patients, and modify monitoring patterns accordingly. This consensus paper proposes a selection of structured, flexible SMBG patterns that can be tailored to the clinical, educational, behavioral, and financial requirements of individuals with diabetes.


Going beyond the information given: a neural system supporting semantic interpretation.
Relating the meaning of a word to the context in which it is encountered is central to comprehension. We investigated the neural basis of this process. Subjects made decisions based on a semantic property of single nouns. The lack of sentence context created ambiguity, as nouns may have several, unrelated semantic identities. Contrasted with unambiguous decisions about each noun's sound structure, the semantic task resulted in activity in the left superior frontal gyrus (SFG), activity that was dependent on choice reaction time. This identified the left SFG as an executive component of a distributed cognitive system that relates a word's meaning to its semantic context to facilitate comprehension.


The Rising Dominance of Shigella sonnei: An Intercontinental Shift in the Etiology of Bacillary Dysentery.
Shigellosis is the major global cause of dysentery. Shigella sonnei, which has historically been more commonly isolated in developed countries, is undergoing an unprecedented expansion across industrializing regions in Asia, Latin America, and the Middle East. The precise reasons underpinning the epidemiological distribution of the various Shigella species and this global surge in S. sonnei are unclear but may be due to three major environmental pressures. First, natural passive immunization with the bacterium Plesiomonas shigelloides is hypothesized to protect populations with poor water supplies against S. sonnei. Improving the quality of drinking water supplies would, therefore, result in a reduction in P. shigelloides exposure and a subsequent reduction in environmental immunization against S. sonnei. Secondly, the ubiquitous amoeba species Acanthamoeba castellanii has been shown to phagocytize S. sonnei efficiently and symbiotically, thus allowing the bacteria access to a protected niche in which to withstand chlorination and other harsh environmental conditions in temperate countries. Finally, S. sonnei has emerged from Europe and begun to spread globally only relatively recently. A strong selective pressure from localized antimicrobial use additionally appears to have had a dramatic impact on the evolution of the S. sonnei population. We hypothesize that S. sonnei, which exhibits an exceptional ability to acquire antimicrobial resistance genes from commensal and pathogenic bacteria, has a competitive advantage over S. flexneri, particularly in areas with poorly regulated antimicrobial use. Continuing improvement in the quality of global drinking water supplies alongside the rapid development of antimicrobial resistance predicts the burden and international distribution of S. sonnei will only continue to grow. An effective vaccine against S. sonnei is overdue and may become one of our only weapons against this increasingly dominant and problematic gastrointestinal pathogen.


Axonal injury in multiple sclerosis.
The pivotal role of axons in the pathophysiology and pathogenesis of multiple sclerosis (MS) is increasingly becoming the focus of our attention. Axonal injury, considered at one time to be a late phenomenon, is now recognized as an early occurrence in the inflammatory lesions of MS. There is converging evidence from histopathologic, as well as magnetic resonance imaging and magnetic resonance spectroscopy studies, that axons play a crucial and dynamic role during the evolution of MS pathology and the development of clinical disability. It has been repeatedly demonstrated that neurologic functional impairment correlates best with axonal, rather than myelin, injury. The pathophysiology of axonal injury remains speculative. Although generally considered to be sequelae of demyelination, it is possible that axonal injury in MS is indeed a primary event. The discovery that axonal injury can be reversible has provided an impetus to institute early therapy. The finding that irreversible axonal transection occurs in early lesions has underscored now, more than ever before, the need to curtail inflammation and the need to institute early treatment with disease-modifying agents. The axon will undoubtedly remain the focus of our attention regarding research on MS now and in the future.


Functional and topographical analyses of epitopes on the hemagglutinin (VP4) of the simian rotavirus SA11.
An immunochemical analysis of the hemagglutinin (VP4) of the simian rotavirus SA11 was performed to better understand the structure and function of this molecule. Following immunization of mice with double-shelled virus particles and VP4-enriched fractions from CsCl gradients, a battery of anti-SA11 hybridomas was generated. A total of 13 clones secreting high levels of anti-VP4 monoclonal antibody (MAb) was characterized and compared with two cross-reactive anti-VP4 MAbs generated against heterologous rhesus (RRV) and porcine (OSU) rotavirus strains. These cross-reactive MAbs effectively neutralized SA11 infectivity in vitro. The epitopes recognized by these 15 MAbs were grouped into six antigenic sites on the SA11 hemagglutinin. These sites were identified following analysis of the MAbs by using a simple competitive binding enzyme-linked immunosorbent assay (ELISA) and biological assays. Three of the antigenic sites were involved in neutralization of virus infectivity in vitro. All the MAbs with neutralization activity and two nonneutralizing MAbs were able to inhibit viral hemagglutination of human erythrocytes. Competitive binding ELISA data showed a positive cooperative binding effect with some pairs of the anti-VP4 MAbs, apparently due to a conformational change induced by the binding of the first MAb. Some of the MAbs also bound better to trypsin-treated virus than to non-trypsin-treated virus. A topographic map for VP4 is proposed on the basis of the observed properties of each antigenic site.


Genome-wide identification and expression analysis of heat shock protein gene family in cassava.
Heat shock proteins are important molecular chaperones that are involved in plant growth and stress responses. However, members of the Hsp family have been poorly studied in cassava. In this study, 225 MeHsp genes were identified in the cassava genome, and their genetic structures exhibited relatively conserved features within each subfamily. The 225 MeHsp genes showed random chromosomal distribution, and at least 74 pairs of segmentally duplicated MeHsp genes. Eleven tandemly duplicated MeHsp genes were identified. Cis-element analysis revealed the importance of MeHsps in plant adaptations to the environment. The prediction of protein interactions suggested that MeHsp70-20 may play a critical regulatory role in the interactive network. Furthermore, the expression profiles of MeHsps in different tissues and cell subsets were analyzed using bulk transcriptomics and single-cell transcriptomic data. Several subfamily genes exhibited unique expression patterns in the transcriptome and were selected for detailed analysis of the single-cell transcriptome. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed the expression patterns of these genes under temperature stress, further supporting the prediction of cis-acting elements. This study provides valuable information for understanding the functional characteristics of MeHsp genes and the evolutionary relationships between MeHsps.


The retention of photosynthetic activity by senescing chloroplasts of oat leaves.
The retention of photosystems I and II and or RuDP carboxylase activity in chloroplasts isolated from the first leaves of Victory oat (Avena sativa L.) seedlings was followed as the chloroplasts senesced in darkness. Both photosystems (PS) I and II retained their full activity after 3 days at 1°C, while even after 7 days at 1°C around 80% of the activity was still present. After 3 days at 25°C, PS I lost only 20% and PS II 50% of the initial activity. Acid pH increased the rate of decay of both systems, PS II falling almost to zero after 3 days at pH 3.5 (at 25°C). The preparations were almost bacteria-free, and addition of antibiotics not only did not improve their stability, but accelerated the rates of loss of photosynthetic activity. This is held to indicate that the enzymes are undergoing some turnover even in isolated chloroplasts. If the leaves were allowed to senesce in the dark first and the chloroplasts then isolated, their photosynthetic activities had greatly decreased, showing that senescence is more rapid in situ than in isolation. Under these conditions PS I decayed more rapidly than PS II. Ribulosediphosphate carboxylase, as measured by CO2 fixation, declined more rapidly than the photosystems, though the addition of kinetin and indole-3-acetic acid somewhat decreased the rate of loss, at least for the first 24 h. When the intact (detached) leaves were held in the dark, the rate of oxygen evolution declined rapidly, but in monochromatic blue light (450 nm) at 25°C about 30% of the initial rate was retained after 72 h.


Effect of noise on the vestibular system - Vestibular evoked potential studies in rats.
Studies have shown that in order for sound to affect the vestibular end organs in the inner ear, very high intensities are required. Furthermore, in patients with noise induced hearing loss, vestibular signs, if present, are subclinical. In order to study possible auditory-vestibular interactions in a more controlled fashion, using physiological sound intensities, the present study used short latency vestibular evoked potentials (VsEPs) to impulses of angular (15,000 degrees /sec(2), risetime 1.5 msec) and linear (3-5 g, risetime 1.5 msec) acceleration were used to study the possible effects of sound on peripheral vestibular function in rats. Four different paradigms were used: a - an intense (135 dB pe SPL) click stimulus was presented 5 msec before the linear acceleration impulse and the VsEP to 128 stimuli were recorded with and without this click stimulus. There was no effect of the preceding intense click on the first wave (reflecting end organ activity) of the linear VsEP. b - 113 dB SPL white noise "masking" was presented while the VsEPs were elicited. A 10-20% reduction in the amplitude of the first VsEP wave was seen during the noise exposure, but 5 minutes after this exposure, there was almost complete recovery to pre-exposure amplitude. c - 113 dB SPL noise was presented for one hour and VsEPs were recorded within 15 minutes of cessation of the noise. The auditory nerve-brainstem-evoked response showed a temporary threshold shift while there was no effect on the VsEP. d - 113 dB SPL white noise was presented for 12 hours per day for 21 consecutive days. Auditory nerve-brainstem-evoked responses and vestibular (VsEPs) function were studied one week after the conclusion of the noise exposure. Auditory function was severely permanently depressed (40 dB threshold elevation and clear histological damage) while the amplitude of wave 1 of the VsEP was not affected. It seems therefore that even though intense noise clearly affects the cochlea and may have a "masking" effect on the vestibular end organs, the intensities used in this study (113 dB SPL) are not able to produce a long-term noise induced vestibular disorder in the initially normal ear. These differences between the response of the cochlear and vestibular end organs to noise may be due to dissimilarities in their acoustic impedances and/or their electrical resting potential.


Phaeoviruses discovered in kelp (Laminariales).
Phaeoviruses are latent double-stranded DNA viruses that insert their genomes into those of their brown algal (Phaeophyceae) hosts. So far these viruses are known only from members of the Ectocarpales, which are small and short-lived macroalgae. Here we report molecular and morphological evidence for a new Phaeovirus cluster, referred to as sub-group C, infecting kelps (Laminariales) of the genera Laminaria and Saccharina, which are ecologically and commercially important seaweeds. Epifluorescence and TEM observations indicate that the Laminaria digitata Virus (LdigV), the type species of sub-group C, targets the host nucleus for its genome replication, followed by gradual degradation of the chloroplast and assembly of virions in the cytoplasm of both vegetative and reproductive cells. This study is the first to describe phaeoviruses in kelp. In the field, these viruses infected two thirds of their host populations; however, their biological impact remains unknown.


Nicotinic neuronal acetylcholine receptor alpha-3 subunit transcription in normal and myasthenic thymus.
Thymic transcription of the alpha-3 subunit of the AChR was studied through sequencing and PCR analysis of thymic cDNA clones, Northern blotting, and ribonuclease protection assays. This analysis revealed at least three, 3' end sequence variants for the alpha-3 subunit as well as a variant that results from the alternative splicing of an antisense 122 bp Alu sequence between exons 5 and 6 of the normal transcript. The spliced Alu sequence not only shifts the exon 6 reading frame but also carries an in-frame stop codon. If translated, this variant transcript would produce a truncated peptide lacking the fourth transmembrane domain of the subunit and carrying a carboxy terminus dodecapeptide not found in any other known AChR subunit sequence. The putative variant subunit may lack biological activity and should differ antigenically from its normal counterpart. In comparing the normal, the MG hypertrophic, and the MG thymoma for transcription of the alpha-3 subunit and its 122 bp variant, it was found that there were no qualitative or quantitative changes in alpha-3 transcript expression in the MG hypertrophic thymi. Thymomas, however, showed an overall decrease in alpha-3 transcription and a comparative increase in beta-amyloid precursor transcription. The decrease in the levels of alpha-3 transcription in thymomas may be related to the proliferation of thymic epithelial cells.


Expression of human blood group antigens A and B in kidney and lung of some vertebrates.
Human blood group antigens (BGA) are genetically determined glycoproteins found in many cells and tissues of different mammals. Their major biological functions are still undefined. There are few investigations analysing the evolutionary aspect of BGA tissue distribution. The present study is aimed at examining the expression of human A and B antigens in the kidney and lung of some free-living vertebrates. The biotin-streptavidin-peroxidase immunostaining system was applied on kidney and lung paraffin sections derived from free-living representatives of five different vertebrate classes. Excluding the possibility of any non-specific staining by the application of inhibition tests, A and B antigens were demonstrated most constantly in epithelial cells of renal and respiratory tubules. They were also detected in chondrocytes of fish gills, in some muscular and endothelial cells. Single erythrocytes showed a positive cytoplasmic staining only in some higher vertebrates. Human BGA seem to be conserved carbohydrate structures with biological functions probably related to cell integrity and differentiation.


Serotonin-induced bistability of turtle motoneurones caused by a nifedipine-sensitive calcium plateau potential.
1. The effect of serotonin on the firing properties of motoneurones was studied in transverse sections of the adult turtle spinal cord in vitro with intracellular recording techniques. 2. In normal medium, turtle motoneurones adapt from an initial high frequency to a low steady firing during a depolarizing current pulse. In the presence of serotonin (4-100 microM) motoneurones responded with accelerated firing and a frequency jump during a depolarizing current pulse followed by an after-depolarization outlasting the stimulus. From a depolarized holding potential motoneuronal activity was shifted between two stable states by brief depolarizing and hyperpolarizing current pulses. As an expression of this bistable firing behaviour, the frequency-current relation in response to a triangular current injection was counter-clockwise in serotonin while clockwise in normal medium. 3. The delay to onset of the frequency jump was shortened as the amplitude of the activation pulse was increased. From a positive holding potential the after-depolarization exceeded spike threshold and its duration increased with an increase in steady bias current. The effect of serotonin on turtle motoneurones could be blocked by methysergide (10 microM). 4. When action potentials were depressed by tetrodotoxin, a voltage-dependent, non-inactivating plateau potential, intrinsic to the motoneurone, was revealed. Activation of this voltage plateau provides the motoneurones with two stable states of firing. The apparent input resistance was 2-4-fold lower during the plateau than at rest. 5. The serotonin-induced plateau potential was Ca2+-dependent and was blocked when Ca2+ was replaced by either Co2+ (3 mM) or Mn2+ (3 mM). 6. The Ca2+ plateau was blocked by nifedipine (1-15 microM). 7. Serotonin reduced the slow after-hyperpolarization following action potentials. The change in balance between inward and outward currents seems to be sufficient to reveal the plateau response. 8. Although a small plateau response was induced by Bay K 8644 (1-15 microM), this L-channel agonist could not reproduce the pronounced effect of serotonin. 9. It is concluded that serotonin induces a Ca2+-dependent and nifedipine-sensitive plateau potential in turtle motoneurones primarily by reducing a K+-current responsible for the slow after-hyperpolarization.


Proteomic profiling of advanced melanoma patients to predict therapeutic response to anti-PD-1 therapy.
Despite high clinical need, there are no biomarkers that accurately predict the response of patients with metastatic melanoma to anti-PD-1 therapy. In this multicenter study, we applied protein depletion and enrichment methods prior to various proteomic techniques to analyze a serum discovery cohort (n=56) and three independent serum validation cohorts (n=80; n=12; n=17). Further validation analyses by literature and survival analysis followed. We identified several significantly regulated proteins as well as biological processes such as neutrophil degranulation, cell-substrate adhesion, and extracellular matrix organization. Analysis of the three independent serum validation cohorts confirmed the significant differences between responders (R) and non-responders (NR) observed in the initial discovery cohort. In addition, literature-based validation highlighted 30 markers overlapping with previously published signatures. Survival analysis using the TCGA database showed that overexpression of 17 of the markers we identified correlated with lower overall survival in melanoma patients. Ultimately, this multilayered serum analysis led to a potential marker signature with 10 key markers significantly altered in at least two independent serum cohorts: CRP, LYVE1, SAA2, C1RL, CFHR3, LBP, LDHB, S100A8, S100A9, and SAA1, which will serve as the basis for further investigation. In addition to patient serum, we analyzed primary melanoma tumor cells from NR and found a potential marker signature with 4 key markers: LAMC1, PXDN, SERPINE1, and VCAN.


Mice deficient in nuclear factor of activated T-cell transcription factor c2 mount increased Th2 responses after infection with Nippostrongylus brasiliensis and decreased Th1 responses after mycobacterial infection.
Infection of nuclear factor of activated T-cell transcription factor c2 (NFATc2)-deficient mice with the helminth Nippostrongylus brasiliensis led to a distinct increase in interleukin-4 (IL-4) and IL-5 protein synthesis by lymph node and spleen cells and to elevated serum immunoglobulin E (IgE) levels in comparison to those seen with infected control mice. While IL-4, IL-5, and IL-13 mRNA expression was also enhanced in lymph node cells from the lungs of infected NFATc2(-/-) mice, the number of T cells secreting Th2-type lymphokines remained the same in mice infected with N. brasiliensis. In contrast, lymphocytes from NFATc2-deficient mice infected with Mycobacterium bovis BCG secreted less gamma interferon than lymphocytes from infected control mice. These findings indicate that NFATc2 is an activator of Th1 responses and a suppressor of Th2 responses in vivo.


Sugammadex: A Limited But Important Role in Emergency Medicine.
Sugammadex reverses neuromuscular blockade by the steroidal nondepolarizing neuromuscular blocking agents rocuronium and vecuronium. In 2015, it was approved in the United States by the Food and Drug Administration for adult use. However, there are ongoing clinical trials investigating its use in the pediatric population. Before approval in adult use in the United States, several adverse effects were noted to occur in patients receiving sugammadex in clinical trials including prolonged QT interval, bradycardia, hypersensitivity reactions, and prolongation of coagulation parameters. Additional investigations further elucidated the risks of these adverse events. Sugammadex is approved for use in children older than 2 years in other countries in Europe and Asia. Investigations suggest that the efficacy, safety, and pharmacokinetic profile is similar in children when compared with adults. Published pediatric data favor the use of sugammadex in children older than 2 years, but there are some data in young children younger than 2 years. Case reports discuss the use of sugammadex in pediatric patients with neuromuscular diseases. Although sugammadex is typically used in the operating room for reversing neuromuscular blockade for surgical procedures, there is a small but important role for sugammadex use in the emergency department. In cases where rapid neurological examination is required after neuromuscular blockage with rocuronium or vecuronium, sugammadex can assist in facilitating a timely comprehensive neurological examination where pharmacologic or surgical management may depend on examination findings such as in the case of cerebral vascular accident, status epilepticus, or traumatic brain injury. Some clinicians have advocated for the use of sugammadex in the cannot intubate, cannot ventilate scenario. However, caution should be exercised in this situation as reversal of paralysis can take up to 22 minutes to occur.


Green's function modeling of response of two-dimensional materials to point probes for scanning probe microscopy.
A Green's function (GF) method is developed for interpreting scanning probe microscopy (SPM) measurements on new two-dimensional (2D) materials. GFs for the Laplace/Poisson equations are calculated by using a virtual source method for two separate cases of a finite material containing a rectangular defect and a hexagonal defect. The prescribed boundary values are reproduced almost exactly by the calculated GFs. It is suggested that the GF is not just a mathematical artefact but a basic physical characteristic of material systems, which can be measured directly by SPM for 2D solids. This should make SPM an even more powerful technique for characterization of 2D materials.


Organophosphate-induced acetylcholinesterase inhibition and embryonic retinal cell necrosis in vivo in the teleost (Oryzias latipes).
Recent monitoring of the Sacramento-San Joaquin River system (CA) indicates that levels of the organophosphate pesticide, diazinon, exceed National Academy of Science guidelines and these levels result in toxicity in USEPA acute toxicity tests with Ceriodaphnia dubia. Since organophosphates (OPs) inhibit acetylcholinesterase (AChE), the present study examined the effects of diazinon on the embryonic nervous system of a model teleost, medaka, Oryzias latipes. Preliminary histological screens revealed limited retinal cell necrosis in control embryos with apparent increased necrosis in diazinon-exposed embryos. Subsequently, embryos were exposed to 1.8 x 10(-5), 4.4 x 10(-5), or to 8.8 x 10(-5) M diazinon and replicates were frozen for biochemical analysis or were fixed for histopathological analysis at days 3, 5, and 7 of development. Diazinon exposure significantly inhibited AChE activity within whole embryos and in homogenates of retinas from treated animals. Histological examination of embryos indicated that as the retina underwent differentiation into distinct cell layers, between days 5 and 7, small foci of necrotic cells became apparent within the inner nuclear layer and isolated individual pyknotic cells were observed in the ganglion layer. Quantification of foci of necrotic cells revealed that 8.8 x 10(-5) M diazinon increased number and area of these lesions. Enzyme histochemistry localized AChE activity to regions equivalent to sites of necrosis. Separate exposures of embryos to the OP, diisopropylphosphorofluoridate, produced large foci of necrotic cells at sites equivalent to those seen following diazinon exposure.


Distinct timescales of RNA regulators enable the construction of a genetic pulse generator.
To build complex genetic networks with predictable behaviors, synthetic biologists use libraries of modular parts that can be characterized in isolation and assembled together to create programmable higher-order functions. Characterization experiments and computational models for gene regulatory parts operating in isolation are routinely used to predict the dynamics of interconnected parts and guide the construction of new synthetic devices. Here, we individually characterize two modes of RNA-based transcriptional regulation, using small transcription activating RNAs (STARs) and clustered regularly interspaced short palindromic repeats interference (CRISPRi), and show how their distinct regulatory timescales can be used to engineer a composed feedforward loop that creates a pulse of gene expression. We use a cell-free transcription-translation system (TXTL) to rapidly characterize the system, and we apply Bayesian inference to extract kinetic parameters for an ordinary differential equation-based mechanistic model. We then demonstrate in simulation and verify with TXTL experiments that the simultaneous regulation of a single gene target with STARs and CRISPRi leads to a pulse of gene expression. Our results suggest the modularity of the two regulators in an integrated genetic circuit, and we anticipate that construction and modeling frameworks that can leverage this modularity will become increasingly important as synthetic circuits increase in complexity.


An integrated and multi-functional droplet-based microfluidic platform for digital DNA amplification.
Digital DNA amplification is a powerful method for detecting and quantifying rare nucleic acids. In this study, we developed a multi-functional droplet-based platform that integrates the traditional digital DNA amplification workflow into a one-step device. This platform enables efficient droplet generation, transition, and signal detection within a 5-min timeframe, distributing the sample into a uniform array of 4 × 104 droplets (variation <2%) within a chamber. Subsequent in-situ DNA amplification, fluorescence detection, and signal analysis were carried out. To assess the platform's performance, we quantitatively detected the human epidermal growth factor receptor (EGFR) mutation and human papillomavirus (HPV) mutation using digital polymerase chain reaction (dPCR) and digital loop-mediated isothermal amplification (dLAMP), respectively. The fluorescence results exhibited a positive, linear, and statistically significant correlation with target DNA concentrations ranging from 101 to 105 copies/μL, demonstrating the capability and feasibility of the integrated device for dPCR and dLAMP. This platform offers high-throughput droplet generation, eliminates droplet fusion and transition, is user-friendly, reduces costs compared to current methods, and holds potential for thermocycling and isothermal nucleic acid quantification with high sensitivity and accuracy.


Covariance, feedback, and discounting in ratio schedules.
Action-outcome relations in the everyday world are often looser than the relations programmed in the laboratory. Reinforcement theory addressed this looseness by theorizing about "delay of reinforcement." Response-reinforcer interval affects response rate and choice, but need not be thought of as delay. Experiments on temporal discounting are often said to reveal effects of delay also, but the intervals chosen differ from delays, because an outcome after a week, month, or year occurs after life has continued and many events have intervened. They are better called lag intervals. The present experiment studied the effects of varying lag interval on pigeons' responding on ratio-like schedules of food. In a continuous procedure, a lag interval separated pecking that earned food from delivery of the food earned. In one series of conditions, the relation was fixed equivalent to variable-ratio (VR) 40 while the lag interval varied. In a second series, the equivalent VR varied while the lag interval was fixed. The results accorded with matching theory, but with a factor for temporal discounting or loose feedback. The results suggest that both temporal discounting and probability discounting may be characterized within matching theory, but that temporal discounting requires a separate treatment from probability discounting.


Topiroxostat versus allopurinol in patients with chronic heart failure complicated by hyperuricemia: A prospective, randomized, open-label, blinded-end-point clinical trial.
The benefits of xanthine oxidase inhibitors to chronic heart failure (CHF) patients is controversial. We investigated the beneficial effects of a novel xanthine oxidoreductase inhibitor, topiroxostat, in patients with CHF and hyperuricemia (HU), in comparison to allopurinol. The prospective, randomized open-label, blinded-end-point study was performed in 141 patients with CHF and HU at 4 centers. Patients were randomly assigned to either topiroxostat or allopurinol group to achieve target uric acid level ≤6.0 mg/dL. According to the protocol, 140 patients were followed up for 24 weeks. Percent change in ln (N-terminal-proB-type natriuretic peptide) at week 24 (primary endpoint) was comparable between topiroxostat and allopurinol groups (1.6±8.2 versus -0.4±8.0%; P = 0.17). In the limited number of patients with heart failure with reduced ejection fraction (HFrEF) (left ventricle ejection fraction <45%), ratio of peak early diastolic flow velocity at mitral valve leaflet to early diastolic mitral annular motion velocity (E/e') decreased in topiroxostat group, but not in allopurinol group. Urinary 8-hydroxy-2'-deoxyguanosine and L-type fatty acid-binding protein levels increased and osmolality decreased significantly in allopurinol group, while these changes were less or absent in topiroxostat group. In allopurinol group HFrEF patients, additional to the increases in these urinary marker levels, urinary creatinine levels decreased, with no change in clearance, but not in topiroxostat group. Compared with allopurinol, topiroxostat did not show great benefits in patients with CHF and HU. However, topiroxostat might have potential advantages of reducing left ventricular end-diastolic pressure, not worsening oxidative stress in proximal renal tubule, and renoprotection over allopurinol in HFrEF patients.


Symptom control, functioning, and hospitalization status in French patients changed from oral atypical antipsychotics to risperidone long-acting injectable.
Objective. To investigate efficacy and tolerability of risperidone long-acting injectable (RLAI) in French patients with schizophrenia or other psychotic disorders, who were all switching from previous treatment with oral atypical antipsychotics. The impact of treatment with RLAI on the hospitalization status of these patients was also examined. Methods. Clinically stable patients requiring a treatment change received 25 mg RLAI (increased to 37.5 or 50 mg if required) every 2 weeks for 6 months. Results. Of 130 patients (68.5% male, mean age 36.2 years), most (83.8%) had DSM-IV schizophrenia (mainly paranoid). Previous treatments were risperidone (80.8%), olanzapine (10.0%) and amisulpride (10.0%). Out of 66 patients hospitalized at baseline, 51 were outpatients at endpoint. Mean total PANSS, CGI-S and GAF scores were significantly reduced from baseline to treatment endpoint (p<0.001). Of those patients reported as moderate to severely ill at the beginning of the trial (81.3%), fewer had the same classification at endpoint (50.8%). Mean scores for total ESRS and Parkinsonism subscales were significantly reduced after only 1 month of treatment (p<0.001). Conclusion. Treatment with RLAI significantly improved disease symptoms, functioning, hospitalization status, and reduced movement disorders, in psychotic patients considered clinically stable on oral atypical antipsychotics.


Comparing newer and older analgesics and delivery systems.
The newer opioids, rather than replacing older agents, add to the options for treating pain. The particular pharmacodynamic and pharmacokinetic features of these drugs determine the best choice for a given patient.


Acculturation and Health Insurance of Mexicans in the USA.
We study how the health insurance coverage of Mexican immigrants changes with time in the U.S. Cross sectional estimates indicate that time since arrival is negatively correlated with the probability of being uninsured for both male and female Mexican immigrants, and about a third of the decline could be attributed to civic and labor market incorporation of Mexican immigrants. However, much of the relationship between time in the U.S. and health insurance coverage, after adjusting for demographic and labor market factors, is due to failure to control for age at arrival and period of arrival. Estimates from longitudinal analyses suggest that there is no systematic relationship between time in the U.S. and health insurance of Mexican immigrants, although imprecision in the fixed effects estimates makes it difficult to draw firm conclusions.


Multiple sclerosis presenting as ataxic hemiparesis.
Multiple sclerosis (MS) can present with many clinical pictures, but only rarely as a lacunar syndrome. The following case presentation is of multiple sclerosis with initial presentation as ataxic hemiparesis (AH), indistinguishable from the vascular syndrome. This case serves to illustrate that even classic lacunar syndromes can actually be the initial manifestation of multiple sclerosis and that other etiologies besides ischemic stroke need to remain in the differential diagnosis until a definitive has been concluded.


Synergistic activation of human platelets by lysophosphatidic acid and adrenaline.
Platelet reactivity is regulated by various important bioactive and physiologic substances. The objective of this study was to characterize lysophosphatidic acid (LPA)-triggered responses in human platelets. In addition, the effect of LPA was compared with that of other activators and possible synergistic interactions were evaluated. LPA-triggered cytosolic Ca(2+) responses were measured using fura-2-loaded platelets in a spectrofluorometer. Furthermore, platelet aggregation and secretion were analyzed in a lumi-aggregometer and protein tyrosine phosphorylation was detected with the Western blot technique. LPA dose-dependently increased cytosolic Ca(2+) concentration ([Ca(2+)](i)) in platelets. This response involved both influx of extracellular Ca(2+) and release of Ca(2+) from intracellular stores. However, in comparison with other platelet agonists, i.e. thrombin and adenosine 5'-diphosphate (ADP), LPA was a very weak Ca(2+)-elevating agent. Furthermore, we observed that the LPA-induced rise in [Ca(2+)](i) was markedly suppressed by cyclic nucleotide-elevating agents. In functional studies, LPA failed to stimulate platelet aggregation and secretion. However, in combination with adrenaline, another weak platelet agonist, LPA could induce an irreversible and complete aggregatory response. There was an individual variation in aggregatory response and tyrosine phosphorylation when LPA and adrenaline were combined. These agents induced a powerful response on platelets from some individuals, but had a weak or no effect on others. The present study shows, for the first time, that isolated platelets from some healthy blood donors respond synergistically to a combination of LPA and adrenaline. Platelet activation is a key step in distinguishing normal hemostasis from pathologic hemostasis. Increased knowledge about this mechanism might help to predict individual responses and provide new insights into molecular mechanisms responsible for pathologic thrombosis.


Polarized Two-Photon Absorption and Heterogeneous Fluorescence Dynamics in NAD(P)H.
Two-photon absorption (2PA) finds widespread application in biological systems, which frequently exhibit heterogeneous fluorescence decay dynamics corresponding to multiple species or environments. By combining polarized 2PA with time-resolved fluorescence intensity and anisotropy decay measurements, we show how the two-photon transition tensors for the components of a heterogeneous population can be separately determined, allowing structural differences between the two fluorescent states of the redox cofactor NAD(P)H to be identified. The results support the view that the two states correspond to alternate configurations of the nicotinamide ring, rather than folded and extended conformations of the entire molecule.


Neutralization of LPS or blockage of TLR4 signaling prevents stress-triggered fetal loss in murine pregnancy.
Maternal stress can cause loss of both histocompatible (syngeneic) and histoincompatible (semiallogeneic) embryos in pregnant mice. Stress increases abortogenic Th1 cytokines and reduces levels of anti-abortogenic Th2 cytokines, progesterone levels, and T regulatory cell activity. While physiological levels of interferon-γ promote vascular remodeling at the feto-maternal interface, an overshooting Th1 cytokine response requires a Toll-like receptor (TLR)-mediated "danger signal" such as lipopolysaccharide (LPS). Interestingly, stress can enhance permeability of mucosal membranes to entry of bacterial products and promote transmucosal migration of commensal bacteria. We hypothesized that bacterial component such as LPS may provide the danger signal through which stress triggers maternal immune activation, subsequently resulting in fetal rejection. Blocking the TLR4 receptor for LPS or neutralization of LPS using bactericidal permeability increasing protein abrogate fetal loss due to sonic stress challenge in DBA/2J-mated CBA/J mice. These treatments prevented stress-triggered immune responses in the decidua, upregulated Treg cells, and reduced the frequency of mature dendritic cells in uterine-draining lymph nodes but not in the uterus. Interestingly, anti-TLR4 treatment only partly ameliorated stress-induced endocrine responses, such as increased hypothalamic corticotropin releasing hormone and vasopressin mRNA expression but not decrease of serum progesterone. Galectin-1 knock-out mice were more susceptible to stress-triggered complete implantation failure rather than fetal loss, which was also abolished by LPS neutralization. Insights provided in this paper shed new light on the mechanisms by which stress affects pregnancy outcome and introduce microbial-derived LPS as a mediator within the cascade of stress-triggered immune and endocrine events during pregnancy.


Alexithymia and Psychopathology in Patients Suffering From Inflammatory Bowel Disease: Arising Differences and Correlations to Tailoring Therapeutic Strategies.
Comorbidity with anxiety or depression is common in patients with Inflammatory Bowel Disease (IBD) as Crohn Disease (CD) and Ulcerative Colitis (UC). Data suggest that the cognitive construct of alexithymia has high prevalence in people suffering from anxiety and mood disorders and even in people with IBD. Most studies have investigated mainly anxiety and depression, considering IBD population as a homogeneous group of patients. Little evidence shows the impact of alexithymia on the course of IBD. We evaluated a broad spectrum of psychopathological symptoms and alexithymia levels in a group of outpatients affected by IBD in clinical remission, comparing CD and UC and investigating the relationship with clinical and socio-demographic variables. One hundred and seventy IBD outpatients were screened by using the Hospital Anxiety Depression Scale (HADS), the Self-report Symptom Inventory-90-Revised (SCL-90-R) and the Toronto Alexithymia Scale (TAS-20). A high prevalence of anxious and depressive symptoms (42.35 and 25.8% respectively) together with alexithymia (31.76%) was confirmed. CD patients experienced high levels of depression (HADS Depression 35.2% p = 0.034; SCL-90-R mean 1.39 p < 0.001), somatisation (SCL-90-R mean 1.04 p < 0.001), obsessive-compulsive symptoms (SCL-90-R mean 1.2 p < 0.001), and global severity (SCL-90-R mean 1.15 p < 0.001). There is no statistical difference in the prevalence of alexithymia in both subpopulations. The levels of alexithymia are correlated to the levels of anxiety (HADS Anxiety rs = 0.516 p < 0.001), depression (HADS Depression rs = 0.556 p < 0.001; SCL-90-R rs = 0.274 p = 0.001), somatisation (SCL-90-R rs = 0.229 p = 0.005), obsessive-compulsive symptoms (SCL-90-R rs = 0.362 p < 0.001), and global severity (SCL-90-R rs = 0.265 p = 0.001). Furthermore, alexithymia is associated with a delay of diagnosis of IBD, poly-therapies and greater IBD extension. Older age, female gender, greater IBD extension, surgery, and delay of diagnosis seem to be related to a high prevalence of psychopathological symptoms such as anxiety, depression, somatisation, and obsessive-compulsive symptoms. Psychopathological symptoms and high levels of alexithymia are frequent in IBD patients and seem to be related to a high risk of poor clinical outcome. CD patients could be considered at higher risk of mental comorbidity. A more comprehensive psychiatric assessment, including alexithymia, and an integrated treatment of underlying conditions, must be taken into account in order to improve the global prognosis of the disease.


Effect of hyperglycemia on gastric acid secretion during the gastric phase of digestion.
We have examined the effect of an acute stable hyperglycemia on gastric acid secretion during the gastric phase of digestion. Gastric acid output was measured with a recovery marker (phenol red) under basal conditions and after repeated intragastric instillation of a liquid meal in seven healthy subjects on two separate occasions: during normoglycemia (serum glucose, 15 mM). Premeal gastric acid output was significantly (P < 0.05) reduced during hyperglycemia compared with during normoglycemia (2.6 +/- 1.0 vs. 5.8 +/- 1.8 mmol/h). Intragastric meal-stimulated incremental acid output during hyperglycemia was significantly (P < 0.05) reduced compared with during normoglycemia (19 +/- 4 vs. 38 +/- 9 mmol/120 min). Meal-stimulated gastrin release during hyperglycemia was significantly (P < 0.05) reduced compared with that during normoglycemia (4.9 +/- 1.3 vs. 6.6 +/- 1.6 micrograms.1(-1).120 min-1). The intragastric meal induced significant (P < 0.05) increases in pancreatic polypeptide concentrations only during normoglycemia. During hyperglycemia, recovery rates of gastric contents were significantly (P < 0.05) increased compared with during normoglycemia, both before (81 +/- 4 vs. 71 +/- 6%) and after (72 +/- 4 vs. 57 +/- 4%) meal ingestion, pointing to delayed gastric emptying of liquids during hyperglycemia. In conclusion, 1) gastric acid secretion under unstimulated conditions and during the gastric phase of digestion is reduced during hyperglycemia; 2) meal-stimulated gastrin release is significantly reduced during hyperglycemia; 3) the reduction in meal-stimulated acid output is correlated with the reduction in gastrin releases; and 4) pancreatic polypeptide secretion is significantly reduced during hyperglycemia, pointing to impaired vagal cholinergic tone.


Cytostatic activity of peptide extracts of medicinal plants on transformed A549, H1299, and HeLa Cells.
Biological activity of peptide extracts of medicinal plants was studied on transformed non-small-cell lung carcinoma A549 cells, lung cancer H1299 cells, and cervical cancer HeLa cells at various cell densities. Cell survival and proliferation were evaluated 72 h after treatment with extracts in concentrations of 0.05, 0.25, and 0.5 microg/microl. The cytostatic effect was produced by peptide extracts of Camelia sinesis Kuntze, Inonotus obliquus, and a mixture Inula helenium L., Chelidonium majus L., Equisetum arvense L., and Inonotus obliquus. Peptide extracts of Hypericum perforatum L. and Laurus nobilis L. in the same concentrations had no effects on proliferative activity and growth of tumor cells.


Transcriptome profiling of Variovorax paradoxus EPS under different growth conditions reveals regulatory and structural novelty in biofilm formation.
We used transcriptome analysis by paired-end strand-specific RNA-seq to evaluate the specific changes in gene expression associated with the transition to static biofilm growth in the rhizosphere plant growth-promoting bacterium Variovorax paradoxus EPS. Triplicate biological samples of exponential growth, stationary phase and static biofilm samples were examined. DESeq2 and Rockhopper were used to identify robust and widespread shifts in gene expression specific to each growth phase. We identified 1711 protein-coding genes (28%) using DESeq2 that had altered expression greater than twofold specifically in biofilms compared to exponential growth. Fewer genes were specifically differentially expressed in stationary-phase culture (757, 12%). A small set of genes (103/6020) were differentially expressed in opposing fashions in biofilm and stationary phase, indicating potentially substantial shifts in phenotype. Gene-ontology analysis showed that the only class of genes specifically upregulated in biofilms was associated with nutrient transport, highlighting the importance of nutrient uptake in the biofilm. The biofilm-specific genes did not overlap substantially with the loci identified by mutagenesis studies, although some were present in both sets. The most highly upregulated biofilm-specific gene is predicted to be a part of the RNA degradosome, which indicates that RNA stability is used to regulate the biofilm phenotype. Two small putative proteins, Varpa_0407 and Varpa_3832, are highly expressed specifically in biofilms and are predicted to be secreted DNA-binding proteins, which may stabilize extracellular DNA as a component of the biofilm matrix. An flp/tad type-IV pilus locus (Varpa_5148-60) is strongly downregulated specifically in biofilms, in contrast with results from other systems for these pili. Mutagenesis confirms that this locus is important in surface motility rather than biofilm formation. These experimental results suggest that V. paradoxus EPS biofilms have substantial regulatory and structural novelty.


Comparison between the two-step and the three-step algorithms for the detection of toxigenic Clostridium difficile.
To evaluate usefulness of applying either the two-step algorithm (Ag-EIAs and CCNA) or the three-step algorithm (all three assays) for better confirmation of toxigenic Clostridium difficile. The antigen enzyme immunoassays (Ag-EIAs) can accurately identify the glutamate dehydrogenase antigen of toxigenic and nontoxigenic Clostridium difficile. Therefore, it is used in combination with a toxin-detecting assay [cell line culture neutralization assay (CCNA), or the enzyme immunoassays for toxins A and B (TOX-A/BII EIA)] to provide specific evidence of Clostridium difficile-associated diarrhoea. A total of 151 nonformed stool specimens were tested by Ag-EIAs, TOX-A/BII EIA, and CCNA. All tests were performed according to the manufacturer's instructions and the results of Ag-EIAs and TOX-A/BII EIA were read using a spectrophotometer at a wavelength of 450 nm. A total of 61 (40.7%), 38 (25.3%), and 52 (34.7%) specimens tested positive with Ag-EIA, TOX-A/BII EIA, and CCNA, respectively. Overall, the sensitivity, specificity, negative predictive value, and positive predictive value for Ag-EIA were 94%, 87%, 96.6%, and 80.3%, respectively. Whereas for TOX-A/BII EIA, the sensitivity, specificity, negative predictive value, and positive predictive value were 73.1%, 100%, 87.5%, and 100%, respectively. With the two-step algorithm, all 61 Ag-EIAs-positive cases required 2 days for confirmation. With the three-step algorithm, 37 (60.7%) cases were reported immediately, and the remaining 24 (39.3%) required further testing by CCNA. By applying the two-step algorithm, the workload and cost could be reduced by 28.2% compared with the three-step algorithm. The two-step algorithm is the most practical for accurately detecting toxigenic Clostridium difficile, but it is time-consuming.


Flash visual evoked potentials in diurnal birds of prey.
The objective of this pilot study was to evaluate the feasibility of Flash Visual Evoked Potentials (FVEPs) testing in birds of prey in a clinical setting and to describe the protocol and the baseline data for normal vision in this species. FVEP recordings were obtained from 6 normal adult birds of prey: n. 2 Harris's Hawks (Parabuteo unicinctus), n. 1 Lanner Falcon (Falco biarmicus), n. 2 Gyrfalcons (Falco rusticolus) and n. 1 Saker Falcon (Falco cherrug). Before carrying out VEP tests, all animals underwent neurologic and ophthalmic routine examination. Waveforms were analysed to identify reproducible peaks from random variation of baseline. At least three positive and negative peaks were highlighted in all tracks with elevated repeatability. Measurements consisted of the absolute and relative latencies of these peaks (P1, N1, P2, N2, P3, and N3) and their peak-to-peak amplitudes. Both the peak latency and wave morphology achieved from normal animals were similar to those obtained previously in other animal species. This test can be easily and safely performed in a clinical setting in birds of prey and could be useful for an objective assessment of visual function.


Risk of aortic valve replacement in patients with aortic stenosis and chronic obstructive pulmonary disease.
Patients with aortic stenosis (AS) and chronic obstructive pulmonary disease (COPD) have been considered at high risk for aortic valve replacement (AVR), which results in some patients being denied this life-saving operation. Hence, the study aim was to assess the operative, 30-day, and long-term mortality in individuals with COPD undergoing AVR for AS in the modern surgical era. This retrospective cohort of patients had documented COPD (FEV1/FVC < 70%), and underwent isolated AVR for severe AS between 1993 and 2007 at the Mayo Clinic in Rochester, MN. Of the 68 patients who met the study criteria, 27 had mild/moderate COPD (FEV1 > 50%), 35 had severe COPD (FEV1 30-50%), and six had very severe COPD (FEV1 < 30%). The overall operative and 30-day mortality was 4.8%. More severe COPD was associated with a longer stay in the intensive care unit (42 h for mild/moderate versus 115 h for severe/very severe: p = 0.02), but did not influence the operative or 30-day mortalities. Female gender was associated with an increased length of hospital stay. Long-term mortality was significantly higher in patients with a history of cerebrovascular disease (HR 4.3, p < 0.001), NYHA class III or IV heart failure (class III HR 2.79, p = 0.05; class IV HR 3.97, p = 0.03), and increased age (HR 1.06, p = 0.003). The severity of COPD was an independent risk factor for long-term mortality. Patients with severe AS and COPD are at an acceptable risk for AVR (30-day mortality < 5%). The severity of COPD is not associated with an increased in-hospital or 30-day mortality, but does influence long-term mortality.


Changes in transfer ribonucleic acids of Bacillus subtilis during different growth phases.
The transfer ribonucleic acids (tRNAs) of B. subtilis at different growth phases are examined for changes in the composition and the methylation of minor constituents. The composition of the tRNAs indicates about equal amounts of adenosine and uridine, and of guanosine and cytidine. About 3-4 residues are present as modified bases in the average tRNA molecule. The net composition of tRNAs appears to remain unaltered during different growth phases. In vitro methylation of tRNAs indicates lack of methyl groups in both exponentially growing cells and spores. In vivo methylation studies show tRNA methylation occurs during the stationary phase in the absence of net tRNA synthesis. Thus, both in vitro and in vivo methylation indicate that the tRNAs in exponentially growing cells do not contain their full complement of modified bases. More complete modification is noted in tRNAs from stationary cells or spores. Hence, tRNA mofifications in general are preserved with fidelity even in the dormant spore but the possibility is left open that specific modifications of selected isoacceptors of tRNAs may occur.


Annual trend of neonatal mortality and its underlying causes: population-based study - São Paulo State, Brazil, 2004-2013.
Population-based studies analyzing neonatal deaths in middle-income countries may contribute to design interventions to achieve the Sustainable Development Goals, established by United Nations. This study goal is to analyze the annual trend of neonatal mortality in São Paulo State, Brazil, over a 10-year period and its underlying causes and to identify maternal and neonatal characteristics at birth associated with neonatal mortality. A population-based study of births and deaths from 0 to 27 days between 2004 and 2013 in São Paulo State, Brazil, was performed. The annual trend of neonatal mortality rate according to gestational age was analyzed by Poisson or by Negative Binomial Regression models. Basic causes of neonatal death were classified according to ICD-10. Association of maternal demographic variables (block 1), prenatal and delivery care variables (block 2), and neonatal characteristics at birth (block 3) with neonatal mortality was evaluated by Poisson regression analysis adjusted by year of birth. Among 6,056,883 live births in São Paulo State during the study period, 48,309 died from 0 to 27 days (neonatal mortality rate: 8.0/1,000 live births). For the whole group and for infants with gestational age 22-27, 28-31, 32-36, 37-41 and ≥ 42 weeks, reduction of neonatal mortality rate was, respectively, 18 %, 15 %, 38 %, 53 %, 31 %, and 58 %. Median time until 50 % of deaths occurred was 3 days. Main basic causes of death were respiratory disorders (25 %), malformations (20 %), infections (17 %), and perinatal asphyxia (7 %). Variables independently associated with neonatal deaths were maternal schooling, prenatal care, parity, newborn sex, 1st minute Apgar, and malformations. Cesarean delivery, compared to vaginal, was protective against neonatal mortality for infants at 22-31 weeks, but it was a risk factor for those with 32-41 weeks. Despite the significant decrease in neonatal mortality rate over the 10-year period in São Paulo State, improved access to qualified health care is needed in order to avoid preventable neonatal deaths and increase survival of infants that need more complex levels of assistance.


Opioid-Free Analgesia Provides Pain Control Following Thumb Carpometacarpal Joint Arthroplasty.
We hypothesized that an opioid-free (OF), multimodal pain management pathway for thumb carpometacarpal (CMC) joint arthroplasty would not have inferior pain control compared with that of a standard opioid-containing (OC) pathway. This was a single-center, randomized controlled clinical trial of patients undergoing primary thumb CMC joint arthroplasty. Patients were randomly allocated to either a completely OF analgesic pathway or a standard OC analgesic pathway. Patients in both cohorts received a preoperative brachial plexus block utilizing 30 mL of 0.5% ropivacaine that was administered via ultrasound guidance. The OF group was given a combination of cryotherapy, anti-inflammatory medications, acetaminophen, and gabapentin. The OC group was only given cryotherapy and opioid-containing medication for analgesia. Patient-reported pain was assessed with use of a 0 to 10 numeric rating scale at 24 hours, 2 weeks, and 6 weeks postoperatively. We compared the demographics, opioid-related side effects, patient satisfaction, and Veterans RAND 12-Item Health Survey (VR-12) results between these 2 groups. At 24 hours postoperatively, pain scores in the OF group were statistically noninferior to, and lower than, those in the OC group (median, 2 versus 4; p = 0.008). Pain scores continued to differ significantly at 2 weeks postoperatively (median, 2 versus 4; p = 0.001) before becoming more similar at 6 weeks (p > 0.05). No difference was found between groups with respect to opioid-related side effects, patient satisfaction, or VR-12 results. A completely opioid-free perioperative protocol is effective for the treatment of pain following thumb CMC joint arthroplasty in properly selected patients. Therapeutic Level I . See Instructions for Authors for a complete description of levels of evidence.