Mechanism of action of CM-55, a synthetic analogue of the antilipogenic antibiotic cerulenin.
CM-55 is a synthetic analogue of the antibiotic cerulenin with the chemical structure of 2, 3-dodecenyl-4-oxo-dimethyl amide. This compound inhibited the growth of Saccharomyces cerevisiae ATCC 12341 and inhibited protein and lipid synthesis by 91 and 95%, respectively, at a concentration of 50 mug/ml (2.1 x 10(-4) M). The inhibition of protein synthesis was associated with the partial reduction of ribonucleic acid synthesis and leucine transport. The mechanism of inhibition of lipid synthesis was further investigated in a cell-free extract of the yeast. CM-55 inhibited the incorporation of [(14)C]acetyl Coenzyme A (CoA) into both fatty acid (FAF) and non-saponifiable fractions (NSF). However, it did not inhibit [(14)C]malonyl CoA incorporation into FAF and only slightly inhibited [(14)C]mevalonate incorporation into NSF. The activity of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) synthase was inhibited more strongly than the incorporation of [(14)C]3-hydroxy-3-methylglutaryl CoA into NSF; this could account for the CM-55 inhibition of [(14)C]acetyl CoA incorporation into NSF.


Multimarker approach to risk stratification for long-term mortality in patients on chronic hemodialysis.
We prospectively investigated the prognostic value of the combined use of cardiac troponin T (TnT), B-type natriuretic peptide (BNP), and high-sensitivity C-reactive protein (CRP) for long-term mortality in hemodialysis (HD) patients. METHODS AND RESULTS: Baseline measurements of TnT, BNP, and CRP were performed in 516 patients on chronic HD. Patients were followed up for 10 years. Using the Cox multivariate model with these 3 biomarkers as variables categorized into tertiles for mortality, a simplified score was obtained by underscoring individual biomarkers based on the adjusted hazard ratio (HR). The multimarker score was defined as the sum of these points. TnT, BNP, and CRP levels were individually independent predictors for mortality (P<0.05). Among low-risk (multimarker score <4), intermediate-risk (multimarker score 4-7), and high-risk (multimarker score ≥7) groups, 10-year survival rates were 83.3%, 54.3%, and 27.2% (P<0.0001), respectively. After adjusting for other confounders, the multimarker score had strong predictive power for mortality (HR: 4.26; P<0.0001 for high-risk vs. low-risk group). Furthermore, adding the multimarker score to a baseline model with established risk factors improved the C-index (P<0.01), net reclassification improvement (P<0.0001), and integrated discrimination improvement (P<0.0001) greater than that of any single biomarker or baseline model alone. The multimarker approach (ie, simultaneous assessment of TnT, BNP, and CRP, which individually independently predict prognosis) may improve the prediction of long-term mortality in HD patients.


High-speed helmeted head impacts in motorcycling: A computational study.
The motorcyclist is exposed to the risk of falling and impacting ground head-first at a wide range of travelling speeds - from a speed limit of less than 50 km/h on the urban road to the race circuit where speed can reach well above 200 km/h. However, motorcycle helmets today are tested at a single and much lower impact speed, i.e. 30 km/h. There is a knowledge gap in understanding the dynamics and head impact responses at high travelling speeds due to the limitation of existing laboratory rigs. This study used a finite element head model coupled with a motorcycle helmet model to simulate head-first falls at travelling speed (or tangential velocity at impact) from 0 to 216 km/h. The effect of different falling heights (1.6 m and 0.25 m) and coefficient of frictions (0.20 and 0.45) between the helmet outer shell and ground were also examined. The simulation results were analysed together with the analytical model to better comprehend rolling and/or sliding phenomena that are often observed in helmet oblique impacts. Three types of helmet-to-ground interactions are found when the helmet impacts ground from low to high tangential velocities: (1) helmet rolling without slipping; (2) a combination of sliding and rolling; and (3) continuous sliding. The tangential impulse transmitted to the head-helmet system, peak angular head kinematics and brain strain increase almost linearly with the tangential velocity when the helmet rolls but plateaus when the helmet slides. The critical tangential velocity at which the motion transit from the rolling regime to the sliding regime depends on both the falling height and friction coefficient. Typically, for a fall height of 1.63 m and a friction coefficient of 0.45, the rolling/sliding transition occurs at a tangential velocity of 10.8 m/s (38.9 km/h). Low sliding resistance in helmet design, i.e. by the means of a lower friction coefficient between the helmet outer shell and ground, has shown a higher reduction of brain tissue strain in the sliding regime than in the rolling regime. This study uncovers the underlying dynamics of rolling and sliding phenomena in high-speed oblique impacts, which largely affect head impact biomechanics. Besides, the study highlights the importance of testing helmets at speeds covering both the rolling and sliding regime since potential designs for improved head protection at high-speed impacts can be more distinguishable in the sliding regime than in the rolling regime.


Blood glucose control in critical care patients - a review of the literature.
The role of hyperglycaemia in critical illness, and its corresponding treatment, has been an area of controversy, fuelled by conflicting research findings. The aims of this study were to critically evaluate the literature and present an historical review of the sequence of published papers relating to blood glucose control in critical care. Their subsequent impact together with the implications for patient care is discussed. This article is based on a systematic review of papers relating to glycaemic control in critical care patients. The review was conducted using the MedLine, CINAHL and EMBASE databases using key search terms (details of the search terms can be found after the conclusion of the paper) for the period 1950-2006. The searches resulted in 4863 papers being screened for relevance to the historic progression of glycaemic management in critical care patients, by title and then abstract. Of these, 209 were accessed, and their reference lists were snowballed for further papers. Papers that were repeatedly quoted throughout the literature and were therefore considered important in the historical development of accepted critical care practice were finally subjected to rigorous appraisal. These totalled 91 papers and included 18 randomized controlled trials, an additional 28 research papers, 25 editorials and 20 reviews. This critical evaluation of published work indicates that the evidence for the benefit of this therapy may not be as compelling as previously indicated, and its widespread use may have been premature. From a nursing perspective, this demonstrates the importance of maintaining a questioning attitude to new therapies and reviewing best practice in the light of evolving evidence.


Measurement and explanation of socioeconomic inequality in catastrophic health care expenditure: evidence from the rural areas of Shaanxi Province.
Policy interventions have been taken to protect households from facing unpredictable economic changes that may cause catastrophe in China. This study aims to estimate the change of overall proportion of households incurring catastrophic health care expenditure (CHE) and its income-related inequality in the rural areas of Shaanxi Province from 2008 to 2013. The data were drawn from the National Household Health Service Surveys of Shaanxi Province conducted in the years 2008 and 2013. In total, 3,217 households in 2008 and 13,085 households in 2013 were selected for analysis. A "Capacity to pay" approach was used to measure the incidence of CHE. The concentration index was employed to measure the extent of income-related inequality in CHE. A decomposition method, based on a logit model, was used to decompose the concentration index into its determining components. From 2008 to 2013, the overall proportion of households incurring CHE dropped from 17.19 % to 15.83 %, while conversely, the inequality in facing CHE strongly increased. The majority of observed inequalities in CHE were explained by household economic status and household size in 2013. In addition, the absence of commercial health insurance and having elderly members were also important contributors to inequality in CHE. Even though we used a conservative method to measure CHE, the overall proportion of households incurring CHE in Shaanxi Province is still considerably high in both years. Furthermore, there exists a strong pro-rich inequality of CHE in rural areas of Shaanxi Province. Our study suggests that narrowing the gap of household economic status, improving the anti-risk capability of small scale households, establishing prepayment mechanisms in health insurance, strengthening the depth of reimbursement and subsidising vulnerable households in Shaanxi Province are helpful for both reducing the probability of incurring CHE and the pro-rich inequality in CHE.


The PCORnet Blood Pressure Control Laboratory: A Platform for Surveillance and Efficient Trials.
Uncontrolled blood pressure (BP) is a leading preventable cause of death that remains common in the US population despite the availability of effective medications. New technology and program innovation has high potential to improve BP but may be expensive and burdensome for patients, clinicians, health systems, and payers and may not produce desired results or reduce existing disparities in BP control. The PCORnet Blood Pressure Control Laboratory is a platform designed to enable national surveillance and facilitate quality improvement and comparative effectiveness research. The platform uses PCORnet, the National Patient-Centered Clinical Research Network, for engagement of health systems and collection of electronic health record data, and the Eureka Research Platform for eConsent and collection of patient-reported outcomes and mHealth data from wearable devices and smartphones. Three demonstration projects are underway: BP track will conduct national surveillance of BP control and related clinical processes by measuring theory-derived pragmatic BP control metrics using electronic health record data, with a focus on tracking disparities over time; BP MAP will conduct a cluster-randomized trial comparing effectiveness of 2 versions of a BP control quality improvement program; BP Home will conduct an individual patient-level randomized trial comparing effectiveness of smartphone-linked versus standard home BP monitoring. Thus far, BP Track has collected electronic health record data from over 826 000 eligible patients with hypertension who completed ≈3.1 million ambulatory visits. Preliminary results demonstrate substantial room for improvement in BP control (<140/90 mm Hg), which was 58% overall, and in the clinical processes relevant for BP control. For example, only 12% of patients with hypertension with a high BP measurement during an ambulatory visit received an order for a new antihypertensive medication. The PCORnet Blood Pressure Control Laboratory is designed to be a reusable platform for efficient surveillance and comparative effectiveness research; results from demonstration projects are forthcoming.


Transport of the natural sweetener stevioside and its aglycone steviol by human organic anion transporter (hOAT1; SLC22A6) and hOAT3 (SLC22A8).
The natural sweetening agent stevioside and its aglycone metabolite, steviol, have been shown to inhibit transepithelial transport of para-aminohippurate (PAH) in isolated rabbit renal proximal tubules by interfering with basolateral entry. The aim of the present study was to determine which of the cloned basolateral organic anion transporters were involved in the renal transport of stevioside and steviol. This question was addressed in Xenopus laevis oocytes expressing human organic anion transporter 1 (hOAT1), 3 (hOAT3), and winter flounder OAT (fOat1). The parent compound, stevioside, had no inhibitory effect on either PAH (hOAT1) or ES (estrone sulfate; hOAT3) uptake. In contrast, steviol showed significant, dose-dependent inhibition of PAH and ES uptake in hOAT1- or hOAT3-expressing oocytes, respectively. The IC(50) of steviol for hOAT1-mediated PAH transport was 11.1 microM compared with 62.6 microM for hOAT3-mediated ES uptake. The Michaelis-Menten inhibition constants (K(i)) for steviol transport mediated by hOAT1 and hOAT3 were 2.0 +/- 0.3 and 5.4 +/- 2.0 microM, respectively. Trans-stimulation of PAH efflux by steviol was assessed to determine whether steviol itself was transported by hOAT1 or hOAT3. A low concentration of 1 microM steviol increased the efflux of [(3)H]PAH (trans-stimulated) via both hOAT1 and hOAT3. In addition, it was shown by electrophysiology that steviol entry induced inward current in fOat1-expressing oocytes. In conclusion, stevioside had no interaction with either hOAT1 or hOAT3, whereas hOAT1, hOAT3, and fOat1 were all shown to be capable of steviol transport and thus, can play a role in its renal transport and excretion.


Multiple reversals of optical binding force in plasmonic disk-ring nanostructures with dipole-multipole Fano resonances.
We study the optical far-field and near-field characteristics, and the optical force effects of plasmonic disk-ring nanostructures. There are multiple Fano features resulting from the scattering interferences of the hybridized modes from the disk's dipole and the ring's higher-order modes. In particular, it is found that the optical binding force between the disk and the ring shows multiple sign reversals spectrally, from the dipole-quadrupole regime up to the dipole-decapole regime. We show that the zero-force points can be categorized into two types: the positive-to-negative ones resulting from the Fano dip and the negative-to-positive ones associated with the transitions between dipole-multipole modes. The multiple sign reversals of the optical forces are tunable by the geometrical size and gap of the disk and ring. Such characters make it possible to organize unusual optical matters from individual plasmonic nanoparticles.


Theoretical analysis of the influence of aerosol size distribution and physical activity on particle deposition pattern in human lungs.
A theoretical comparison of modeled particle depositions in the human respiratory tract was performed by taking into account different particle number and mass size distributions and physical activity in an urban environment. Urban-air data on particulate concentrations in the size range 10 nm-10 microm were used to estimate the hourly average particle number and mass size distribution functions. The functions were then combined with the deposition probability functions obtained from a computerized ICRP 66 deposition model of the International Commission on Radiological Protection to calculate the numbers and masses of particles deposited in five regions of the respiratory tract of a male adult. The man's physical activity and minute ventilation during the day were taken into account in the calculations. Two different mass and number size distributions of aerosol particles with equal (computed) <10 microm particle mass concentrations gave clearly different deposition patterns in the central and peripheral regions of the human respiratory tract. The deposited particle numbers and masses were much higher during the day (0700-1900) than during the night (1900-0700) because an increase in physical activity and ventilation were temporally associated with highly increased traffic-derived particles in urban outdoor air. In future analyses of the short-term associations between particulate air pollution and health, it would not only be important to take into account the outdoor-to-indoor penetration of different particle sizes and human time-activity patterns, but also actual lung deposition patterns and physical activity in significant microenvironments.


Hungry for your alanine: when liver depends on muscle proteolysis.
Fasting requires complex endocrine and metabolic interorgan crosstalk, which involves shifting from glucose to fatty acid oxidation, derived from adipose tissue lipolysis, in order to preserve glucose for the brain. The glucose-alanine (Cahill) cycle is critical for regenerating glucose. In this issue of JCI, Petersen et al. report on their use of an innovative stable isotope tracer method to show that skeletal muscle-derived alanine becomes rate controlling for hepatic mitochondrial oxidation and, in turn, for glucose production during prolonged fasting. These results provide new insight into skeletal muscle-liver metabolic crosstalk during the fed-to-fasting transition in humans.


Adaptive design optimization: a mutual information-based approach to model discrimination in cognitive science.
Discriminating among competing statistical models is a pressing issue for many experimentalists in the field of cognitive science. Resolving this issue begins with designing maximally informative experiments. To this end, the problem to be solved in adaptive design optimization is identifying experimental designs under which one can infer the underlying model in the fewest possible steps. When the models under consideration are nonlinear, as is often the case in cognitive science, this problem can be impossible to solve analytically without simplifying assumptions. However, as we show in this letter, a full solution can be found numerically with the help of a Bayesian computational trick derived from the statistics literature, which recasts the problem as a probability density simulation in which the optimal design is the mode of the density. We use a utility function based on mutual information and give three intuitive interpretations of the utility function in terms of Bayesian posterior estimates. As a proof of concept, we offer a simple example application to an experiment on memory retention.


DNA origami templated self-assembly of discrete length single wall carbon nanotubes.
Constructing intricate geometric arrangements of components is one of the central challenges of nanotechnology. Here we report a convenient, versatile method to organize discrete length single-walled carbon nanotubes (SWNT) into complex geometries using 2D DNA origami structures. First, a size exclusion HPLC purification protocol was used to isolate uniform length, SWNTs labelled with single stranded DNA (ssDNA). The nanotube-bound ssDNAs are composed of two domains: a SWNT binding domain and a linker binding domain. Although initially bound to the SWNTs, the linker domain is displaced from the surface by the addition of an external ssDNA linker strand. One portion of the linker strand is designed to form a double helix with the linker binding domain, compelling the DNA to project away from the SWNT surface. The remainder of the linker strand contains an ssDNA origami recognition sequence available for hybridization to a DNA origami nanostructure. Two different 2D DNA origami structures, a triangle and a rectangle, were used to organize the nanotubes. Several arrangements of nanotubes were constructed, with defined tube lengths and inter-tube angles. The uniform tube lengths and positional precision that this method affords may have applications in electronic device fabrication.


Isotope labeling and theoretical study of the formation of a3* ions from protonated tetraglycine.
Extensive 13C, 15N, and 2H labeling of tetraglycine was used to investigate the b3+ --> a3* reaction during low-energy collision-induced dissociation (CID) in a quadrupole ion-trap mass spectrometer. The patterns observed with respect to the retention or elimination of the isotope labels demonstrate that the reaction pathway involves elimination of CO and NH3. The ammonia molecule includes 2 H atoms from amide or amino positions, and one from an alpha-carbon position. The loss of NH3 does not involve elimination of the N-terminal amino group but, instead, the N atom of the presumed oxazolone ring in the b3+ ion. The CO molecule eliminated is the carbonyl group of the same oxazolone ring, and the alpha-carbon H atom is transferred from the amino acid adjacent to the oxazolone ring. Quantum chemical calculations indicate a multistep reaction cascade involving CO loss on the b3 --> a3 pathway and loss of NH=CH2 from the a3 ion to form b2. In the postreaction complex of b2 and NH=CH2, the latter can be attacked by the N-terminal amino group of the former. The product of this attack, an isomerized a3 ion, can eliminate NH3 from its N-terminus to form a3*. Calculations suggest that the ammonia and a3* species can form various ion-molecule complexes, and NH3 can initiate relay-type mobilization of the oxazolone H atoms from alpha-carbon positions to form a new oxazolone isomer. This multiple-step reaction scheme clearly explains the isotope labeling results, including unexpected scrambling of H atoms from alpha-carbon positions.


Galiellalactone induces cell cycle arrest and apoptosis through the ATM/ATR pathway in prostate cancer cells.
Galiellalactone (GL) is a fungal metabolite that presents antitumor activities on prostate cancer in vitro and in vivo. In this study we show that GL induced cell cycle arrest in G2/M phase, caspase-dependent apoptosis and also affected the microtubule organization and migration ability in DU145 cells. GL did not induce double strand DNA break but activated the ATR and ATM-mediated DNA damage response (DDR) inducing CHK1, H2AX phosphorylation (fH2AX) and CDC25C downregulation. Inhibition of the ATM/ATR activation with caffeine reverted GL-induced G2/M cell cycle arrest, apoptosis and DNA damage measured by fH2AX. In contrast, UCN-01, a CHK1 inhibitor, prevented GL-induced cell cycle arrest but enhanced apoptosis in DU145 cells. Furthermore, we found that GL did not increase the levels of intracellular ROS, but the antioxidant N-acetylcysteine (NAC) completely prevented the effects of GL on fH2AX, G2/M cell cycle arrest and apoptosis. In contrast to NAC, other antioxidants such as ambroxol and EGCG did not interfere with the activity of GL on cell cycle. GL significantly suppressed DU145 xenograft growth in vivo and induced the expression of fH2AX in the tumors. These findings identify for the first time that GL activates DDR in prostate cancer.


Sexual possibility situations and sexual behaviors among young adolescents: the moderating role of protective factors.
To examine sexual possibility situations (SPS) and protective practices associated with involvement in intimate sexual behaviors and the initiation of sexual intercourse among young adolescents and to determine if protective factors moderate the relationship between SPS and sexual behaviors. Data for these analyses were obtained from the baseline assessment for adolescents conducted as part of an HIV prevention study called "Keepin' it R.E.A.L.!" The study was conducted with a community-based organization (CBO) in an urban area serving a predominantly African-American population. In addition to items assessing SPS, intimate sexual behaviors, and initiation of sexual intercourse, adolescents provided information on the following protective factors: educational goals, self-concept, future time perspective, orientation to health, self-efficacy, outcome expectations, parenting, communication, values, and prosocial activities. Background personal information, including age and gender, was also collected. The analyses were conducted on data from 491 predominantly African-American adolescents, 61% of whom were boys. Variables were combined to form SPS and protective indices that were used in the first set of regression analyses. In a second set of analyses, the indices were unbundled and individual variables were entered into regression analyses. Both SPS and protective indices explained significant portions of variance in intimate sexual behaviors, and the SPS index explained a significant portion of variance in the initiation of sexual intercourse. The regression analysis using the unbundled SPS and protective factors revealed the following statistically significant predictors for intimate sexual behaviors: age, gender, time alone with groups of peers, time alone with a member of the opposite sex, behavior self-concept, popularity self-concept, self-efficacy for abstinence, outcome expectations for abstinence, parental control, personal values, and parental values. A similar regression analysis revealed that age, time alone with a member of the opposite sex, and personal values were significant predictors of initiation of sexual intercourse. These results provide evidence for the important role of protective factors in explaining early involvement in sexual behaviors and show that protective factors extend beyond personal characteristics to include both familial and peer factors.


Adult blood lead epidemiology and surveillance--United States, 1998-2001.
Elevated blood lead levels (BLLs) in adults can damage the cardiovascular, central nervous, reproductive, hematologic, and renal systems. The majority of cases are workplace-related. U.S. Department of Health and Human Services recommends that BLLs among all adults be reduced to < 25 microg/dL. The highest BLL acceptable by standards of the U.S. Occupational Safety and Health Administration is 40 microg/dL. The mean BLL of adults in the United States is < 3 microg/dL. This report covers cases of adults (aged > or = 16 years) with BLLs > or = 25 microg/dL, as reported by 25 states during 1998-2001. Since 1987, CDC has sponsored the state-based Adult Blood Lead Epidemiology and Surveillance (ABLES) program to track cases of elevated BLLs and provide intervention consultation and other assistance. Overall ABLES program data were last published in 1999 for the years 1994-1997. This report provides an update with data from 25 states reporting for > or = 2 years during 1998-2001. During that period, the ABLES program funded surveillance in 21 states - Alabama, Arizona, Connecticut, Iowa, Maryland, Massachusetts, Michigan, Minnesota, New Jersey, New York, North Carolina, Ohio, Oklahoma, Oregon, Pennsylvania, Rhode Island, South Carolina, Texas, Washington, Wisconsin, and Wyoming. Four additional states - California, Nebraska, New Hampshire, and Utah contributed data without CDC funding. During 1998-2001, the overall program's annual mean state prevalence rate for adults with BLLs > or = 25 microg/dL was 13.4/100,000 employed adults. This compares with 15.2/100,000 for 1994-1997. Yearly rates were 13.8 (1998), 12.9 (1999), 14.3 (2000), and 12.5 (2001). For adults with BLLs > or = 40 microg/dL, the overall program's annual mean state prevalence rare during 1998-2001 was 2.9/ 100,000 employed adults. This compares with 3.9/100,000 for 1994-1997. Yearly rates were 3.3 (1998), 2.5 (1999), 2.9 (2000), and 2.8 (2001). Although certain limitations exist, the overall ABLES data indicate a declining trend in elevated BLLs among employed adults. ABLES-funded states increased from 21 to 35 in 2002, and more detailed reporting requirements were put into effect. These, and other improvements, will enable the ABLES program to work more effectively toward its 2010 target of eliminating all cases of BLLs > or = 25 microg/dL in adults caused by workplace exposures.


Neurocognitive function of emergency department patients with mild traumatic brain injury.
We characterize the neurocognitive function of patients presenting to the emergency department (ED) with mild traumatic brain injury. This prospective study took place at an urban, academic ED and Level I trauma center. Case patients consisted of a convenience sample of ED patients aged 18 to 59 years, presenting to the ED with mild traumatic brain injury and having a head computed tomography scan without traumatic abnormalities. Controls consisted of patients aged 18 to 59 years, presenting to the ED with an isolated, nondominant hand extremity injury. We excluded patients with multiple injuries and recent alcohol consumption. Subjects completed a computerized neurocognitive test battery (Immediate Post-concussion Assessment and Cognitive Testing). The primary measures were verbal memory, visual memory, and visual motor and reaction speed. We compared raw and age-normalized neurocognitive performance between case patients and controls by using nonparametric statistics. We included a total of 23 head-injured case patients and 31 non-head-injured controls. Case patients and controls exhibited similar raw (median 80.1 versus 85.0 points; difference in medians -4.9; P=.26) and age-normalized (31.9 versus 57.4 percentile; difference in medians -25.5; P=.12) verbal memory. Case patients and controls exhibited similar raw (64.6 versus 63.5; difference 1.1; P=.79) and age-normalized (20.8 versus 25.8 percentile; difference -5.0; P=.44) visual memory. Compared with controls, mild traumatic brain injury case patients demonstrated slower raw (31.6 versus 37.0 points; difference -5.4; P=.002) and age-normalized (17.1 versus 57.6 percentile; difference -40.5; P=.001) visual motor speed. Mild traumatic brain injury case patients exhibited slower raw (median 0.66 versus 0.60 seconds; difference 0.06; P=.01) and age-normalized (29.3 versus 42.8 percentile; difference -13.5; P=.009) reaction times. In conclusion, compared with the non-head-injured patients, ED mild traumatic brain injury patients demonstrated subtle but discernible neurocognitive deficits.


First Report of Pepper mottle virus Infecting Tomato in Hawaii.
In August 2011, tomato (Solanum lycopersicum L.) fruit from a University of Hawaii field trial displayed mottling symptoms similar to that caused by Tomato spotted wilt virus (TSWV) or other tospoviruses. The foliage from affected plants, however, appeared symptomless. Fruit and leaf tissue from affected plants were negative for TSWV analyzed by double antibody sandwich (DAS)-ELISA and/or TSWV ImmunoStrips (Agdia, Elkhart, IN) when performed following the manufacturer's instructions. Total RNA from a symptomatic and an asymptomatic plant was isolated using an RNeasy Plant Mini Kit (Qiagen, Valencia, CA) and reverse transcribed using Invitrogen SuperScript III reverse transcriptase (Life Technologies, Grand Island, NY) and primer 900 (5'- CACTCCCTATTATCCAGG(T)16-3') following the enzyme manufacturer's instructions. The cDNA was then used as template in a universal potyvirus PCR assay using primers 900 and Sprimer, which amplify sequences encoding the partial inclusion body protein (NIb), coat protein, and 3' untranslated region of potyviruses (1). A ~1,700-bp product was amplified from the cDNA of the symptomatic plant but not the asymptomatic plant. This product was cloned using pGEM-T Easy (Promega, Madison, WI) and three clones were sequenced at the University of Hawaii's Advanced Studies in Genomics, Proteomics, and Bioinformatics laboratory. The 1,747-bp consensus sequence of the three clones was deposited in GenBank (Accession No. JQ429788) and, following primer sequence trimming, found to be 97% identical to positions 7,934 through 9,640 of Pepper mottle virus (PepMoV; family Potyviridae, genus Potyvirus) accessions from Korea (isolate '217' from tomato; EU586126) and California (isolate 'C' from pepper; M96425). To determine the incidence of PepMoV in the field trial, all 292 plants representing 14 tomato cultivars were assayed for the virus 17 weeks after planting using a PepMoV-specific DAS-ELISA (Agdia) following the manufacturer's directions. Plants were considered positive if their mean absorbance at 405 nm was greater than the mean absorbance + 3 standard deviations + 10% of the negative control samples. The virus incidence ranged from 4.8 to 47.6% for the different varieties, with an overall incidence of 19.9%. Although plant growth was not noticeably impaired by PepMoV infection, the majority of fruit from infected plants was unsaleable, making PepMoV a considerable threat to tomato production in Hawaii. PepMoV has been reported to naturally infect tomato in Guatemala (3) and South Korea (2). To our knowledge, this is the first report of this virus in Hawaii and the first report of this virus naturally infecting tomato in the United States. References: (1) J. Chen et al. Arch. Virol. 146:757, 2001. (2) M.-K. Kim et al. Plant Pathol. J. 24:152, 2008. (3) J. Th. J. Verhoeven et al. Plant Dis. 86:186, 2002.


Occurrence of Acetamiprid Residues in Water Reservoirs in the Cotton Basin of Northern Benin.
An Ultra performance liquid chromatography (UPLC) coupled to UV detection method was developed to determine acetamiprid residues in water reservoirs of northern Benin, close to cotton fields. The quantification limit of this method was 0.2 µg L-1 acetamiprid in water, its precision ranged between 8% and 22%, and its trueness between 99% and 117% (for concentrations ranging from 0.2 to 5.0 µg L-1). Acetamiprid residues were determined in water samples collected in four reservoirs from northern Benin during the phytosanitary treatment period of cotton. The minimum and maximum concentrations of acetamiprid residues in water were 0.2 and 7.7 µg L-1, respectively. These levels do not represent any risk for human consumption of this water, but indicate a regular use of acetamiprid, possibly together with other pesticides which could be more harmful for both humans and aquatic species.


Pain and activity after vaginal reconstructive surgery for pelvic organ prolapse and stress urinary incontinence.
Little is known about short- and long-term pain and functional activity after surgery for pelvic organ prolapse. The objectives of the study were to describe postoperative pain and functional activity after transvaginal native tissue reconstructive surgery with apical suspension and retropubic synthetic midurethral sling and to compare these outcomes between patients receiving 2 common transvaginal prolapse repairs, uterosacral ligament, and sacrospinous ligament vaginal vault suspension. This planned secondary analysis of a 2 × 2 factorial randomized trial included 374 women randomized to receive uterosacral (n = 188) or sacrospinous (n = 186) vaginal vault suspension to treat both stages 2-4 apical vaginal prolapse and stress urinary incontinence between 2008 and 2013 at 9 medical centers. Participants were also randomized to receive perioperative pelvic muscle therapy or usual care. All patients received transvaginal native tissue repairs and a midurethral sling. Participants completed the Surgical Pain Scales (0-10 numeric rating scales; higher scores = greater pain) and Activity Assessment Scale (0-100; higher score = higher activity) prior to surgery and at 2 weeks, 4-6 weeks, and 3 months postoperatively. The MOS 36-item Short-Form Health Survey was completed at baseline and 6, 12, and 24 months after surgery; the bodily pain, physical functioning, and role-physical subscales were used for this analysis (higher scores = less disability). Self-reported pain medication use was also collected. Before surgery, average pain at rest and during normal activity were (adjusted mean ± SE) 2.24 ± 0.23 and 2.76 ± 0.25; both increased slightly from baseline at 2 weeks (+0.65, P = .004, and +0.74, P = .007, respectively) and then decreased below baseline at 3 months (-0.87 and -1.14, respectively, P < .001), with no differences between surgical groups. Pain during exercise/strenuous activity and worst pain decreased below baseline levels at 4-6 weeks (-1.26, P = .014, and -0.95, P = .002) and 3 months (-1.97 and -1.50, P < .001) without differences between surgical groups. Functional activity as measured by the Activity Assessment Scale improved from baseline at 4-6 weeks (+9.24, P < .001) and 3 months (+13.79, P < .001). The MOS 36-item Short-Form Health Survey Bodily Pain, Physical Functioning, and Role-Physical Scales demonstrated significant improvements from baseline at 6, 12, and 24 months (24 months: +5.62, +5.79, and +4.72, respectively, P < .001 for each) with no differences between groups. Use of narcotic pain medications was reported by 14.3% of participants prior to surgery and 53.7% at 2 and 26.1% at 4-6 weeks postoperatively; thereafter use was similar to baseline rates until 24 months when it decreased to 6.8%. Use of nonnarcotic pain medication was reported by 48.1% of participants prior to surgery, 68.7% at 2 weeks, and similar to baseline at 3 months; thereafter use dropped steadily to 26.6% at 2 years. Uterosacral ligament suspension resulted in less new or worsening buttock pain than sacrospinous suspension at 4-6 weeks postoperatively (4.6% vs 10.5%, P = .043) but no difference in groin or thigh pain. Pain and functional activity improve for up to 2 years after native tissue reconstructive surgery with uterosacral or sacrospinous vaginal vault suspension and midurethral sling for stages 2-4 pelvic organ prolapse. On average, immediate postoperative pain is low and improves to below baseline levels by 4-6 weeks.


Familial aggregation of mucosal leishmaniasis in northeast Brazil.
To evaluate whether familial clustering occurs in mucosal leishmaniasis (ML), patients with ML (index cases) were randomly selected from medical records at a health post in an endemic area for Leishmania braziliensis infection. Control individuals (index controls) matched by age, gender, and place of residence to index cases were selected. Family members of index cases and controls were compared with respect to environmental factors and the incidence of cutaneous leishmaniasis (CL) and ML. Delayed type hypersensitivity test (DTH) to Leishmania antigen was tested in selected families. Among 289 members of 46 families enrolled, significant differences were found in the frequencies of CL (37% versus 20%) and ML (5% versus 0) in case versus control families, respectively. Families with 2 cases of ML had a higher frequency (29.6%) of DTH-positive individuals than control families (9.4%). These data demonstrate familial clustering of CL, ML, and positive DTH skin tests in an area endemic for L. braziliensis infection.


Pulmonary hilar lymph node metastasis of breast cancer induced bronchopleural fistula and superior vena cava syndrome.
It is extremely rare for pulmonary hilar lymph node metastasis (PHLNM) of a cancer to be independently lethal. Here, we report an exceedingly rare case of cavitation in PHLNM from breast cancer triggering broncho-pleural fistula and empyema (BPFE), complicated with superior vena cava syndrome (SVCS). A 56-year-old woman who had undergone left segmental mastectomy and axillary lymph node dissection due to left breast cancer was then treated for 1 year with postoperative adjuvant chemotherapy. Recurrence of right PHLNM was observed 2 years after the operation, for which 3 courses of bevacizumab (BEV) and paclitaxel combination chemotherapy were administered. The woman had dyspnea and fever during the washout period, and CT examination revealed fistula formation between the right PHLNM cavitation and right main bronchus, so she was admitted for further treatment. This fistula rapidly progressed to BPFE, and contralateral aspiration was observed to cause pneumonia of the left lung. In addition, edema of both upper limbs and head and neck were observed, and CT examination revealed SVCS caused by re-enlargement of PHLNM. Active treatment was performed, but the recommencement of chemotherapy was not possible, and she died on Day 150 of admission. We think that PHLNM deteriorated to central necrosis due to chemotherapy with BEV taking effect, leading to formation of BPFE. The case was also made more difficult due to the complication of SVCS caused by the re-enlarged PHLNM.


Diastolic function of the heart in clinical cardiology.
During the last six years, there has been increased interest in the detection of abnormalities of left ventricular diastolic function in patients with heart disease. Before 1981, most studies on diastolic function were performed in the catheter laboratory using invasive techniques and complex methods. Recently, radionuclide angiograms and Doppler echocardiography have been employed to measure the dynamics of filling in normal individuals and in patients with heart disease. These methods are noninvasive, easy to perform, accurate, and reproducible. It is now clear that diastolic function may be altered globally and regionally, at rest and perhaps during exercise, in many patients with ischemic heart disease, hypertension, and hypertrophic cardiomyopathy. Interestingly, these diastolic abnormalities may even appear before systolic abnormalities are identified in these patients. Thus, diastolic abnormalities may permit assessment of presence of disease early in its evolution. Whether detection and quantitation of diastolic abnormalities will permit grading of disease severity or evaluation of therapeutic efficacy remains an important research question. At the present time, it appears that the decision to employ either radionuclide angiography or Doppler echocardiography for the assessment of diastolic abnormalities will depend on the local expertise to carry out the investigation. Both diagnostic modalities require standardization of accuracy and reproducibility with proper selection of control values from the appropriate populations of normal individuals. It is also important to remember that left ventricular diastolic abnormalities have to be identified after the elimination of the confounding influence of variables such as ejection fraction, heart rate, age, and preload (end-diastolic volume). Automation of the derivation of indexes of diastolic filling should provide an objective assessment of the dynamics of left ventricular filling. Although the value of measurement of diastolic filling in the individual patient remains controversial, we believe that the practice of cardiology is incomplete without consideration of the second half of cardiac function.


A binary image reconstruction technique for accurate determination of the shape and location of metal objects in x-ray computed tomography.
The presence of metals in patients causes streaking artifacts in X-ray CT and has been recognized as a problem that limits various applications of CT imaging. Accurate localization of metals in CT images is a critical step for metal artifacts reduction in CT imaging and many practical applications of CT images. The purpose of this work is to develop a method of auto-determination of the shape and location of metallic object(s) in the image space. The proposed method is based on the fact that when a metal object is present in a patient, a CT image can be divided into two prominent components: high density metal and low density normal tissues. This prior knowledge is incorporated into an objective function as the regularization term whose role is to encourage the solution to take a form of two intensity levels. A computer simulation study and four experimental studies are performed to evaluate the proposed approach. Both simulation and experimental studies show that the presented algorithm works well even in the presence of complicated shaped metal objects. For a hexagonally shaped metal embedded in a water phantom, for example, it is found that the accuracy of metal reconstruction is within sub-millimeter.


Birds from the Azores: An updated list with some comments on species distribution.
An updated checklist of the Birds of the Azores is presented based on information compiled from Rodrigues et al. (2010) and from the websites, Azores Bird Club. (2014), Aves dos Açores (2014) Azores Bird Sightings (2014) and Vittery (2014), since 2010. The checklist has a total of 414 species, including 38 new species. Almost half of the species and subspecies that occur in the Azores have a Palearctic origin, the remaining ones being essentialy Nearctic and Holarctic species. São Miguel is the island with the highest number of bird species, followed by Terceira, Corvo and Flores islands.


COVID-19-Related Pleural Diseases.
Coronavirus disease 2019 (COVID-19)-related pleural diseases are now well recognized. Since the beginning of the pandemic, increasing cases of pleural diseases including pneumothorax, pneumomediastinum, and pleural effusion with severe COVID-19 infection have attracted the attention of physicians and are not incidental or due to barotrauma. The complicated course of COVID-19 illness highlights the complex pathophysiological underpinnings of pleural complications. The management of patients with pneumothorax and pneumomediastinum is challenging as the majority require assisted ventilation; physicians therefore appear to have a low threshold to intervene. Conversely, pleural effusion cases, although sharing some similar patient characteristics with pneumothorax and pneumomediastinum, are in general managed more conservatively. The evidence suggests that patients with COVID-19-related pleural diseases, either due to air leak or effusion, have more severe disease with a worse prognosis. This implies that prompt recognition of these complications and targeted management are key to improve outcomes.


Change in joint space width: hyaline articular cartilage loss or alteration in meniscus?
To explore the relative contribution of hyaline cartilage morphologic features and the meniscus to the radiographic joint space. The Boston Osteoarthritis of the Knee Study is a natural history study of symptomatic knee osteoarthritis (OA). Baseline and 30-month followup assessments included knee magnetic resonance imaging (MRI) and fluoroscopically positioned weight-bearing knee radiographs. Cartilage and meniscal degeneration were scored on MRI in the medial and lateral tibiofemoral joints using a semiquantitative grading system. Meniscal position was measured to the nearest millimeter. The dependent variable was joint space narrowing (JSN) on the plain radiograph (possible range 0-3). The predictor variables were MRI cartilage score, meniscal degeneration, and meniscal position measures. We first conducted a cross-sectional analysis using multivariate regression to determine the relative contribution of meniscal factors and cartilage morphologic features to JSN, adjusting for body mass index (BMI), age, and sex. The same approach was used for change in JSN and change in predictor variables. We evaluated 264 study participants with knee OA (mean age 66.7 years, 59% men, mean BMI 31.4 kg/m(2)). The results from the models demonstrated that meniscal position and meniscal degeneration each contributed to prediction of JSN, in addition to the contribution by cartilage morphologic features. For change in medial joint space, both change in meniscal position and change in articular cartilage score contributed substantially to narrowing of the joint space. The meniscus (both its position and degeneration) accounts for a substantial proportion of the variance explained in JSN, and the change in meniscal position accounts for a substantial proportion of change in JSN.


Unraveling the Composition of the Root-Associated Bacterial Microbiota of Phragmites australis and Typha latifolia.
Phragmites australis and Typha latifolia are two macrophytes commonly present in natural and artificial wetlands. Roots of these plants engage in interactions with a broad range of microorganisms, collectively referred to as the microbiota. The microbiota contributes to the natural process of phytodepuration, whereby pollutants are removed from contaminated water bodies through plants. The outermost layer of the root corpus, the rhizoplane, is a hot-spot for these interactions where microorganisms establish specialized aggregates designated biofilm. Earlier studies suggest that biofilm-forming members of the microbiota play a crucial role in the process of phytodepuration. However, the composition and recruitment cue of the Phragmites, and Typha microbiota remain poorly understood. We therefore decided to investigate the composition and functional capacities of the bacterial microbiota thriving at the P. australis and T. latifolia root-soil interface. By using 16S rRNA gene Illumina MiSeq sequencing approach we demonstrated that, despite a different composition of the initial basin inoculum, the microbiota associated with the rhizosphere and rhizoplane of P. australis and T. latifolia tends to converge toward a common taxonomic composition dominated by members of the phyla Actinobacteria, Firmicutes, Proteobacteria, and Planctomycetes. This indicates the existence of a selecting process acting at the root-soil interface of these aquatic plants reminiscent of the one observed for land plants. The magnitude of this selection process is maximum at the level of the rhizoplane, where we identified different bacteria enriched in and discriminating between rhizoplane and rhizosphere fractions in a species-dependent and -independent way. This led us to hypothesize that the structural diversification of the rhizoplane community underpins specific metabolic capabilities of the microbiota. We tested this hypothesis by complementing the sequencing survey with a biochemical approach and scanning electron microscopy demonstrating that rhizoplane-enriched bacteria have a bias for biofilm-forming members. Together, our data will be critical to facilitate the rational exploitation of plant-microbiota interactions for phytodepuration.


Insulin Resistance as Common Molecular Denominator Linking Obesity to Alzheimer's Disease.
Alzheimer's disease (AD) is an aging-related multi-factorial disorder to which metabolic factors contribute at what has canonically been considered a centrally mediated process. Although the exact underlying mechanisms are still unknown, obesity is recognized as a risk factor for AD and the condition of insulin resistance seems to be the link between the two pathologies. Using mice with high fat diet (HFD) obesity we dissected the molecular mechanisms shared by the two disorders. Brains of HFD fed mice showed elevated levels of APP and Aβ40/Aβ42 together with BACE, GSK3β and Tau proteins involved in APP processing and Aβ accumulation. Immunofluorescence, Thioflavin T staining experiments, confirmed increased Aβ generation, deposition in insoluble fraction and plaques formation in both the hippocampus and the cerebral cortex of HFD mice. Presence of Aβ40/Aβ42 in the insoluble fraction was also shown by ELISA assay. Brain insulin resistance was demonstrated by reduced presence of insulin receptor (IRs) and defects in Akt-Foxo3a insulin signaling. We found reduced levels of phospho-Akt and increased levels of Foxo3a in the nuclei of neurons where proapototic genes were activated. Dysregulation of different genes related to insulin resistance, especially those involved in inflammation and adipocytokines synthesis were analyzed by Profiler PCR array. Further, HFD induced oxidative stress, mitochondrial dysfunction and dynamics as demonstrated by expression of biomarkers involved in these processes. Here, we provide evidence that obesity and AD markers besides insulin resistance are associated with inflammation, adipokine dyshomeostasis, oxidative stress and mitochondrial dysfunction, all mechanisms leading to neurodegeneration.


Integrated Evaluation of Urban Development Suitability Based on Remote Sensing and GIS Techniques - A Case Study in Jingjinji Area, China.
Jingjinji area (namely Beijing, Tianjin and He Bei Province) is one of the three largest regional economic communities in China. Urban expansion has sped up in the past 20 years in this area due to the rapid economic and population growth. Evaluating the landuse suitability for urban growth on a regional scale is an urgent need, because the most suitable areas and the most suitable scale of urban growth can thus be determined accordingly. In order to meet this requirement, remote sensing and geographic information system (GIS) techniques were adopted, and an integrated evaluating model was developed supported by AHP method. The integrated urban development suitability index (UDSI) was calculated using this model. According to the UDSI result, the spatial distribution of urban development suitability and its driving forces were analyzed. Urban boundaries in 1995, 2000 and 2005, which were derived from Landsat TM/ETM+ satellite data, were overlaid on the UDSI map, and the suitable urban develop tendency in this area were discussed. The result of this study indicated that integrated evaluation of urban development could be conducted in an operational way using remote sensing data, GIS spatial analysis technique and AHP modeling method.


[Relationship between physiological parameters changes and severe heatstroke induced by 5-km armed cross-country training].
To explore the relationship between physiological parameters changes and severe heatstroke induced by 5-km armed cross-country training. A total of 521 male officers and soldiers from a special team who participated in the summer training of 5-km armed cross-country training from year 2016 to 2017 were enrolled. All trainees participated in 5-km armed cross-country training in high temperature and humidity environment of ambient temperature > 32 centigradeand (or) relative humidity > 65%. The trainees were divided into two groups according to the incidence of severe heatstroke in the course of training. The age, enlistment time, constitution score, body mass index (BMI), external environment (ambient temperature, relative humidity, wind speed, heat index) of trainees of the two groups, and the change rates of arterial blood oxygen saturation (SaO2), body temperature, pulse and blood pressure within 5 minutes after the 5-km armed cross-country training were compared between the two groups. The risk factors of severe heatstroke were screened by two classified Logistic regression analysis, and the predictive value of various risk factors of severe heatstroke was analyzed by the receiver operator characteristic curve (ROC). In 521 trainees of 5-km armed cross-country training, 29 trainees suffered from severe heatstroke accounting for 5.57%. There was no significant difference in the age, enlistment time, constitution score, BMI, or external environment during 5-km armed cross-country training between severe heatstroke group and non-severe heatstroke group. Compared with those without severe heatstroke, the descending rates of body temperature, pulse, blood pressure and SaO2 increased rate within 5 minutes after 5-km armed cross-country training of severe heatstroke trainees were significantly decreased [temperature descending rate: (0.67±0.30)% vs. (1.43±1.28)%, pulse descending rate: (7.53±5.21)% vs. (13.48±8.07)%, blood pressure descending rate: (9.28±6.84)% vs. (19.42±7.73)%, SaO2 increased rate: (0.51±0.39)% vs. (1.50±1.43)%, all P < 0.01]. Two classification Logistic regression analysis showed that the temperature descending rate [odds ratio (OR) = 0.485, 95% confidence interval (95%CI) = 0.289-0.817], pulse descending rate (OR = 0.903, 95%CI = 0.845-0.965), blood pressure descending rate (OR = 0.841, 95%CI = 0.790-0.896), and SaO2 increased rate (OR = 0.421, 95%CI = 0.250-0.711) were the risk factors for severe heatstroke during 5-km armed cross-country training (all P < 0.01). ROC curve analysis showed that temperature descending rate [area under ROC curve (AUC) = 0.659, 95%CI = 0.604-0.714], pulse descending rate (AUC = 0.730, 95%CI = 0.762-0.900), blood pressure descending rate (AUC = 0.831, 95%CI = 0.659-0.801), SaO2 increased rate (AUC = 0.711, 95%CI = 0.655-0.767) could be used for the incidence of severe heatstroke prediction during 5-km armed cross-country training (all P < 0.01), and the predicted value was the same. Under the same conditions, the severe heatstroke during 5-km cross-country training is closely related to the descending rates of body temperature, pulse, and blood pressure as well as SaO2 increased rate within 5 minutes after the training, whose predictive values for severe heatstroke were the same.


Antimicrobial Resistance in the Asia Pacific region: a meeting report.
The Asia Pacific region, home to two-thirds of the world's population and ten of the least developed countries, is considered a regional hot-spot for the emergence and spread of antimicrobial resistance (AMR). Despite this, there is a dearth of high-quality regional data on the extent of AMR. Recognising the urgency to close this gap, Singapore organised a meeting to discuss the problems in the region and frame a call for action. Representatives from across the region and beyond attended the meeting on the "Antimicrobial Resistance in the Asia Pacific & its impact on Singapore" held in November 2018. This meeting report is a summary of the discussions on the challenges and progress in surveillance, drivers and levers of AMR emergence, and the promising innovations and technologies that could be used to combat the increasing threat of AMR in the region. Enhanced surveillance and research to provide improved evidence-based strategies and policies are needed. The major themes that emerged for an action plan are working towards a tailored solution for the region by harnessing the One Health approach, enhancing inter-country collaborations, and collaboratively leverage upon new emerging technologies. A regionally coordinated effort that is target-driven, sustainable and builds on a framework facilitating communication and governance will strengthen the fight against AMR in the Asia Pacific region.


k-t2 BLAST: exploiting spatiotemporal structure in simultaneously cardiac and respiratory time-resolved volumetric imaging.
Multidimensional imaging resolving both the cardiac and respiratory cycles simultaneously has the potential to describe important physiological interdependences between the heart and pulmonary processes. A fully five-dimensional acquisition with three spatial and two temporal dimensions is hampered, however, by the long acquisition time and low spatial resolution. A technique is proposed to reduce the scan time substantially by extending the k-t BLAST framework to two temporal dimensions. By sampling the k-t space sparsely in a lattice grid, the signal in the transform domain, x-f space, can be densely packed, exploiting the fact that large regions in the field of view have low temporal bandwidth. A volumetric online prospective triggering approach with full cardiac and respiratory cycle coverage was implemented. Retrospective temporal interpolation was used to refine the timing estimates for the center of k-space, which is sampled for all cardiac and respiratory time frames. This resulted in reduced reconstruction error compared with conventional k-t BLAST reconstruction. The k-t(2) BLAST technique was evaluated by decimating a fully sampled five-dimensional data set, and feasibility was further demonstrated by performing sparsely sampled acquisitions. Compared to the fully sampled data, a fourfold improvement in spatial resolution was accomplished in approximately half the scan time.


Non-neutral evolution and reciprocal monophyly of two expressed Mhc class II B genes in Leach's storm-petrel.
The major histocompatibility complex (Mhc) is subject to pathogen-mediated balancing selection and can link natural selection with mate choice. We characterized two Mhc class II B loci in Leach's storm-petrel, Oceanodroma leucorhoa, focusing on exon 2 which encodes the portion of the protein that binds pathogen peptides. We amplified and sequenced exon 2 with locus-specific nested PCR and Illumina MiSeq using individually barcoded primers. Repeat genotyping of 78 single-locus genotypes produced identical results in 77 cases (98.7%). Sequencing of messenger RNA (mRNA) from three birds confirmed expression of both loci, consistent with the observed absence of stop codons or frameshifts in all alleles. In 48 birds, we found 9 and 12 alleles at the two loci, respectively, and all 21 alleles translated to unique amino acid sequences. Unlike many studies of duplicated Mhc genes, alleles of the two loci clustered into monophyletic groups. Consistent with this phylogenetic result, interlocus gene conversion appears to have affected only two short fragments of the exon. As predicted under a paradigm of pathogen-mediated selection, comparison of synonymous and non-synonymous substitution rates found evidence of a history of positive selection at putative peptide binding sites. Overall, the results suggest that the gene duplication event leading to these two loci is not recent and that point mutations and positive selection on the peptide binding sites may be the predominant forces acting on these genes. Characterization of these loci sets the stage for population-level work on the evolutionary ecology of Mhc in this species.


Dementia and depression among the elderly living in the Hobart community: the effect of the diagnostic criteria on the prevalence rates.
A survey was made of 274 non-institutionalized persons aged 70 and over living in Hobart. The prevalence of dementia and of depression was measured by interviewing subjects using a modified version of the Geriatric Mental State Schedule (GMS) (Copeland et al. 1976) and the Mini Mental State Examination (MMSE) (Folstein et al. 1975). Rates of morbidity were derived from different diagnostic procedures. These were: diagnoses made by a psychiatrist (A.S.H.) directly from the interview schedules and audiotapes, and rated as mild, moderate or severe; the criteria laid down in DSM-III, converted into algorithms describing 3 degrees of severity; and the algorithms for pervasive dementia and depression proposed by Gurland et al. (1983), and from these authors' rational scales. In addition, the relation between scales for dementia and for depression and the diagnosed categories was examined. Some problems in applying these methods to aged persons in the community are discussed. It is concluded that more detailed specification of criteria is desirable if the comparative epidemiology of dementia and depression in old age is to advance.


Fosfomycin Resistance in Escherichia coli Isolates from South Korea and in vitro Activity of Fosfomycin Alone and in Combination with Other Antibiotics.
We investigated fosfomycin susceptibility in Escherichia coli clinical isolates from South Korea, including community-onset, hospital-onset, and long-term care facility (LTCF)-onset isolates. The resistance mechanisms and genotypes of fosfomycin-resistant isolates were also identified. Finally, the in vitro efficacy of combinations of fosfomycin with other antibiotics were examined in susceptible or extended spectrum β-lactamase (ESBL)-producing E. coli isolates. The fosfomycin resistance rate was 6.7% and was significantly higher in LTCF-onset isolates than community-onset and hospital-onset isolates. Twenty-one sequence types (STs) were identified among 19 fosfomycin-resistant E. coli isolates, showing diverse genotypes. fosA3 was found in only two isolates, and diverse genetic variations were identified in three genes associated with fosfomycin resistance, namely, GlpT, UhpT, and MurA. Some fosfomycin-resistant E. coli isolates carried no mutations. In vitro time-kill assays showed that fosfomycin alone did not exhibit an excellent killing activity, compared with ciprofloxacin in susceptible isolates and with ertapenem in ESBL producers. However, combining fosfomycin with cefixime or piperacillin-tazobactam eradicated susceptible or ESBL-producing isolates, respectively, even with 0.5× minimum inhibitory concentrations. Overall, we found a relatively high fosfomycin resistance rate in E. coli isolates from South Korea. Based on their genotypes and resistance mechanisms, most of the fosfomycin-resistant E. coli isolates might occur independently. Antibiotic combinations with fosfomycin could be a suitable therapeutic option for infections caused by E. coli isolates.


HIV-1 seroconversion incidence following pregnancy and delivery among women seronegative at recruitment in Harare, Zimbabwe.
To estimate the incidence of HIV seroconversion among women following pregnancy and delivery. A prospective cohort of women who were HIV negative at recruitment on first antenatal care visit. Pregnant women were invited to undergo voluntary confidential HIV counselling and blood draw for HIV testing during the first antenatal care visit as part of a prospective study of mother-to-child transmission of HIV-1. Repeat tests were conducted at delivery, six weeks post partum and at three monthly intervals until 24 months or on termination due to subsequent pregnancy, death or loss to follow up. Logistic regression modelling was used to determine independent predictors of HIV seroconversion. Among 372 HIV negative pregnant women who were enrolled, 66 seroconverted during follow up, resulting in a sero-incidence of 4.8 per 100 person years (95% confidence interval [CI], 3.1 to 6.5). Women who did not seroconvert during the time of pregnancy or follow up were significantly more likely to have used a condom with their partners (OR = 0.68, 95% CI = 0.47 to 0.99). Women aged 17 years and below had the highest seroconversion incidence (6.25%) followed by those aged 18 to 19 years (5.42%). Women who seroconverted and those who were HIV positive at recruitment were more likely to be married. Lack of education by the partner of a pregnant woman constituted a significant risk factor for HIV seroconversion (OR = 2.8; 95% CI = 1.1 to 11.0). There is a high HIV seroconversion incidence among women during pregnancy and following delivery, especially those aged 19 years and below. Being married does not protect the women from the risk of HIV seroconversion. Strategies for HIV prevention should target pregnant women and their partners.


Early detection of subjects at risk for vascular remodelling - results from the Swiss population-based study SAPALDIA.
As the burden of cardiovascular disease (CVD) increases globally, its prevention and risk assessment becomes ever more important. We thus investigated the longitudinal association of the cardiovascular risk scores in the population-based cohort SAPALDIA with carotid intima media thickening (CIMT), an indicator of sub-clinical disease, and CVD incidence. In 2,832 SAPALDIA participants, the Swiss and ESC heart risk score (AGLA, SCORE) were calculated based on 2001 data and CIMT was measured in 2010/11. We ran multi-level linear regression analyses between scores and CIMT, stratified for CVD status and gender, and logistic analyses for doctor-diagnosed CVD incidence. Path analyses investigated direct and indirect effects on CIMT. AGLA and SCORE were positively associated with increasing CIMT in both healthy and CVD diagnosed subjects and men and women. Participants in highest risk categories showed a significant CIMT difference of >0.20 mm compared to the reference risk category (<1%), even larger in CVD healthy subjects and men. With increasing risk the odds of CVD incidence increased (Ref. <1%; 10 yr. risk AGLA >10% OR 2.1, >20% OR 3.7). Path analyses yield risk factors' direct and indirect effects through blood pressure. The positive longitudinal association between risk estimations and CIMT confirms the use of risk scores in assessing individuals and populations at risk. Systolic blood pressure appears to be a main pathological mechanism, underscoring the importance of optimal blood pressure control and the importance of prevention strategies of risk factors, indirectly affecting CIMT through the haemodynamic pathway.


Genomic analysis of wild tomato introgressions determining metabolism- and yield-associated traits.
With the aim of determining the genetic basis of metabolic regulation in tomato fruit, we constructed a detailed physical map of genomic regions spanning previously described metabolic quantitative trait loci of a Solanum pennellii introgression line population. Two genomic libraries from S. pennellii were screened with 104 colocated markers from five selected genomic regions, and a total of 614 bacterial artificial chromosome (BAC)/cosmids were identified as seed clones. Integration of sequence data with the genetic and physical maps of Solanum lycopersicum facilitated the anchoring of 374 of these BAC/cosmid clones. The analysis of this information resulted in a genome-wide map of a nondomesticated plant species and covers 10% of the physical distance of the selected regions corresponding to approximately 1% of the wild tomato genome. Comparative analyses revealed that S. pennellii and domesticated tomato genomes can be considered as largely colinear. A total of 1,238,705 bp from both BAC/cosmid ends and nine large insert clones were sequenced, annotated, and functionally categorized. The sequence data allowed the evaluation of the level of polymorphism between the wild and cultivated tomato species. An exhaustive microsynteny analysis allowed us to estimate the divergence date of S. pennellii and S. lycopersicum at 2.7 million years ago. The combined results serve as a reference for comparative studies both at the macrosyntenic and microsyntenic levels. They also provide a valuable tool for fine-mapping of quantitative trait loci in tomato. Furthermore, they will contribute to a deeper understanding of the regulatory factors underpinning metabolism and hence defining crop chemical composition.


Adherence to a Mediterranean-style eating pattern and risk of diabetes in a U.S. prospective cohort study.
A Mediterranean-style eating pattern is consistently associated with a decreased diabetes risk in Mediterranean and European populations. However, results in U.S. populations are inconsistent. The objective of this study was to assess whether a Mediterranean-style eating pattern would be associated with diabetes risk in a large, nationally representative U.S. cohort of black and white men and women. Participants from the Atherosclerosis Risk in Communities study prospective cohort without diabetes, cardiovascular disease, or cancer at baseline (visit 1, 1987-1989; n = 11,991) were included (mean age 54 years, 56% female, 75% white). Alternate Mediterranean Diet scores (aMed) were calculated using the mean dietary intake self-reported at visit 1 and visit 3 (1993-1995) or visit 1 only for participants censored before visit 3. Participants were followed from visit 1 through 31 December 2016 for incident diabetes. We used Cox regression models to characterize associations of aMed (quintiles as well as per 1-point higher) with incident diabetes adjusted for energy intake, age, sex, race and study center, and education (Model 1) for all participants then stratified by race and body mass index (BMI). Model 2 included potential mediating behavioral and clinical measures associated with diabetes. Results are presented as hazard ratios and 95% confidence intervals. Over a median follow-up of 22 years, there were 4024 incident cases of diabetes. Higher aMed scores were associated with lower diabetes risk [Model 1: 0.83 (0.73-0.94) for Q5 vs Q1 (p-trend < 0.001) and 0.96 (0.95-0.98) for 1-point higher]. Associations were stronger for black vs white participants (interaction p < 0.001) and weaker for obese vs normal BMI (interaction p < 0.01). Associations were attenuated but statistically significant in Model 2. An eating pattern high in fruits, vegetables, whole grains, legumes, nuts, and fish, and moderate in alcohol was associated with a lower risk of diabetes in a community-based U.S.


The development of vaccination perspectives among chiropractic, naturopathic and medical students: a case study of professional enculturation.
An important influence on parents' decisions about pediatric vaccination (children under 6 years of age) is the attitude of their health care providers, including complementary and alternative medicine (CAM) providers. Very limited qualitative research exists, however, on how attitudes towards vaccination develop among healthcare professionals in-training. We explored perspective development among three groups of students: medical, chiropractic, and naturopathic. We conducted focus group sessions with participants from each year of study at three different healthcare training programs in Ontario, Canada. Semi-structured and open-ended questions were used to elicit dynamic interaction among participants and explore how they constructed their attitudes toward vaccination at the beginning and part way through their professional training. Analyses of verbatim transcripts of audiotaped interviews were conducted both inductively and deductively using questions structured by existing literature on learning, professional socialization and interprofessional relations. We found five major themes and each theme was illustrated with representative quotes. Numerous unexpected insights emerged within these themes, including students' general open-mindedness towards pediatric vaccination at the beginning of their training; the powerful influence of both formal education and informal socialization; uncritical acceptance of the vaccination views of senior or respected professionals; students' preference for multiple perspectives rather than one-sided, didactic instruction; the absence of explicit socio-cultural tensions among professions; and how divergences among professional students' perspectives result from differing emphases with respect to lifestyle, individual choice, public health and epidemiological factors-rather than disagreement concerning the biomedical evidence. This last finding implies that their different perspectives on pediatric vaccination may be complementary rather than irreconcilable. Our findings should be considered by developers of professional and interprofessional educational curricula and public health officials formulating policy on pediatric vaccination.


Wortmannin inhibits pkb/akt phosphorylation and promotes gemcitabine antitumor activity in orthotopic human pancreatic cancer xenografts in immunodeficient mice.
Pancreatic cancer is resistant to almost all classes of cytotoxic agents. Gemcitabine seems to be the current drug of choice. We have recently reported that inhibition of the phosphatidylinositide 3-kinase-protein kinase B (PKB/Akt) cell survival pathway by wortmannin enhances gemcitabine-induced apoptosis in cultured human pancreatic cancer cells (1). The present study investigated the effects of wortmannin on orthotopic human pancreatic cancer xenografts implanted in severe combined immunodeficient mice. Animals were given single i.v. bolus injections of 0.175, 0.35, or 0.7 mg/kg of wortmannin and killed at 0.5, 1, 2, or 4 h after treatment. Phosphorylated PKB/Akt levels in tumor tissues were measured by fluorescence image analysis. Wortmannin was found to inhibit PKB/Akt phosphorylation in a time- and dose-dependent manner, reaching a plateau at 4 h and at 0.7 mg/kg. The levels of phosphorylated PKB/Akt were maximally decreased by approximately 50% relative to the vehicle control. Subsequently, the extent of apoptosis in tumors treated with gemcitabine or wortmannin alone or in combination was determined using terminal deoxynucleotidyl transferase-mediated nick end labeling assay and computerized image analysis. Orthotopic tumors exposed to 80 mg/kg gemcitabine for 48 h and then 0.7 mg/kg wortmannin for 4 h showed a 5-fold increase (P = 0.002) in apoptosis compared with those treated with each agent alone and with the vehicle control. The combination treatment also significantly (P < 0.001) inhibited tumor growth. Taken together, our findings support the potential of phosphatidylinositide 3-kinase inhibitors as adjuncts to conventional chemotherapy in the treatment of pancreatic cancer.


Self-speech-induced facilitation of simple reaching movements in persons with stroke.
Occupational therapists are routinely involved in upper-extremity rehabilitation in persons with stroke. Recently we have reported that self-speech can be used to facilitate simple reach in young and older adults. The purpose of this study is to examine whether self-speech-induced facilitation of simple reach can be translated in persons with stroke. Following a counterbalanced repeated-measure design, seven persons with right brain stroke performed reaching and placing a glass on a cabinet shelf under four concurrent speech conditions of vocalizing the word "yeah" with both hemiplegic and nonhemiplegic sides. These four conditions are self-vocalization, external vocalization, imaginary vocalization, and no vocalization. Consistent with our previous study, in this study we also recorded the movement kinematics of angular movement of the upper arm. Reaching velocity was significantly higher under self-vocalization conditions compared to the reaching velocity under no vocalization or other vocalization conditions (p < .05). Motor performance in terms of velocity, nonhemiparetic side was significantly better than hemiparetic side (p < .05). The results suggest that self-speech can be used in facilitating movements in patients with stroke. When working with patients with right hemispheric stroke, therapists might explore possibilities of using patient's self-speech to enhance the quality of movement performance.


New insights into the GABA(A) receptor structure and orthosteric ligand binding: receptor modeling guided by experimental data.
GABA(A) receptors (GABA(A)Rs) are ligand gated chloride ion channels that mediate overall inhibitory signaling in the CNS. A detailed understanding of their structure is important to gain insights in, e.g., ligand binding and functional properties of this pharmaceutically important target. Homology modeling is a necessary tool in this regard because experimentally determined structures are lacking. Here we present an exhaustive approach for creating a high quality model of the α(1)β(2)γ(2) subtype of the GABA(A)R ligand binding domain, and we demonstrate its usefulness in understanding details of orthosteric ligand binding. The model was constructed by using multiple templates and by incorporation of knowledge from biochemical/pharmacological experiments. It was validated on the basis of objective energy functions, its ability to account for available residue specific information, and its stability in molecular dynamics (MD) compared with that of the two homologous crystal structures. We then combined the model with extensive structure-activity relationships available from two homologous series of orthosteric GABA(A)R antagonists to create a detailed hypothesis for their binding modes. Excellent agreement with key experimental data was found, including the ability of the model to accommodate and explain a previously developed pharmacophore model. A coupling to agonist binding was thereby established and discussed in relation to activation mechanisms. Our results highlight the importance of critical evaluation and optimization of each step in the homology modeling process. The approach taken here can greatly aid in increasing the understanding of GABA(A)Rs and related receptors where structural insight is limited and reliable models are difficult to obtain.


Neonatal cardiac dysfunction and transcriptome changes caused by the absence of Celf1.
The RNA binding protein Celf1 regulates alternative splicing in the nucleus and mRNA stability and translation in the cytoplasm. Celf1 is strongly down-regulated during mouse postnatal heart development. Its re-induction in adults induced severe heart failure and reversion to fetal splicing and gene expression patterns. However, the impact of Celf1 depletion on cardiac transcriptional and posttranscriptional dynamics in neonates has not been addressed. We found that homozygous Celf1 knock-out neonates exhibited cardiac dysfunction not observed in older homozygous animals, although homozygous mice are smaller than wild type littermates throughout development. RNA-sequencing of mRNA from homozygous neonatal hearts identified a network of cell cycle genes significantly up-regulated and down-regulation of ion transport and circadian genes. Cell cycle genes are enriched for Celf1 binding sites supporting a regulatory role in mRNA stability of these transcripts. We also identified a cardiac splicing network coordinated by Celf1 depletion. Target events contain multiple Celf1 binding sites and enrichment in GU-rich motifs. Identification of direct Celf1 targets will advance our knowledge in the mechanisms behind developmental networks regulated by Celf1 and diseases where Celf1 is mis-regulated.


Etomidate: to use or not to use for endotracheal intubation in the critically ill?
Endotracheal intubation is frequently performed in the intensive care unit (ICU). It can be life-saving for many patients who present with acute respiratory distress. However, it is equally associated with complications that may lead to unwanted effects in this patient population. According to the literature, the rate of complications associated with endotracheal intubation is much higher in an environment such as the ICU as compared to other, more controlled environments (i.e., operating room). Thus, the conduct of performing such a procedure needs to be accomplished with the utmost care. To facilitate establishment of the breathing tube, sedation is routinely administered. Given the tenuous hemodynamic status of the critically ill, etomidate was frequently chosen to blunt further decreases in blood pressure and/or heart rate. Recently however, reports have demonstrated a possible association with the use of etomidate for endotracheal intubation and mortality in the critically ill. In addition, this association seems to be predominantly in patients diagnosed with sepsis. As a result, some have advocated against the use of this medication in septic patients. Due to the negative associations identified with etomidate and mortality, several investigators have evaluated potential alternatives to this solution (e.g., ketamine and ketamine-propofol admixture). These studies have shown promise. However, despite the evidence against using etomidate for endotracheal intubation, other studies have demonstrated no such association. This leaves the critical care clinician with uncertainty regarding the best sedative to administer in this patient population. The following editorial discusses current evidence regarding etomidate use for endotracheal intubation and mortality. In particular, we highlight a recent article with the largest population to date that found no association between etomidate and mortality in the critically ill and illustrate important findings that the reader should be aware of regarding this article.


Pharmacokinetics of antiretroviral regimens containing tenofovir disoproxil fumarate and atazanavir-ritonavir in adolescents and young adults with human immunodeficiency virus infection.
The primary objective of this study was to measure atazanavir-ritonavir and tenofovir pharmacokinetics when the drugs were used in combination in young adults with human immunodeficiency virus (HIV). HIV-infected subjects > or =18 to <25 years old receiving (> or =28 days) 300/100 mg atazanavir-ritonavir plus 300 mg tenofovir disoproxil fumarate (TDF) plus one or more other nucleoside analogs underwent intensive 24-h pharmacokinetic studies following a light meal. Peripheral blood mononuclear cells were obtained at 1, 4, and 24 h postdose for quantification of intracellular tenofovir diphosphate (TFV-DP) concentrations. Twenty-two subjects were eligible for analyses. The geometric mean (95% confidence interval [CI]) atazanavir area under the concentration-time curve from 0 to 24 h (AUC(0-24)), maximum concentration of drug in serum (C(max)), concentration at 24 h postdose (C(24)), and total apparent oral clearance (CL/F) values were 35,971 ng x hr/ml (30,853 to 41,898), 3,504 ng/ml (2,978 to 4,105), 578 ng/ml (474 to 704), and 8.3 liter/hr (7.2 to 9.7), respectively. The geometric mean (95% CI) tenofovir AUC(0-24), C(max), C(24), and CL/F values were 2,762 ng.hr/ml (2,392 to 3,041), 254 ng/ml (221 to 292), 60 ng/ml (52 to 68), and 49.2 liter/hr (43.8 to 55.3), respectively. Body weight was significantly predictive of CL/F for all three drugs. For every 10-kg increase in weight, there was a 10%, 14.8%, and 6.8% increase in the atazanavir, ritonavir, and tenofovir CL/F, respectively (P < or = 0.01). Renal function was predictive of tenofovir CL/F. For every 10 ml/min increase in creatinine clearance, there was a 4.6% increase in tenofovir CL/F (P < 0.0001). The geometric mean (95% CI) TFV-DP concentrations at 1, 4, and 24 h postdose were 96.4 (71.5 to 130), 93.3 (68 to 130), and 92.7 (70 to 123) fmol/million cells. There was an association between renal function, tenofovir AUC, and tenofovir C(max) and intracellular TFV-DP concentrations, although none of these associations reached statistical significance. In these HIV-infected young adults treated with atazanavir-ritonavir plus TDF, the atazanavir AUC was similar to those of older adults treated with the combination. Based on data for healthy volunteers, a higher tenofovir AUC may have been expected, but was not seen in these subjects. This might be due to faster tenofovir CL/F because of higher creatinine clearance in this age group. Additional studies of the exposure-response relationships of this regimen in children, adolescents, and adults would advance our knowledge of its pharmacodynamic properties.


Long range surface plasmon-coupled fluorescence emission for biosensor applications.
A biosensor scheme that employs long range surface plasmons (LRSPs) for the efficient excitation and collection of fluorescence light from fluorophore-labeled biomolecules captured in a three-dimensional hydrogel matrix is discussed. This new approach to plasmon-enhanced fluorescence (PEF) is experimentally and theoretically investigated by using the Kretschmann configuration of attenuated total reflection (ATR) method. A layer structure supporting LRSPs that consists of a low refractive index fluoropolymer layer, a thin gold film and a large binding capacity N-isopropylacrylamide (NIPAAm)-based hydrogel matrix swollen in an aqueous sample is employed. By using this layer architecture, the extended field of LRSPs probes the binding of biomolecules in the binding matrix at up to micrometer distances from the gold surface. With respect to regular surface plasmon-enhanced fluorescence spectroscopy (SPFS) and surface plasmon-coupled emission (SPCE), a narrower angular distribution of the fluorescence light intensity, a larger peak intensity and the excitation and emission at lower angles were observed.


Seroprevalence of Hepatitis E Virus in Iran: A Systematic Review and Meta-analysis.
BACKGROUND Hepatitis E virus (HEV) is one of common causes of viral hepatitis worldwide with higher prevalence in tropical and subtropical regions. Although epidemics of HEV have been reported from Iran, there are variable reports of this infection out of epidemics from different parts of Iran. This study aimed to determine the seroprevalence of HEV in Iran. METHODS In this systematic review and meta-analysis we searched PubMed, Scopus, Science direct, Google Scholar, Scientific Information Databank (SID), IranMedex, and Magiran for all relevant studies published in either English or Persian languages, up to 2015. Pooled seroprevalence estimates with a DerSimonian-Laird random-effects model were calculated. Statistical heterogeneity among the included studies was evaluated by Cochrane Q statistic and I2. RESULTS 38 studies fulfilled the inclusion criteria compromising 18461 participants. The pooled seroprevalence rate of HEV in Iran was estimated about 10% (95% CI=0.09-0.12) with maximum and minimum of 46% (95 % CI=0.42-0.50), and 0.01% (95 % CI=0.000-0.002), respectively. CONCLUSION HEV is common in Iran although the prevalence is lower than some neighbor countries.


Detection and visualization of abnormality in chest radiographs using modality-specific convolutional neural network ensembles.
Convolutional neural networks (CNNs) trained on natural images are extremely successful in image classification and localization due to superior automated feature extraction capability. In extending their use to biomedical recognition tasks, it is important to note that visual features of medical images tend to be uniquely different than natural images. There are advantages offered through training these networks on large scale medical common modality image collections pertaining to the recognition task. Further, improved generalization in transferring knowledge across similar tasks is possible when the models are trained to learn modality-specific features and then suitably repurposed for the target task. In this study, we propose modality-specific ensemble learning toward improving abnormality detection in chest X-rays (CXRs). CNN models are trained on a large-scale CXR collection to learn modality-specific features and then repurposed for detecting and localizing abnormalities. Model predictions are combined using different ensemble strategies toward reducing prediction variance and sensitivity to the training data while improving overall performance and generalization. Class-selective relevance mapping (CRM) is used to visualize the learned behavior of the individual models and their ensembles. It localizes discriminative regions of interest (ROIs) showing abnormal regions and offers an improved explanation of model predictions. It was observed that the model ensembles demonstrate superior localization performance in terms of Intersection of Union (IoU) and mean Average Precision (mAP) metrics than any individual constituent model.


Safety evaluation of the food enzyme α-glucosidase from the Aspergillus niger strain AE-TGU.
The food enzyme α-glucosidase (α-d-glucoside glucohydrolase; EC 18.104.22.168) is produced with the non-genetically modified Aspergillus niger strain AE-TGU by Amano Enzyme Inc. The food enzyme is free from viable cells of the production organism. The food enzyme is intended to be used in baking processes, cereal-based processes, brewing processes and starch processing for the production of glucose syrups and other starch hydrolysates. Since residual amounts of total organic solids (TOS) are removed by the purification steps applied during the production of glucose syrups, dietary exposure was only calculated for the remaining three food processes. Based on the maximum use levels recommended, dietary exposure was estimated to be up to 0.64 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 1,062 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, results in a margin of exposure of at least 1,650. A search for similarity of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood for this to occur is considered to be low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.


Detection of glycomic alterations induced by overexpression of p-glycoprotein on the surfaces of L1210 cells using sialic acid binding lectins.
P-glycoprotein (P-gp) overexpression is the most frequently observed cause of multidrug resistance in neoplastic cells. In our experiments, P-gp was expressed in L1210 mice leukemia cells (S cells) by selection with vincristine (R cells) or transfection with the gene encoding human P-gp (T cells). Remodeling of cell surface sugars is associated with P-gp expression in L1210 cells as a secondary cellular response. In this study, we monitored the alteration of cell surface saccharides by Sambucus nigra agglutinin (SNA), wheat germ agglutinin (WGA) and Maackia amurensis agglutinin (MAA). Sialic acid is predominantly linked to the surface of S, R and T cells via α-2,6 branched sugars that tightly bind SNA. The presence of sialic acid linked to the cell surface via α-2,3 branched sugars was negligible, and the binding of MAA (recognizing this branch) was much less pronounced than SNA. WGA induced greater cell death than SNA, which was bound to the cell surface and agglutinated all three L1210 cell-variants more effectively than WGA. Thus, the ability of lectins to induce cell death did not correlate with their binding efficiency and agglutination potency. Compared to S cells, P-gp positive R and T cells contain a higher amount of N-acetyl-glucosamine on their cell surface, which is associated with improved WGA binding. Both P-gp positive variants of L1210 cells are strongly resistant to vincristine as P-gp prototypical drug. This resistance could not be altered by liberalization of terminal sialyl residues from the cell surface by sialidase.


Semiblind Spectral Factorization Approach for Magnetic Resonance Spectroscopy Quantification.
An observed magnetic resonance (MR) spectrum is composed of a set of metabolites spectrum, baseline, and noise. Quantification of metabolites of interest in the MR spectrum provides great opportunity for early diagnosis of dangerous disease such as brain tumors. In this paper, a novel spectral factorization approach based on singular spectrum analysis (SSA) is proposed to quantify magnetic resonance spectroscopy (MRS). In addition, baseline removal is performed in this study. The proposed method is a semiblind spectral factorization algorithm that jointly uses observed signal and prior knowledge about metabolites of interest to improve metabolite separation. In order to incorporate prior knowledge about metabolites of interest, a new covariance matrix is suggested that exploits correlation between the observed nuclear magnetic resonance signal and prior knowledge. The objectives of the proposed method are 1) removing baseline in frequency domain using SSA; 2) extracting the underlying components of MRS signal based on the suggested novel covariance matrix; and 3) reconstructing metabolite of interest by combining some of the extracted components using a novel cost function. Performance of the proposed method is evaluated using both synthetic and real MRS signals. The obtained results show the effectiveness of the proposed technique to accurately remove baseline and extract metabolites of MRS signal.


4 mm long vs longer implants in augmented bone in posterior atrophic jaws: 1-year post-loading results from a multicentre randomised controlled trial.
To evaluate whether 4.0 mm short dental implants could be an alternative to augmentation with xenographs in the maxilla and placement of at least 10.0 mm long implants in posterior atrophic jaws. A group of 40 patients with atrophic posterior (premolar and molar areas) mandibles with 5.0 mm to 6.0 mm bone height above the mandibular canal and 40 patients with atrophic maxillas having 4.0 mm to 5.0 mm below the maxillary sinus, were randomised according to a parallel group design to receive between one and three 4.0 mm long implants or one to three implants of at least 10.0 mm long in augmented bone, at two centres. All implants had a diameter of 4.0 mm or 4.5 mm. Mandibles were vertically augmented with inter-positional equine bone blocks and resorbable barriers. Implants were placed 4 months after the inter-positional grafting. Maxillary sinuses were augmented with particulated porcine bone via a lateral window covered with resorbable barriers, and implants were placed simultaneously. Implants were not submerged and were loaded after 4 months with provisional screw-retained reinforced acrylic restorations replaced after another 4 months by definitive screw-retained metal-composite prostheses. Patients were followed up to 1 year post-loading. Outcome measures were: prosthesis and implant failures, any complication, and peri-implant marginal bone level changes. Three patients dropped out; one from the maxillary augmented group, one from the mandibular augmented group, and one from the maxillary short implant group. In six augmented mandibles (30%) it was not possible to place implants of at least 10.0 mm, so shorter implants were placed instead. In mandibles, one implant from the augmented group failed vs two 4.0 mm implants in two patients from the short implant group. In maxillae, three short implants failed in two patients vs seven long implants in four patients (two long implants and one short implant dropped into the maxillary sinus). Two prostheses on short implants (one mandibular and one maxillary) were placed at a later stage because of implant failures, vs six prostheses (one mandibular and five maxillary) at augmented sites (one mandibular prosthesis not delivered, three maxillary prostheses delivered with delays, one not delivered, and one failed) at augmented sites. In particular, three patients in the augmented group (one mandible and two maxillae) were not wearing a prosthesis. There were no statistically significant differences in implant failures (P (chi-square test) = 0.693; difference in proportion = 0.03; CI 95% -0.11 to 0.17) or prostheses failures (P (chi-square test) = 0.126; difference in proportion = 0.10; CI 95% -0.03 to 0.24). At mandibular sites, nine augmented patients were affected by complications vs two patients treated with short implants (P (chi-square test) = 0.01; difference in proportion = 0.37; CI 95% 0.11 to 0.63), the difference being statistically significant. No significant differences were found for maxillae: nine sinus-lifted patients vs four short implant patients were affected by complications (P (chi-square test) = 0.091; difference in proportion = 0.25; CI 95% -0.03 to 0.53). At 1-year post-loading, average peri-implant bone loss was 0.51 mm at 4 mm long mandibular implants, 0.77 mm at 10 mm or longer mandibular implants, 0.63 mm at short maxillary implants and 0.72 mm at long maxillary implants. The difference was statistically significant in mandibles (mean difference -0.26 mm, 95% CI -0.39 to -0.13, P (ANCOVA) < 0.001), but not in maxillae (mean difference -0.09 mm, 95% CI -0.24 to 0.05, P (ANCOVA) = 0.196). One year after loading 4.0 mm long implants achieved similar results, if not better, than longer implants in augmented jaws, but were affected by fewer complications. Short implants might be a preferable choice over bone augmentation, especially in mandibles, since the treatment is less invasive, faster, cheaper, and associated with less morbidity. However, 5 to 10 years post-loading data are necessary before making reliable recommendations.