Capsular serotype-specific attack rates and duration of carriage of Streptococcus pneumoniae in a population of children.
The relative invasiveness rates (attack rates) of Streptococcus pneumoniae of different capsular serotypes in children are not known. Estimates of capsular serotype invasiveness (designated "invasive odds ratios") that are based on cross-sectional prevalence carriage data have been published, but these estimates could be biased by variation in the duration of carriage. The relative attack rates of invasive pneumococci were measured using national UK surveillance data on invasive pneumococcal disease (IPD) incidence and data on incidence of pneumococcal acquisition from longitudinal studies of nasopharyngeal pneumococcal carriage. We found significant differences in capsular serotype-specific attack rates. For example, capsular serotypes 4, 14, 7F, 9V, and 18C were associated with rates of >20 IPD cases/100,000 acquisitions, whereas capsular serotypes 23F, 6A, 19F, 16F, 6B, and 15B/C were associated with <10 IPD cases/100,000 acquisitions. There was an inverse relationship between duration of carriage and attack rate by capsular serotype (P<.0001). Attack rates were significantly correlated with invasive odds ratios (P<.0001). The capsular serotype is a major determinant of both pneumococcal duration of carriage and attack rate. Published invasive odds ratios are a reliable and practical method of determining capsular serotype invasiveness and will be valuable for investigating and characterizing emerging capsular serotypes in the context of conjugate vaccination.


Rac1 augments Wnt signaling by stimulating β-catenin-lymphoid enhancer factor-1 complex assembly independent of β-catenin nuclear import.
β-Catenin transduces the Wnt signaling pathway and its nuclear accumulation leads to gene transactivation and cancer. Rac1 GTPase is known to stimulate β-catenin-dependent transcription of Wnt target genes and we confirmed this activity. Here we tested the recent hypothesis that Rac1 augments Wnt signaling by enhancing β-catenin nuclear import; however, we found that silencing/inhibition or up-regulation of Rac1 had no influence on nuclear accumulation of β-catenin. To better define the role of Rac1, we employed proximity ligation assays (PLA) and discovered that a significant pool of Rac1-β-catenin protein complexes redistribute from the plasma membrane to the nucleus upon Wnt or Rac1 activation. More importantly, active Rac1 was shown to stimulate the formation of nuclear β-catenin-lymphoid enhancer factor 1 (LEF-1) complexes. This regulation required Rac1-dependent phosphorylation of β-catenin at specific serines, which when mutated (S191A and S605A) reduced β-catenin binding to LEF-1 by up to 50%, as revealed by PLA and immunoprecipitation experiments. We propose that Rac1-mediated phosphorylation of β-catenin stimulates Wnt-dependent gene transactivation by enhancing β-catenin-LEF-1 complex assembly, providing new insight into the mechanism of cross-talk between Rac1 and canonical Wnt/β-catenin signaling.


RNA polymerase II termination involves C-terminal-domain tyrosine dephosphorylation by CPF subunit Glc7.
At the 3' ends of protein-coding genes, RNA polymerase (Pol) II is dephosphorylated at tyrosine residues (Tyr1) of its C-terminal domain (CTD). In addition, the associated cleavage-and-polyadenylation factor (CPF) cleaves the transcript and adds a poly(a) tail. Whether these events are coordinated and how they lead to transcription termination remains poorly understood. Here we show that CPF from Saccharomyces cerevisiae is a Pol II-CTD phosphatase and that the CPF subunit Glc7 dephosphorylates Tyr1 in vitro. In vivo, the activity of Glc7 is required for normal Tyr1 dephosphorylation at the polyadenylation site, for recruitment of termination factors Pcf11 and Rtt103 and for normal Pol II termination. These results show that transcription termination involves Tyr1 dephosphorylation of the CTD and indicate that pre-mRNA processing by CPF and transcription termination are coupled via Glc7-dependent Pol II-Tyr1 dephosphorylation.


Rapid Nanogram Scale Screening Method of Microarrays to Evaluate Drug-Polymer Blends Using High-Throughput Printing Technology.
A miniaturized, high-throughput assay was optimized to screen polymer-drug solid dispersions using a 2-D Inkjet printer. By simply printing nanoliter amounts of polymer and drug solutions onto an inert surface, drug/polymer microdots of tunable composition were produced in an easily addressable microarray format. The amount of material printed for each dried spot ranged from 25 ng to 650 ng. These arrays were used to assess the stability of drug/polymer dispersions with respect to recrystallization, using polarized light microscopy. One array with a panel of 6 drugs formulated at different ratios with a poly(vinylpyrrolidone-vinyl acetate) (PVPVA) copolymer was developed to estimate a possible bulk (gram-scale) approximation threshold from the final printed nanoamount of formulation. Another array was printed at a fixed final amount of material to establish a literature comparison of one drug formulated with different commercial polymers for validation. This new approach may offer significant efficiency in pharmaceutical formulation screening, with each experiment in the nanomicro-array format requiring from 3 up to 6 orders of magnitude lower amounts of sample than conventional screening methods.


Venous access using the bovine carotid heterograft: techniques, results, and complications in 75 patients.
Between January 1974 and March 1977, arm bovine carotid heterograft arteriovenous fistulas were constructed in 75 patients. Twenty-six fistulas were established between the distal radial artery and an antecubital vein, and 49 fistulas used the brachial artery and the axillary vein. Thirty-eight heterografts have functioned without complication over a maximun interval of 29 months, including 15 of 26 radiobasilic fistulas and 23 of 49 brachioaxillary fistulas. Of the 37 heterograft failures, 28 were caused by thrombosis, six by infection, two by false aneurysm formation, and one by distal arm ischemia. Early postoperative thrombosis was associated with a higher subsequent failure rate after thrombectomy than was late thrombosis and it often required insertion of a new heterograft. Infected heterografts must be ligated and eventually replaced with another fistula at a distant site to avoid the potentially lethal complications of systemic sepsis and local hemorrhage.


Estimation of beverage consumption and associated caloric intake in adult Czech population. An observational study.
Food intake is a commonly monitored issue in many studies. In contrast, almost no information has been published on beverage intake in adults. To evaluate beverage intake, we studied a population of 1, 200 adults (656 males and 544 females, aged 18-54 years). The volumes and types of beverages were obtained from self-reported questionnaires. The mean beverage intake was highly variable, with a minimum of 450 mL/day and a maximum of 5,330 mL/day. A mean of 1,575 mL/day was found in the entire population (2,300 mL in males and 840 mL in females). Different patterns in the consumption of beverage types were observed between the males and females. For both males and females, the most common beverage consumed was water followed by tea. The next preferable beverages were alcoholic beer, coffee, and non-alcoholic beer in males and coffee, milk, and alcoholic beer in females. The estimated caloric intake from beverages covers, in most individuals, 10-30% of the recommended daily caloric intake. There is substantial variation among individuals, both in beverage intake and in caloric intake through beverages. The caloric intake from beverages reaches, in some individuals, one-third of the recommended daily caloric rate.


Heme protein-induced chronic renal inflammation: suppressive effect of induced heme oxygenase-1.
Heme oxygenase (HO) is the rate-limiting enzyme in the degradation of heme; its inducible isozyme, HO-1, protects against acute heme protein-induced nephrotoxicity and other forms of acute tissue injury. This study examines the induction of HO-1 in the kidney chronically inflamed by heme proteins and the functional significance of such an induction of HO-1. Studies were undertaken in a patient with chronic tubulointerstitial disease in the setting of paroxysmal nocturnal hemoglobinuria (PNH), in a rat model of chronic tubulointerstitial nephropathy caused by repetitive exposure to heme proteins, and in genetically engineered mice deficient in HO-1 (HO-1 -/-) in which hemoglobin was repetitively administered. The kidney in PNH evinces robust induction of HO-1 in renal tubules in the setting of chronic inflammation. The heme protein-enriched urine from this patient, but not urine from a healthy control subject, induced expression of HO-1 in renal tubular epithelial cells (LLC-PK1 cells). A similar induction of HO-1 and related findings are recapitulated in a rat model of chronic inflammation induced by repetitive exposure to heme proteins. Additionally, in the rat, the administration of heme proteins induces monocyte chemoattractant protein (MCP-1). The functional significance of HO-1 so induced was uncovered in the HO-1 knockout mouse: Repeated administration of hemoglobin to HO-1 +/+ and HO-1 -/- mice led to intense interstitial cellular inflammation in HO-1 -/- mice accompanied by striking up-regulation of MCP-1 and activation of one of its stimulators, nuclear factor-kappaB (NF-kappaB). These findings were not observed in similarly treated HO-1 +/+ mice or in vehicle-treated HO-1 -/- and HO-1 +/+ mice. We conclude that up-regulation of HO-1 occurs in the kidney in humans and rats repetitively exposed to heme proteins. Such up-regulation represents an anti-inflammatory response since the genetic deficiency of HO-1 markedly increases activation of NF-kappaB, MCP-1 expression, and tubulointerstitial cellular inflammation.


Transplant Center Volume and the Risk of Pancreas Allograft Failure.
Successful pancreas transplantation requires surgical expertise and multidisciplinary medical management. The impact of transplant center volume on pancreas allograft survival remains unclear. We examined Organ Procurement and Transplantation Network data on 11 568 simultaneous pancreas-kidney (SPK) and 4308 solitary pancreas (pancreas transplant alone and pancreas after kidney) transplants between 2000 and 2013. Average annual transplant center volume was categorized by tertiles into low, medium, and high volume, respectively, as follows: 1 to 6 (n = 3861), 7 to 13 (n = 3891), and 14 to 34 (n = 3888) for SPK, and 1 to 3 (n = 1417), 4 to 10 (n = 1518), and 11 to 33 (n = 1377) for solitary pancreas transplants. Favorable donor characteristics were seen in low-volume centers. For SPK transplantation, low (adjusted hazard ration [aHR], 1.55, 95% confidence interval [CI], 1.34-1.8) and medium (aHR, 1.24; 95% CI, 1.07-1.44) center volumes were associated with a higher risk of early pancreas graft failure at 3 months. The increased risk associated with low center volume extended to 1, 5, and 10 years. For solitary pancreas transplants, low, but not medium, center volume was associated with a higher risk of early pancreas graft failure at 3 months (aHR, 1.56; 95% CI, 1.232-1.976), and this risk persisted over 10 years. Patients transplanted at high-volume centers had better pancreas survival rates across all categories of the Pancreas Donor Risk Index. On average, low center volume were associated with higher risk for pancreas failure. Future studies should seek to identify care processes that support optimal outcomes after pancreas transplantation irrespective of center volume.


Quantitative evaluation of factors influencing the repeatability of SD-OCT thickness measurements in the rat.
To quantify repeatability and reproducibility of thickness measurements and the effects of realignment and image quality on measurements of retinal thickness from optical coherence tomographic (OCT) imaging in the rat eye. Retinal imaging was performed in 16 brown norway rats (N = 16 EYES; X = 372 G). Precision metrics: 95% limits of agreement (LoA), intraclass correlation coefficient (ICC), and the coefficient of variation (CV), were calculated using manual and combined manual + automated realignment procedures for nerve fiber and retinal ganglion cell layer (NFL/GCL), NFL/GCL and inner plexiform layer (NFL/GCL + IPL), and total retina thicknesses (excluding blood vessels). The influence of image quality on NFL thickness measurement was assessed by comparing high- and low-quality image data (real and simulated) from the rat as well as clinical data. Mean NFL/GCL thickness was 26 3 M, NFL/GCL + IPL thickness was 70 3 M, AND total retinal thickness was 192 7 M. Thickness difference between imaging sessions for NFL/GCL was 1 M (95% LOA: -4 to 3 μm; ICC = 0.82; CV = 4.7%), for NFL/GCL + IPL was 0 μm (95% LoA: -4 to 4 μm; ICC = 0.88; CV = 1.4%), and total retinal thickness was 1 μm (95% LoA: -3 to 4 μm; ICC = 0.97; CV = 0.7%). Thickness differences were similar between realignment procedures (NFL/GCL: P = 0.43; NFL/GCL + IPL: P = 0.33; total retina: P = 0.62). Although NFL thickness measurements increased slightly in low-quality rat images (4 μm; P = 0.04), this was not true with clinical images (1.4 μm; P = 0.36). Precision of retinal layer thickness estimation from OCT imaging is excellent when manual and automated realignment procedures are combined, but may still be influenced by image quality and segmentation methods.


Unsupervised vector-based classification of single-molecule charge transport data.
The stochastic nature of single-molecule charge transport measurements requires collection of large data sets to capture the full complexity of a molecular system. Data analysis is then guided by certain expectations, for example, a plateau feature in the tunnelling current distance trace, and the molecular conductance extracted from suitable histogram analysis. However, differences in molecular conformation or electrode contact geometry, the number of molecules in the junction or dynamic effects may lead to very different molecular signatures. Since their manifestation is a priori unknown, an unsupervised classification algorithm, making no prior assumptions regarding the data is clearly desirable. Here we present such an approach based on multivariate pattern analysis and apply it to simulated and experimental single-molecule charge transport data. We demonstrate how different event shapes are clearly separated using this algorithm and how statistics about different event classes can be extracted, when conventional methods of analysis fail.


Safe-water shortages, gender perspectives, and related challenges in developing countries: the case of Uganda.
The need for water continues to become more acute with the changing requirements of an expanding world population. Using a logistical analysis of data from 301 respondents from households that harvest rainwater in Uganda, the relationship between dependent variables, such as water management performed as female-dominated practices, and independent variables, such as years of water harvesting, family size, tank operation and maintenance, and the presence of local associations, was investigated. The number of years of water harvesting, family size, tank operation and maintenance, and presence of local associations were statistically significantly related to adequate efficient water management. The number of years of water harvesting was linked to women's participation in household chores more than to the participation of men, the way of livelihoods lived for many years. Large families were concurrent with a reduction in water shortages, partially because of the availability of active labour. The findings also reveal important information regarding water-related operations and maintenance at the household level and the presence of local associations that could contribute some of the information necessary to minimise water-related health risks. Overall, this investigation revealed important observations about the water management carried out by women with respect to underlying safe-water shortages, gender perspectives, and related challenges in Uganda that can be of great importance to developing countries.


Mechanism of replication blocking and bypass of Y-family polymerase {eta} by bulky acetylaminofluorene DNA adducts.
Heterocyclic aromatic amines produce bulky C8 guanine lesions in vivo, which interfere and disrupt DNA and RNA synthesis. These lesions are consequently strong replication blocks. In addition bulky adducts give rise to point and frameshift mutations. The translesion synthesis (TLS) DNA polymerase η is able to bypass slowly C8 bulky adduct lesions such as the widely studied 2-aminofluorene-dG and its acetylated analogue mainly in an error-free manner. Replicative polymerases are in contrast fully blocked by the acetylated lesion. Here, we show that TLS efficiency of Pol η depends critically on the size of the bulky adduct forming the lesion. Based on the crystal structure, we show why the bypass reaction is so difficult and we provide a model for the bypass reaction. In our model, TLS is accomplished without rotation of the lesion into the anti conformation as previously thought.


Effects of Bretschneider cardioplegic fluid on the lysosomal cathepsins D and L of myocardium of coronary patients during coronary-aortal bypass graft operation.
We evaluated the effects of replacing the standard St. Thomas Hospital (ST) cardioplegic fluid with Bretschneider (BR) fluid during coronary-aortal bypass graft operation (CABG) in patients with coronary disease. BR fluid contains enormous concentration of histidine (approximately 0.2 mole L-1): thus increased levels of histamine or other metabolites might exert some cardiovascular effects. Therefore, the changes in the level of two lysosomal proteases: cathepsin D and L of myocardium were evaluated. Small fragments of right atrium (SFRA) were taken out during CABG: just before the injection of cold cardioplegia (control), at the peak of ischaemia and after reperfusion. Cathepsin D and L were assayed in SFRA homogenates at pH 3.6 (with haemoglobin as a substrate and pepstatin as cathepsin D inhibitor). The values of parameters Ai and Ae: being the indexes of intralysosomal (Ai) and extralysosomal (Ae) activation or inactivation of cathepsins were calculated. With BR cardioplegia--unlikely ST fluid--there is a significant increase in total activity (T) of both cathepsins but free (F) only for cathepsin L during ischaemia with come back (to the control level) during reperfusion period. T for both cathepsins and F for cathepsin L with BR are much greater than those with ST fluid both in ischaemia and reperfusion period. BR cardioplegia results in distinct, extralysosomal activation of cathepsin L during ischaemia instead of the inhibition found with ST fluid.


Asymmetric syntheses of methyl N,O-diacetyl-D-3-epi-daunosaminide and methyl N,O-diacetyl-D-ristosaminide.
Ab initio asymmetric syntheses of methyl N,O-diacetyl-D-3-epi-daunosaminide and methyl N,O-diacetyl-D-ristosaminide, employing diastereoselective epoxidation and dihydroxylation, respectively, of alkyl (3S,αR,Z)-3-[N-benzyl-N-(α-methylbenzyl)amino]hex-4-enoates as the key steps, are reported. The requisite substrates were readily prepared using the conjugate additions of lithium (R)-N-benzyl-N-(α-methylbenzyl)amide to methyl and tert-butyl (E)-hexa-2-en-4-ynoates followed by diastereoselective alkyne reduction. syn-Dihydroxylation using OsO4 proceeded under steric control on the 4Re,5Re face of the olefin to give the corresponding diol, which subsequently underwent lactonization. Meanwhile, epoxidation using F3CCO3H in conjunction with F3CCO2H proceeded on the opposite 4Si,5Si face of the olefin under hydrogen-bonding control from the in situ formed ammonium ion. Treatment of the intermediate epoxide with concd aq H2SO4 promoted highly regioselective ring-opening (distal to the in situ formed ammonium moiety) to give the corresponding diol (completing overall the formal anti-dihydroxylation of the olefin), which then underwent lactonization under the reaction conditions. Elaboration of these diastereoisomeric lactones through hydrogenolysis, N-Boc protection, reduction, methanolysis, and acetate protection gave methyl N,O-diacetyl-D-3-epi-daunosaminide and methyl N,O-diacetyl-D-ristosaminide.


Understanding the costs of support transactions in daily life.
The benefits of close relationships for mental and physical health are well documented. One of the mechanisms presumed to underlie these effects is social support, whereby close others provide practical and emotional assistance in times of need. Although there is no doubt that generalized perceptions of support availability are beneficial, research examining actual instances of support receipt has found unexpectedly mixed results. Receiving support sometimes has positive effects, but null or even negative effects are common. In this article, we review our multimethod program of research that seeks to understand and explain the costs of receiving social support. We focus on reductions in the recipient's sense of relationship equity and self-efficacy as mechanisms of this effect and examine a number of other moderating factors. Although we have found that receiving support incurs costs on average, there is considerable variability yet to be explained. Using diary data from 312 persons preparing to take a challenging exam, we examined the potential of individual differences in neuroticism, agreeableness, and attachment insecurity to explain variability in experienced support costs. We close with new questions about why received support may be beneficial or benign in some situations while being especially toxic in others.


Association of Autoimmune Hepatitis and Celiac Disease: Role of Gluten-Free Diet in Reversing Liver Dysfunction.
Autoimmune hepatitis (AIH) is a chronic inflammation of liver with unclear etiology. It is frequently associated other autoimmune diseases, and its association with celiac disease (CD) is well established. In this article, we describe the case of a 50-year-old male with long-standing AIH taking azathioprine for 10 years, evaluated for flares in transaminases. Despite adding high-dose corticosteroids, his transaminases and bilirubin remained high. Serology for CD was ordered, which revealed elevated tissue transglutaminase antibody IgG and endomysial IgA, which was further confirmed by endoscopic biopsy. Strict gluten-free diet was advised and now for over 2 years he is in remission with azathioprine and budesonide. This emphasizes the role of gluten-free diet in reversing liver dysfunction in patients with AIH, and clinicians should consider screening for CD in patients with AIH with persistent elevation of liver enzymes despite immunosuppressant treatment.


Kinematics of the asal rift (djibouti) determined from the deformation of fieale volcano.
Because of its subaerial exposure the Asal rift segment provides an exceptional opportunity to quantify the deformation field of an active rift and assess the contribution of tectonics and volcanism to rifting processes. The present topography of the Asal rift results from the tectonic dismemberment during the last 100,000 years of a large central volcanic edifice that formed astride the rift zone 300,000 to 100,000 years ago. Three-dimensional deformation of this volcano has been quantified from the combined analysis of the topography and geology. The analysis indicates that spreading at 17 to 29 millimeters per year in a N40 degrees +/- 5 degrees E direction accounts for most of the separation between Arabia and Somalia. The small topographic subsidence relative to extension suggests that tectonic thinning of the crust has been balanced by injection and underplating of magmatic material of near crustal density. The methodology developed in this study could also be applied to quantify deformation in relatively inaccessible areas where the main available information is topography or bathymetry.


Long-term results of lower energy transurethral microwave thermotherapy.
We evaluate long-term results of lower energy transurethral microwave thermotherapy (Prostasoft 2.0*) and identify pretreatment characteristics that predict a favorable outcome. Between December 1990 and December 1992, 231 patients with lower urinary tract symptoms were treated with lower energy transurethral microwave thermotherapy. Subjective and objective voiding parameters were collected from medical records and a self-administered questionnaire. Kaplan-Meier plots were constructed to assess the risk of re-treatment. Of the patients 41% underwent invasive re-treatment within 5 years of followup and 17% were re-treated with medication. The re-treatment-free period was somewhat longer in patients with a peak flow rate greater than 10 ml. per second, a Madsen score 15 or less, a post-void residual volume 100 ml. or less and age greater than 65 years at baseline. Prostate volume did not modify the outcome. No incontinence was caused by transurethral microwave thermotherapy, 8% had recurrent urinary tract infection and 8% had retrograde ejaculation. Only 1 patient had a urethral stricture after transurethral microwave thermotherapy. At 5 years after transurethral microwave thermotherapy 41% of the patients received instrumental treatment. Patients with a lower Madsen score and lower residual volume, and those with higher peak flow and age were somewhat better responders to lower energy transurethral microwave thermotherapy.


Cognitive decline and dementia in the oldest-old.
The oldest-old are the fastest growing segment of the Western population. Over half of the oldest-old will have dementia, but the etiology is yet unknown. Age is the only risk factor consistently associated with dementia in the oldest-old. Many of the risk and protective factors for dementia in the young elderly, such as ApoE genotype, physical activity, and healthy lifestyle, are not relevant for the oldest-old. Neuropathology is abundant in the oldest-old brains, but specific pathologies of Alzheimer's disease (AD) or vascular dementia are not necessarily correlated with cognition, as in younger persons. It has been suggested that accumulation of both AD-like and vascular pathologies, loss of synaptic proteins, and neuronal loss contribute to the cognitive decline observed in the oldest-old. Several characteristics of the oldest-old may confound the diagnosis of dementia in this age group. A gradual age-related cognitive decline, particularly in executive function and mental speed, is evident even in non-demented oldest-old. Hearing and vision losses, which are also prevalent in the oldest-old and found in some cases to precede/predict cognitive decline, may mechanically interfere in neuropsychological evaluations. Difficulties in carrying out everyday activities, observed in the majority of the oldest-old, may be the result of motor or physical dysfunction and of neurodegenerative processes. The oldest-old appear to be a select population, who escapes major illnesses or delays their onset and duration toward the end of life. Dementia in the oldest-old may be manifested when a substantial amount of pathology is accumulated, or with a composition of a variety of pathologies. Investigating the clinical and pathological features of dementia in the oldest-old is of great importance in order to develop therapeutic strategies and to provide the most elderly of our population with good quality of life.


Thermodynamic propensities of amino acids in the native state ensemble: implications for fold recognition.
An amino acid sequence, in the context of the solvent environment, contains all of the thermodynamic information necessary to encode a three-dimensional protein structure. To investigate the relationship between an amino acid sequence and its corresponding protein fold, a database of thermodynamic stability information was assembled that spanned 2951 residues from 44 nonhomologous proteins. This information was obtained using the COREX algorithm, which computes an ensemble-based description of the native state of a protein. It was observed that amino acid types partitioned unequally into high, medium, and low thermodynamic stability environments. Furthermore, these distributions were reproducible and were significantly different than those expected from random partitioning. To assess the structural importance of the distributions, simple fold-recognition experiments were performed based on a 3D-1D scoring matrix containing only COREX residue stability information. This procedure was able to recover amino acid sequences corresponding to correct target structures more effectively than scoring matrices derived from randomized data. High-scoring sequences were often aligned correctly with their corresponding target profiles, suggesting that calculated thermodynamic stability profiles have the potential to encode sequence information. As a control, identical fold-recognition experiments were performed on the same database of proteins using DSSP secondary structure information in the scoring matrix, instead of COREX residue stability information. The comparable performance of both approaches suggested that COREX residue stability information and secondary structure information could be of equivalent utility in more sophisticated fold-recognition techniques. The results of this work are a consequence of the idea that amino acid sequences fold not into single, rigidly stable structures but rather into thermodynamic ensembles best represented by a time-averaged structure.


Profiling the Oxylipin and Endocannabinoid Metabolome by UPLC-ESI-MS/MS in Human Plasma to Monitor Postprandial Inflammation.
Bioactive lipids, including oxylipins, endocannabinoids, and related compounds may function as specific biochemical markers of certain aspects of inflammation. However, the postprandial responsiveness of these compounds is largely unknown; therefore, changes in the circulating oxylipin and endocannabinoid metabolome in response to a challenge meal were investigated at six occasions in a subject who freely modified her usual diet. The dietary change, and especially the challenge meal itself, represented a modification of precursor fatty acid status, with expectedly subtle effects on bioactive lipid levels. To detect even the slightest alteration, highly sensitive ultra-performance liquid chromatography (UPLC) coupled to electrospray ionization (ESI) tandem mass spectrometry (MS/MS) methods for bioactive lipid profiling was employed. A previously validated UPLC-ESI-MS/MS method for profiling the endocannabinoid metabolome was used, while validation of an UPLC-ESI-MS/MS method for oxylipin analysis was performed with acceptable outcomes for a majority of the parameters according to the US Food and Drug Administration guidelines for linearity (0.9938 < R2 < 0.9996), limit of detection (0.0005-2.1 pg on column), limit of quantification (0.0005-4.2 pg on column), inter- and intraday accuracy (85-115%) and precision (< 5%), recovery (40-109%) and stability (40-105%). Forty-seven of fifty-two bioactive lipids were detected in plasma samples at fasting and in the postprandial state (0.5, 1, and 3 hours after the meal). Multivariate analysis showed a significant shift of bioactive lipid profiles in the postprandial state due to inclusion of dairy products in the diet, which was in line with univariate analysis revealing seven compounds (NAGly, 9-HODE, 13-oxo-ODE, 9(10)-EpOME, 12(13)-EpOME, 20-HETE, and 11,12-DHET) that were significantly different between background diets in the postprandial state (but not at fasting). The only change in baseline levels at fasting was displayed by TXB2. Furthermore, postprandial responsiveness was detected for seven compounds (POEA, SEA, 9(10)-DiHOME, 12(13)-DiHOME, 13-oxo-ODE, 9-HODE, and 13-HODE). Hence, the data confirm that the UPLC-ESI-MS/MS method performance was sufficient to detect i) a shift, in the current case most notably in the postprandial bioactive lipid metabolome, caused by changes in diet and ii) responsiveness to a challenge meal for a subset of the oxylipin and endocannabinoid metabolome. To summarize, we have shown proof-of-concept of our UPLC-ESI-MS/MS bioactive lipid protocols for the purpose of monitoring subtle shifts, and thereby useful to address lipid-mediated postprandial inflammation.


Generating stochastic dispersed and periodic clustered textures using a composite hybrid screen.
In electrophotographic printing, a periodic clustered-dot halftone pattern is preferred for a smooth and stable result. In addition, the screen frequency should be high enough to minimize the visibility of the halftone textures and to ensure good detail rendition. However, at these frequencies, the halftone cell may contain too few pixels to provide a sufficient number of distinct gray levels. This will result in contouring and posterization. The traditional solution is to grow the clusters asynchronously within a repeating block of clusters known as a supercell. The growth of each individual cluster is governed by a microscreen. The order in which the clusters grow within the supercell is determined by a macroscreen. Typically, the macroscreen is a recursive pattern due to Bayer. In highlights and shadows, this ordering results in visible artifacts. Replacing the Bayer screen by a stochastic macroscreen eliminates these artifacts, but results in new artifacts. In this paper, we propose a new composite screen architecture that employs multiple microscreens and multiple macroscreens in the highlights and shadows. These screens are jointly designed by using the direct binary search (DBS) algorithm.


Milli-tesla NMR and spectrophotometry of liquids hyperpolarized by dissolution dynamic nuclear polarization.
Hyperpolarization methods offer a unique means of improving low signal strength obtained in low-field NMR. Here, simultaneous measurements of NMR at a field of 0.7mT and laser optical absorption from samples hyperpolarized by dissolution dynamic nuclear polarization (D-DNP) are reported. The NMR measurement field closely corresponds to a typical field encountered during sample injection in a D-DNP experiment. The optical spectroscopy allows determination of the concentration of the free radical required for DNP. Correlation of radical concentration to NMR measurement of spin polarization and spin-lattice relaxation time allows determination of relaxivity and can be used for optimization of the D-DNP process. Further, the observation of the nuclear Overhauser effect originating from hyperpolarized spins is demonstrated. Signals from (1)H and (19)F in a mixture of trifluoroethanol and water are detected in a single spectrum, while different atoms of the same type are distinguished by J-coupling patterns. The resulting signal changes of individual peaks are indicative of molecular contact, suggesting a new application area of hyperpolarized low-field NMR for the determination of intermolecular interactions.


Asthma and occupation in the 1958 birth cohort.
To examine the association of adult onset asthma with lifetime exposure to occupations and occupational exposures. We generated lifetime occupational histories for 9488 members of the British 1958 birth cohort up to age 42 years. Blind to asthma status, jobs were coded to the International Standard Classification of Occupations 1988 and an Asthma Specific Job Exposure Matrix (ASJEM) with an expert re-evaluation step. Associations of jobs and ASJEM exposures with adult onset asthma were assessed in logistic regression models adjusting for sex, smoking, social class at birth and childhood hay fever. Of the 7406 cohort members with no asthma or wheezy bronchitis in childhood, 639 (9%) reported asthma by age 42 years. Adult onset asthma was associated with 18 occupations, many previously identified as risks for asthma (eg, farmers: OR 4.26, 95% CI 2.06 to 8.80; hairdressers: OR 1.88, 95% CI 1.24 to 2.85; printing workers: OR 3.04, 95% CI 1.49 to 6.18). Four were cleaning occupations and a further three occupations were likely to use cleaning agents. Adult onset asthma was associated with five of the 18 high-risk specific ASJEM exposures (flour exposure: OR 2.12, 95% CI 1.17 to 3.85; enzyme exposure: OR 2.32, 95% CI 1.22 to 4.42; cleaning/disinfecting products: OR 1.67, 95% CI 1.26 to 2.22; metal and metal fumes: OR 1.45, 95% CI 1.02 to 2.07; textile production: OR 1.71, 95% CI 1.12 to 2.61). Approximately 16% (95% CI 3.8% to 27.1%) of adult onset asthma was associated with known asthmagenic occupational exposures. This study suggests that about 16% of adult onset asthma in British adults born in the late 1950s could be due to occupational exposures, mainly recognised high-risk exposures.


A novel treatment for ocular tumors using membrane FasL vesicles to activate innate immunity and terminate immune privilege.
Ocular immune privilege promotes tumor growth by hindering the development of innate and adaptive immunity. A prior study showed that ocular tumors expressing the membrane-only form of Fas ligand (FasL) terminate immune privilege, induce vigorous inflammation, undergo rejection, and induce systemic protective immunity. In these previous experiments the tumor cells used were genetically engineered to express membrane FasL. As an initial step toward developing an immunotherapy for intraocular tumors, the present study was conducted to examine whether injection of microvesicles expressing membrane FasL into ocular tumors (that are FasL negative) would have a similar effect. Microvesicles expressing either no FasL or membrane-only Fas ligand were coinjected with L5178Y-R lymphoma cells into the anterior chambers (AC) of DBA/2 mice. Tumor cells coinjected with control vesicles grew progressively in the AC, and all mice died of metastatic disease by day 15. By contrast, a single injection of membrane FasL vesicles induced a potent inflammatory response characterized by GR1+ neutrophils and F4/80+ macrophages and significantly improved survival from 0% in untreated mice to 58% in mFasL-treated mice. Among the surviving mice, the ocular tumor was eliminated in 55%, and the mice exhibited systemic protection from a second tumor challenge. In the remaining 45%, the ocular tumor was not eliminated, but the mice were protected from liver metastases. Bioactive membrane FasL microvesicles coinjected with tumor cells induce a potent inflammatory response that terminates immune privilege, eliminates ocular tumors, and prevents metastatic disease.


Round-the-clock power supply and a sustainable economy via synergistic integration of solar thermal power and hydrogen processes.
We introduce a paradigm-"hydricity"-that involves the coproduction of hydrogen and electricity from solar thermal energy and their judicious use to enable a sustainable economy. We identify and implement synergistic integrations while improving each of the two individual processes. When the proposed integrated process is operated in a standalone, solely power production mode, the resulting solar water power cycle can generate electricity with unprecedented efficiencies of 40-46%. Similarly, in standalone hydrogen mode, pressurized hydrogen is produced at efficiencies approaching ∼50%. In the coproduction mode, the coproduced hydrogen is stored for uninterrupted solar power production. When sunlight is unavailable, we envision that the stored hydrogen is used in a "turbine"-based hydrogen water power (H2WP) cycle with the calculated hydrogen-to-electricity efficiency of 65-70%, which is comparable to the fuel cell efficiencies. The H2WP cycle uses much of the same equipment as the solar water power cycle, reducing capital outlays. The overall sun-to-electricity efficiency of the hydricity process, averaged over a 24-h cycle, is shown to approach ∼35%, which is nearly the efficiency attained by using the best multijunction photovoltaic cells along with batteries. In comparison, our proposed process has the following advantages: (i) It stores energy thermochemically with a two- to threefold higher density, (ii) coproduced hydrogen has alternate uses in transportation/chemical/petrochemical industries, and (iii) unlike batteries, the stored energy does not discharge over time and the storage medium does not degrade with repeated uses.


Slow GABA transient and receptor desensitization shape synaptic responses evoked by hippocampal neurogliaform cells.
The kinetics of GABAergic synaptic currents can vary by an order of magnitude depending on the cell type. The neurogliaform cell (NGFC) has recently been identified as a key generator of slow GABA(A) receptor-mediated volume transmission in the isocortex. However, the mechanisms underlying slow GABA(A) receptor-mediated IPSCs and their use-dependent plasticity remain unknown. Here, we provide experimental and modeling data showing that hippocampal NGFCs generate an unusually prolonged (tens of milliseconds) but low-concentration (micromolar range) GABA transient, which is responsible for the slow response kinetics and which leads to a robust desensitization of postsynaptic GABA(A) receptors. This strongly contributes to the use-dependent synaptic depression elicited by various patterns of NGFC activity including the one detected during theta network oscillations in vivo. Synaptic depression mediated by NGFCs is likely to play an important modulatory role in the feedforward inhibition of CA1 pyramidal cells provided by the entorhinal cortex.


Sex Chromosome-wide Transcriptional Suppression and Compensatory Cis-Regulatory Evolution Mediate Gene Expression in the Drosophila Male Germline.
The evolution of heteromorphic sex chromosomes has repeatedly resulted in the evolution of sex chromosome-specific forms of regulation, including sex chromosome dosage compensation in the soma and meiotic sex chromosome inactivation in the germline. In the male germline of Drosophila melanogaster, a novel but poorly understood form of sex chromosome-specific transcriptional regulation occurs that is distinct from canonical sex chromosome dosage compensation or meiotic inactivation. Previous work shows that expression of reporter genes driven by testis-specific promoters is considerably lower-approximately 3-fold or more-for transgenes inserted into X chromosome versus autosome locations. Here we characterize this transcriptional suppression of X-linked genes in the male germline and its evolutionary consequences. Using transgenes and transpositions, we show that most endogenous X-linked genes, not just testis-specific ones, are transcriptionally suppressed several-fold specifically in the Drosophila male germline. In wild-type testes, this sex chromosome-wide transcriptional suppression is generally undetectable, being effectively compensated by the gene-by-gene evolutionary recruitment of strong promoters on the X chromosome. We identify and experimentally validate a promoter element sequence motif that is enriched upstream of the transcription start sites of hundreds of testis-expressed genes; evolutionarily conserved across species; associated with strong gene expression levels in testes; and overrepresented on the X chromosome. These findings show that the expression of X-linked genes in the Drosophila testes reflects a balance between chromosome-wide epigenetic transcriptional suppression and long-term compensatory adaptation by sex-linked genes. Our results have broad implications for the evolution of gene expression in the Drosophila male germline and for genome evolution.


Neuronal activity is required for the development of specific cortical interneuron subtypes.
Electrical activity has been shown to regulate development in a variety of species and in various structures, including the retina, spinal cord and cortex. Within the mammalian cortex specifically, the development of dendrites and commissural axons in pyramidal cells is activity-dependent. However, little is known about the developmental role of activity in the other major cortical population of neurons, the GABA-producing interneurons. These neurons are morphologically and functionally heterogeneous and efforts over the past decade have focused on determining the mechanisms that contribute to this diversity. It was recently discovered that 30% of all cortical interneurons arise from a relatively novel source within the ventral telencephalon, the caudal ganglionic eminence (CGE). Owing to their late birth date, these interneurons populate the cortex only after the majority of other interneurons and pyramidal cells are already in place and have started to functionally integrate. Here we demonstrate in mice that for CGE-derived reelin (Re)-positive and calretinin (Cr)-positive (but not vasoactive intestinal peptide (VIP)-positive) interneurons, activity is essential before postnatal day 3 for correct migration, and that after postnatal day 3, glutamate-mediated activity controls the development of their axons and dendrites. Furthermore, we show that the engulfment and cell motility 1 gene (Elmo1), a target of the transcription factor distal-less homeobox 1 (Dlx1), is selectively expressed in Re(+) and Cr(+) interneurons and is both necessary and sufficient for activity-dependent interneuron migration. Our findings reveal a selective requirement for activity in shaping the cortical integration of specific neuronal subtypes.


Evaluation of asymmetric dimethylarginine, nitric oxide levels and associated independent variables in obese and lean patients with polycystic ovarian syndrome.
To evaluate the asymmetric dimethylarginine (ADMA) and nitric oxide (NO) levels in obese and lean patients with polycystic ovarian syndrome (PCOS) and find out their relation with hormonal and metabolic parameters. Twenty-two obese, 18 lean patients with PCOS and 11 obese, 24 lean healthy control patients were enrolled prospectively. Plasma ADMA and NO levels and arginine/ADMA ratio were evaluated on 3rd day of menstrual cycle after at least 10 h overnight fasting. Plasma ADMA, NO levels and arginine/ADMA ratio were similar in the groups. ADMA level did not correlate with the hormonal and metabolic parameters in patients with PCOS. However, NO correlated inversely with fasting insulin (r = -0.353, p = 0.041) and homeostasis model of insulin resistance (HOMA-IR) (r = -0.379, p = 0.027). Arginine/ADMA ratio also correlated inversely with fasting insulin (r = -0.339, p = 0.050). In multinomial regression analysis the risk of low NO was associated independently with high fasting insulin (OR = 1.19, 95% CI 1.001-1.42, p = 0.049) and high HOMA-IR in patients with PCOS (OR = 2.26, 95% CI 1.03-4.98, p = 0.042). Insulin resistance may be the underlying mechanism of endothelial dysfunction through NO pathway in PCOS.


Identification and validation of a logistic regression model for predicting serious injuries associated with motor vehicle crashes.
A multivariate logistic regression model, based upon National Automotive Sampling System Crashworthiness Data System (NASS-CDS) data for calendar years 1999-2008, was developed to predict the probability that a crash-involved vehicle will contain one or more occupants with serious or incapacitating injuries. These vehicles were defined as containing at least one occupant coded with an Injury Severity Score (ISS) of greater than or equal to 15, in planar, non-rollover crash events involving Model Year 2000 and newer cars, light trucks, and vans. The target injury outcome measure was developed by the Centers for Disease Control and Prevention (CDC)-led National Expert Panel on Field Triage in their recent revision of the Field Triage Decision Scheme (American College of Surgeons, 2006). The parameters to be used for crash injury prediction were subsequently specified by the National Expert Panel. Model input parameters included: crash direction (front, left, right, and rear), change in velocity (delta-V), multiple vs. single impacts, belt use, presence of at least one older occupant (≥ 55 years old), presence of at least one female in the vehicle, and vehicle type (car, pickup truck, van, and sport utility). The model was developed using predictor variables that may be readily available, post-crash, from OnStar-like telematics systems. Model sensitivity and specificity were 40% and 98%, respectively, using a probability cutpoint of 0.20. The area under the receiver operator characteristic (ROC) curve for the final model was 0.84. Delta-V (mph), seat belt use and crash direction were the most important predictors of serious injury. Due to the complexity of factors associated with rollover-related injuries, a separate screening algorithm is needed to model injuries associated with this crash mode.


Recent advances in cell-mediated nanomaterial delivery systems for photothermal therapy.
Nanomaterial-based photothermal therapy using near-infrared (NIR) light is a promising strategy for cancer treatment. However, delivery of nano-agents to specific tumor sites poses a great challenge, especially in the tumor hypoxia regions, where the hypoxia-associated drug delivery barriers prevent the effective intratumoral distribution of nanomaterials. In this respect, tumor-tropic cells including macrophages, monocytes, neural stem cells (NSCs), and mesenchymal stem cells (MSCs) have been proposed as "Trojan Horse" delivery vehicles to transport various nano-agents to overcome the drug delivery barriers, increase the tumor targeting ability of nano-agents, and enhance the efficiency of photothermal therapy. In this review, we summarize recent research activities and emerging trends in this field, describe the advantages of combining cellular therapies and nanotechnology to achieve more effective cancer treatment, and discuss the clinical prospects of photothermal therapy.


Oxazaphosphorine cytostatics: past-present-future. Seventh Cain Memorial Award lecture.
The development of the oxazaphosphorine cytostatics cyclophosphamide, ifosfamide, and trofosfamide was based on the idea of applying the transport form/active form principle to the highly reactive nitrogen mustard group. A critical analysis and synopsis of the available results and knowledge will include examination of the extent to which the hypotheses on which this concept is based have been confirmed by experimental and clinical findings: 1. Chemical synthesis succeeded in converting the reactive nitrogen mustard into an inactive transport form (latentiation). 2. The requirement that the transport form be enzymatically activated to the active form in the target organ (the cancer cell) has been achieved by a sequence of metabolic reactions. 3. The aim of considerably increasing the therapeutic index of alkylating agents has been achieved by the oxazaphosphorine cytostatics. The greater cancerotoxic selectivity is closely correlated with the cytotoxic specificity of their activated primary metabolites. 4. The cancerotoxic selectivity of oxazaphosphorines was further increased when mesna was introduced as a regional uroprotector. Mesna eliminates the risk of therapy-limiting urotoxic side effects of oxazaphosphorines. With mesna protection, these cytostatics can be given in higher doses with increased safety, and their therapeutic efficacy can be enhanced. 5. Stabilization of the primary oxazaphosphorines, e.g., by attaching 2-mercaptoethanesulfonic acid (mafosfamide), opens up new possibilities in preclinical investigations and in therapy, e.g., for the clonogenic stem cell assay, for in vitro purging in autologous bone marrow transplantation, for regional perfusion of tumors, and, in small doses, for immunomodulation, where appropriate, in conjunction with "biological response modifiers."


[Inhaled budesonide for chronic obstructive pulmonary disease].
A significant, large minority of patients with chronic obstructive pulmonary disease (COPD) respond favorably to corticosteroid treatment; but the benefit may be outweighed by its side effects. Long-term administration of inhaled steroids is a safe means of treatment. We hypothesized that treatment with inhaled budesonide would improve clinical symptoms and pulmonary function in subjects with COPD, and that the response to an inhaled B2-agonist would individualize steroid responders. In 44 patients with stable COPD in a double- blind crossover trial, we compared a 6-week course of inhalations of 800 micrograms/d budesonide with a placebo, separated by a 4-week interval when no medication was taken. In 33 out of 42 responders to the B2-agonist who remained in the study, there was a significant improvement in FEV1 of greater than 20% following budesonide inhalation, as compared to placebo. There was also a significant difference between the 2 periods of treatment as to the mean number of B2-agonist inhalations. We conclude that about 1/4 of patients with stable COPD respond to bronchodilators, and treatment with inhaled steroids improves spirometry data and inhaled B2-agonist consumption in about 3/4.


Six-minute walk distance is not related to quality of life in patients with non-cystic fibrosis bronchiectasis.
To evaluate physical performance on the six-minute walk test (6MWT) in patients with non-cystic fibrosis bronchiectasis and to investigate its relationship with quality of life (QoL). To identify predictors of exercise performance, we also investigated whether six-minute walk distance (6MWD) is associated with clinical and spirometric findings. This was a cross-sectional study involving patients with non-cystic fibrosis bronchiectasis (age, > 18 years), with at least one respiratory symptom for > 2 years and an FEV1 < 70% of predicted. Patients underwent clinical evaluation, pulmonary function tests, the 6MWT, and QoL assessment with the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36). We included 70 patients (48 females). Mean age was 54.5 ± 17.7 years, and mean FEV1 was 44.9 ± 14.5% of predicted. The patients were divided into two groups: 6MWD-low (6MWD below the predicted lower limit; n = 23); and 6MWD-norm (normal 6MWD; n = 47). The following variables were significantly lower in the 6MWD-low group than in the 6MWD-norm group: age; age at diagnosis of bronchiectasis; proportion of former smokers; body mass index (BMI); FEV1% of predicted; and MEP% of predicted. There were no significant differences in the SF-36 scores between the groups. In the logistic regression model, lower age and lower BMI were significantly associated with lower 6MWD. In this sample, there was a high proportion of patients who presented a lower than expected 6MWD. Although 6MWD was not related to QoL, it was associated with age and BMI.


No effect of an UMTS mobile phone-like electromagnetic field of 1.97 GHz on human attention and reaction time.
Several studies in the past reported influences of electromagnetic emissions of GSM phones on reaction time in humans. However, there are currently only a few studies available dealing with possible effects of the electromagnetic fields emitted by UMTS mobile phones. In our study, 40 healthy volunteers (20 female, 20 male), aged 26.0 years (range 21-30 years) underwent four different computer tests measuring reaction time and attention under three different UMTS mobile phone-like exposure conditions (two exposure levels plus sham exposure). Exposure of the subjects was accomplished by small helical antennas operated close to the head and fed by a generic signal representing the emissions of a UMTS mobile phone under constant receiving conditions as well as under a condition of strongly varying transmit power. In the high exposure condition the resulting peak spatial average exposure of the test subjects in the cortex of the left temporal lobe of the brain was 0.63 W/kg (min. 0.25 W/kg, max. 1.49 W/kg) in terms of 1 g averaged SAR and 0.37 W/kg (min. 0.16 W/kg, max. 0.84 W/kg) in terms of 10 g averaged SAR, respectively. Low exposure condition was one-tenth of high exposure and sham was at least 50 dB below low exposure. Statistical analysis of the obtained test parameters showed that exposure to the generic UMTS signal had no statistically significant immediate effect on attention or reaction. Therefore, this study does not provide any evidence that exposure of UMTS mobiles interferes with attention under short-term exposure conditions.


Performance in multiple domains of social cognition in parents of patients with schizophrenia.
Social cognition refers to a set of cognitive abilities that allow us to perceive and interpret social stimuli. Social cognition is affected in schizophrenia and impairments have also been documented in unaffected relatives, suggesting that social cognition may be related to a genetic vulnerability to the disease. This study aims to investigate potential impairments in four domains of social cognition (mentalizing, emotion recognition, social knowledge and empathy) in the same group of relatives in order to gather a more complete picture of social cognition difficulties in this population. The Batterie Intégrée de Cognition Sociale (BICS) (mentalizing, emotion recognition, and social knowledge) and the Interpersonal Reactivity Index (IRI) (empathy) were administered to 31 parents of patients with a psychotic disorder and 38 healthy controls. Parents of patients performed significantly worse than controls on the mentalizing test but significantly better on the social knowledge test. No significant between-group differences were observed for emotion recognition and empathy. This study is the first to evaluate four social cognition domains in this population. The results precise which social cognition processes may be impaired or preserved in unaffected relatives of patients and lead us to propose an hypothesis about a mechanism that could underlie the mentalizing difficulties observed in this population.


Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive.
Taurine is a biologically and physiologically valuable food additive. However, commercial taurine production mainly relies on environmentally harmful chemical synthesis. Herein, for the first time in bacteria, we attempted to produce taurine in metabolically engineered Corynebacterium glutamicum. The taurine-producing strain was developed by introducing cs, cdo1, and csad genes. Interestingly, while the control strain could not produce taurine, the engineered strains successfully produced taurine via the newly introduced metabolic pathway. Furthermore, we investigated the effect of a deletion strain of the transcriptional repressor McbR gene on taurine production. As a result, sulfur accumulation and l-cysteine biosynthesis were reinforced by the McbR deletion strain, which further increased the taurine production by 2.3-fold. Taurine production of the final engineered strain Tau11 was higher than in other previously reported strains. This study demonstrated a potential approach for eco-friendly biosynthesis as an alternative to the chemical synthesis of a food additive.


A description of the methods of the aspirin supplementation for pregnancy indicated risk reduction in nulliparas (ASPIRIN) study.
Preterm birth (PTB) remains the leading cause of neonatal mortality and long term disability throughout the world. Though complex in its origins, a growing body of evidence suggests that first trimester administration of low dose aspirin (LDA) may substantially reduce the rate of PTB. Hypothesis: LDA initiated in the first trimester reduces the risk of preterm birth. Study Design Type: Prospective randomized, placebo-controlled, double-blinded multi-national clinical trial conducted in seven low and middle income countries. Trial will be individually randomized with one-to-one ratio (intervention/control) Population: Nulliparous women between the ages of 14 and 40, with a singleton pregnancy between 6 0/7 weeks and 13 6/7 weeks gestational age (GA) confirmed by ultrasound prior to enrollment, no more than two previous first trimester pregnancy losses, and no contraindications to aspirin. Daily administration of low dose (81 mg) aspirin, initiated between 6 0/7 weeks and 13 6/7 weeks GA and continued to 36 0/7 weeks GA, compared to an identical appearing placebo. Compliance and outcomes will be assessed biweekly. Primary outcome: Incidence of PTB (birth prior to 37 0/7 weeks GA). Secondary outcomes Incidence of preeclampsia/eclampsia, small for gestational age and perinatal mortality. This study is unique as it will examine the impact of LDA early in pregnancy in low-middle income countries with preterm birth as a primary outcome. The importance of developing low-cost, high impact interventions in low-middle income countries is magnified as they are often unable to bear the financial costs of treating illness. ClinicalTrials.gov identifier: NCT02409680 Date: March 30, 2015.


Effect of aqueous humor on apoptosis of inflammatory cell types.
To determine whether aqueous humor promotes cell death in cells involved in inflammatory responses. Multiple immune cell types, most characteristically involved in inflammatory responses, were incubated for 24, 48, and 72 hours in the presence or absence of 50% aqueous humor. Promotion of cell death was assayed by staining for an early indicator of apoptosis. The percentage of cells undergoing apoptosis was measured by flow cytometry. To identify partially the apoptosis inducing factor, aqueous humor was pretreated with proteinase K to degrade protein. In other experiments, aqueous humor was fractionated by centrifugation on filters capable of separating molecules above and below 10 kDa or 30 kDa kilodaltons in size. Rabbit aqueous humor promoted apoptosis in a wide variety of immune cells, including lymphokine-activated natural killer cells, resting T cells, an activated T-cell line, RAW 264.7 and J774A0.1 monocyte-macrophage cell lines, and neutrophils. As previously shown, aqueous humor did not promote apoptosis of murine corneal endothelial cells. Apoptosis was also not induced in human corneal endothelium, mouse corneal epithelium, or iris/ciliary body cell lines. Instead, aqueous humor partially protected these ocular tissues from starvation-induced cell death. Pretreatment with proteinase K inhibited the apoptosis-inducing activity. Moreover, the apoptosis-inducing activity segregated with the aqueous humor fraction containing molecules less than than 10 kDa in size. These data show that aqueous humor contains a factor or factors that promote death of cells that participate in inflammatory processes. By contrast, ocular tissues, such as the corneal endothelium and iris/ciliary body, are impervious to aqueous humor-induced cell death. The aqueous humor- borne factor(s) may contribute to the immune privilege of the anterior chamber by purging potential inflammatory cells.


Evolutionary history and attenuation of myxoma virus on two continents.
The attenuation of myxoma virus (MYXV) following its introduction as a biological control into the European rabbit populations of Australia and Europe is the canonical study of the evolution of virulence. However, the evolutionary genetics of this profound change in host-pathogen relationship is unknown. We describe the genome-scale evolution of MYXV covering a range of virulence grades sampled over 49 years from the parallel Australian and European epidemics, including the high-virulence progenitor strains released in the early 1950s. MYXV evolved rapidly over the sampling period, exhibiting one of the highest nucleotide substitution rates ever reported for a double-stranded DNA virus, and indicative of a relatively high mutation rate and/or a continually changing selective environment. Our comparative sequence data reveal that changes in virulence involved multiple genes, likely losses of gene function due to insertion-deletion events, and no mutations common to specific virulence grades. Hence, despite the similarity in selection pressures there are multiple genetic routes to attain either highly virulent or attenuated phenotypes in MYXV, resulting in convergence for phenotype but not genotype.


Controlling band alignments by artificial interface dipoles at perovskite heterointerfaces.
The concept 'the interface is the device' is embodied in a wide variety of interfacial electronic phenomena and associated applications in oxide materials, ranging from catalysts and clean energy systems to emerging multifunctional devices. Many device properties are defined by the band alignment, which is often influenced by interface dipoles. On the other hand, the ability to purposefully create and control interface dipoles is a relatively unexplored degree of freedom for perovskite oxides, which should be particularly effective for such ionic materials. Here we demonstrate tuning the band alignment in perovskite metal-semiconductor heterojunctions over a broad range of 1.7 eV. This is achieved by the insertion of positive or negative charges at the interface, and the resultant dipole formed by the induced screening charge. This approach can be broadly used in applications where decoupling the band alignment from the constituent work functions and electron affinities can enhance device functionality.


Anti-Ro/SSA antibodies. Association with a particulate (large speckledlike thread) immunofluorescent nuclear staining pattern.
Twenty-one patients with clinical and histopathologic evidence of subacute cutaneous lupus erythematosus and one patient with Sjögren's syndrome and vasculitis had anti-Ro/SSA (Sjögren's syndrome A) antibodies as demonstrated by double immunodiffusion assay using a saline extract of human spleen as the source of antigen. These serum samples were negative when tested for other nuclear antigens, including native DNA, Sm, and RNP. When tested for antinuclear antibodies (ANAs) by immunofluorescence, only ten of 22 serum samples were ANA positive on mouse liver substrate, while 18 of 22 had a positive ANA when HEp-2 tumor cells were utilized. With imprints of human spleen as test substrate, all 22 serum samples yielded a positive ANA result and a particulate (large speckledlike thread) staining pattern. Absorption of two of these serum samples with human spleen extract containing Ro/SSA antigen inhibited both the particulate staining pattern using human spleen imprints and the anti-Ro/SSA precipitin line by double immunodiffusion. These studies suggest that anti-Ro/SSA antibodies and antibodies producing the particulate nuclear staining pattern on human spleen imprints are either one and the same or closely paralleling antibody systems.


The role of CD4(+) T cells in BKV-specific T cell immunity.
Reactivation of polyomavirus BK (BKV) infection represents a severe complication in kidney transplant (KTX) patients. We previously reported an association between a declining BK viral load and the reconstitution of CD4(+) T cell BKV-specific immunity in patients following kidney transplantation. However, the specific contribution of CD4(+) T cells in the regulation of BKV-replication is unknown. Nevertheless, in vitro enrichment of BKV-specific T cells and subsequent adoptive T cell transfer may improve the restoration of immune competence in KTX patients with BKV infection. To date, strategies to capture human BKV-specific T cells with the ensuing expansion to clinically useful numbers are lacking. Here, we demonstrated a comprehensive flow cytometric analysis of the BKV-specific T cell response that permits access to the majority of T cells specific for immunodominant BKV antigens. A full-spectrum evaluation of the BKV-specific T cell response was performed by stimulating peripheral blood mononuclear cells (PBMC) with a mixture of BKV immunodominant peptide pools at varying concentrations and measuring activation marker expression and cytokine secretion. We also examined the effects of co-stimulation and PBMC resting time prior to activation. We defined the narrow range of stimulation conditions that permit the capture and expansion of functional BKV-specific T cell lines. The generated BKV-specific T cell lines showed the highest specificity and functionality when the T cells were captured according to IFNγ-secretion. This study highlights the multifunctional and cytolytic BKV-specific CD4(+) T cells as a dominant population within the generated T cell product. This method offers a novel approach for the generation of BKV-specific T cell lines for adoptive immunotherapy and underscores the critical role of CD4(+) T cells in the clearance of BKV.


Effect of physical activity on nonmelanoma skin cancer risk in kidney, liver, and pancreatic transplant patients.
Physical activity has been associated with a lower risk of cancer, including melanoma, but the effect has not yet been assessed for nonmelanoma skin cancer (NMSC). To determine whether the risk of skin cancer differs according to physical activity level in patients at high risk for NMSC. We conducted a retrospective cohort study in kidney, liver, and pancreatic transplant patients via telephone interview. Physical activity scores were calculated for each patient using a previously validated questionnaire. Sun exposure history and skin type were also recorded. The outcome of interest was a biopsy-proven diagnosis of at least one NMSC. Of 142 subjects, 45 (32%) developed NMSC. There was no significant effect of physical activity on the development of NMSC. Physical activity level may not be a major predictor of skin cancer risk in organ transplant patients. Controlled trials and population-based studies are needed to determine whether exercise can decrease the risk of NMSC in the general population. The authors have indicated no significant interest with commercial supporters.


A neonatal mouse model of intestinal perforation: investigating the harmful synergism between glucocorticoids and indomethacin.
Early postnatal steroids and indomethacin in combination have been shown to increase the risk of spontaneous intestinal perforation (SIP) in infants with extremely low birth weight (ELBW), but the mechanism behind this synergistic effect is unknown. Based on literature in a variety of models suggesting that glucocorticoids and nonsteroidal antiinflammatory agents diminish complementary isoforms of nitric oxide synthase (NOS), we hypothesized that perturbations in NO metabolism contribute to SIP. Our results using newborn wild-type (WT) and endothelial NOS-knockout (eNOS KO) mice treated with dexamethasone and/or indomethacin indicate that indomethacin treatment diminishes ileal eNOS abundance; dexamethasone treatment diminishes ileal inducible NOS and neuronal NOS (nNOS); 100% of dexamethasone-treated eNOS KO mice die after 3 days; eNOS KO mice treated for 2 days with dexamethasone develop acute pyloric stenosis in association with reduced expression of pyloric nNOS; and isolated ileum from eNOS KO mice treated for 2 days with dexamethasone exhibit a significant decrease in spontaneous peristalsis, decreased circumference, and decreased capacitance for forced volume before ileal perforation compared with ileum from untreated controls. Our findings suggest that eNOS and nNOS display functional overlap in the newborn mouse gastrointestinal tract and that simultaneous reduction in the activity of both NOS isoforms may be a risk factor for neonatal ileal perforation. If this holds true in human infants, then it provides a plausible etiologic explanation for the strong temporal association between SIP and the simultaneous treatment of ELBW infants with glucocorticoids and indomethacin.


Fibrous radial structure of speckle patterns in polychromatic light.
A fibrous radial structure of speckle patterns produced at the far-field diffraction plane by various diffuse objects under illumination of polychromatic light is studied theoretically and experimentally by means of the spatial correlation function of the speckle intensity. It is shown that the fibrous radial structure of speckle patterns strongly depends on both the surface roughness of diffuse objects under illumination and the radial distance from the center of patterns at the far-field diffraction plane.


Symptomatic unruptured cerebral aneurysms: features and surgical outcome.
Development of less invasive imaging studies, such as magnetic resonance angiography, has increased the chances that unruptured cerebral aneurysms are found. The rupture risk of "symptomatic" aneurysms is higher than for "asymptomatic" aneurysms; so "symptomatic" aneurysms are more often surgically treated. Many reviews examine "asymptomatic" unruptured cerebral aneurysms, but few evaluate "symptomatic" aneurysms. The author has treated many patients with symptomatic unruptured cerebral aneurysms and found that improved cranial nerve signs can be expected if the surgical treatment is performed before the symptoms become irreversible; the critical period is approximately 3 months. It is important to suppress the pulsation of the aneurysms compressing the cranial nerves; both a clipping procedure and endovascular coiling are effective. Cranial nerve signs are more commonly the symptoms of unruptured cerebral aneurysms, but large to giant aneurysms can also be the causes of hemiparesis, hydrocephalus, epilepsy, or even cerebral infarction. This review summarizes the features and surgical outcome of symptomatic unruptured cerebral aneurysms.


Pathophysiology of prediabetes and treatment implications for the prevention of type 2 diabetes mellitus.
Type 2 diabetes and other non-communicable diseases (NCD) are a growing public health challenge globally. An estimated 285 million people, corresponding to 6.4 % of the world's adult population has diabetes. This is expected to reach 552 million by 2030, 7.8 % of the adult population, with the African region expected to experience the greatest increase. A much larger segment of the world's population, approximating 79 million individuals in the US alone, has prediabetes. Multiple factors including genetic predisposition, insulin resistance, increased insulin secretory demand, glucotoxicity, lipotoxicity, impaired incretin release/action, amylin accumulation, and decreased β-cell mass play a causative role in the progressive β-cell dysfunction characteristic of prediabetes. Interventions preventing progression to type 2 diabetes should therefore delay or prevent β-cell failure. This article will first review the principal pathophysiological mechanisms underlying prediabetes and subsequently address treatment considerations based on these in the prevention of type 2 diabetes. In view of long-standing safety data with demonstrated efficacy and cost-effectiveness in the prevention of type 2 diabetes in high-risk individuals, metformin should be considered as initial therapy for those unable to comply with or lifestyle modification or where the latter has been ineffective in decreasing progression to type 2 diabetes.


Mate Selection in Self-Compatible Wild Tobacco Results from Coordinated Variation in Homologous Self-Incompatibility Genes.
In flowering plants, intraspecific mate preference is frequently related to mating systems: the rejection of self pollen in self-incompatible (SI) plants that prevents inbreeding is one of the best described examples. However, in other mating systems, more nuanced patterns of pollen rejection occur. In the self-compatible (SC) Nicotiana attenuata, in which SI is not found and all crosses are compatible, certain pollen genotypes are consistently selected in mixed pollinations. However, the molecular mechanisms of this polyandrous mate selection remain unknown. Style-expressed NaS-like-RNases and pollen-expressed NaSLF-like genes, homologous to SI factors in Solanaceae, were identified and examined for a role in N. attenuata's mate selection. A comparison of two NaS-like-RNases and six NaSLF-like genes among 26 natural accessions revealed specific combinations of co-expression and direct protein-protein interactions. To evaluate their role in mate selection, we silenced the expression of specific NaS-like-RNases and NaSLF-like proteins and conducted diagnostic binary mixed pollinations and mixed pollinations with 14 different non-self pollen donors. Styles expressing particular combinations of NaS-like-RNases selected mates from plants with corresponding NaS-like-RNase expression patterns, while styles lacking NaS-like-RNase expression were non-selective in their fertilizations, which reflected the genotype ratios of pollen mixtures deposited on the stigmas. DNA methylation could account for some of the observed variation in stylar NaS-like-RNase patterns. We conclude that the S-RNase-SLF recognition mechanism plays a central role in polyandrous mate selection in this self-compatible species. These results suggest that after the SI-SC transition, natural variation of SI homologous genes was repurposed to mediate intraspecific mate selection.


Genome-Wide Analysis of Left Ventricular Maximum Wall Thickness in the UK Biobank Cohort Reveals a Shared Genetic Background With Hypertrophic Cardiomyopathy.
Left ventricular maximum wall thickness (LVMWT) is an important biomarker of left ventricular hypertrophy and provides diagnostic and prognostic information in hypertrophic cardiomyopathy (HCM). Limited information is available on the genetic determinants of LVMWT. We performed a genome-wide association study of LVMWT measured from the cardiovascular magnetic resonance examinations of 42 176 European individuals. We evaluated the genetic relationship between LVMWT and HCM by performing pairwise analysis using the data from the Hypertrophic Cardiomyopathy Registry in which the controls were randomly selected from UK Biobank individuals not included in the cardiovascular magnetic resonance sub-study. Twenty-one genetic loci were discovered at P<5×10-8. Several novel candidate genes were identified including PROX1, PXN, and PTK2, with known functional roles in myocardial growth and sarcomere organization. The LVMWT genetic risk score is predictive of HCM in the Hypertrophic Cardiomyopathy Registry (odds ratio per SD: 1.18 [95% CI, 1.13-1.23]) with pairwise analyses demonstrating a moderate genetic correlation (rg=0.53) and substantial loci overlap (19/21). Our findings provide novel insights into the genetic underpinning of LVMWT and highlight its shared genetic background with HCM, supporting future endeavours to elucidate the genetic etiology of HCM.


Caffeic acid phenethyl ester alleviates asthma by regulating the airway microenvironment via the ROS-responsive MAPK/Akt pathway.
In the pathophysiology of asthma, structural cell dysfunction and concomitant microenvironment changes in airways are crucial to pathological progression, which involves oxidative stress. Caffeic acid phenethyl ester (CAPE) is an active anti-oxidative component obtained from propolis, and has been shown to have beneficial effects on several respiratory disorders, such as chronic obstructive pulmonary disease and lung cancer. However, the impact of CAPE on asthma is not well understood. Therefore, this study investigated the advantages of using CAPE to treat asthma and demonstrated the roles of CAPE in the regulation of airway microenvironments. In ovalbumin (OVA)-sensitized mice, CAPE treatments notably reduced airway hyperresponsiveness, attenuated extensive inflammatory cell infiltration and inhibited goblet cell hyperplasia and collagen deposition and fibrosis. In addition, CAPE improved the airway microenvironment in a dose-dependent manner by inhibiting OVA-induced increases in immunoglobulin E, tumor necrosis factor alpha (TNF-α), transforming growth factor-β1 (TGF-β1), interleukin (IL)-4 and IL-13 and suppressing matrix metalloproteinase-9 and alpha-smooth muscle actin expression as well as malondialdehyde production. To determine the underlying mechanisms responsible for these effects, we used TNF-α-stimulated BECs and TGF-β1-challenged human ASMCs to explore the impacts of CAPE on pro-inflammatory proteins and ASMC proliferation. The results indicated that CAPE significantly limited the secretion of eotaxin-1, monocyte chemoattractant protein-1, IL-8 and intercellular adhesion molecule-1 and dramatically inhibited the proliferation of ASMCs. These effects were shown to be associated with decreased reactive oxidant species (ROS) levels. The phosphorylation of Akt and Mitogen-Activated Protein Kinase (MAPK) caused by increased ROS was significantly decreased by CAPE, which implied a contribution of ROS-MAPK/Akt signaling to the attenuation of asthma. Our findings indicated for the first time that CAPE alleviates airway inflammation and remodeling in chronic asthma by balancing the airway microenvironment, which highlights a novel profile of CAPE as a potent agent for asthma management.


Impairment-Based 3-D Robotic Intervention Improves Upper Extremity Work Area in Chronic Stroke: Targeting Abnormal Joint Torque Coupling With Progressive Shoulder Abduction Loading.
The implementation of a robotic system (ACT(3D)) that allowed for a quantitative measurement of abnormal joint torque coupling in chronic stroke survivors and, most importantly, a quantitative means of initiating and progressing an impairment-based intervention, is described. Individuals with chronic moderate to severe stroke (n = 8) participated in this single-group pretest-posttest design study. Subjects were trained over eight weeks by progressively increasing the level of shoulder abduction loading experienced by the participant during reaching repetitions as performance improved. Reaching work area was evaluated pre- and postintervention for ten different shoulder abduction loading levels along with isometric single-joint strength and a qualitative clinical assessment of impairment. There was a significant effect of session (pre versus post) with an increase in reaching work area, despite no change in single-joint strength. This data suggests that specifically targeting the abnormal joint torque coupling impairment through progressive shoulder abduction loading is an effective strategy for improving reaching work area following hemiparetic stroke. Application of robotics, namely, the ACT(3D), allowed for quantitative control of the exercise parameters needed to directly target the synergistic coupling impairment. The targeted reduction of abnormal joint torque coupling is likely the key factor explaining the improvements in reaching range of motion achieved with this intervention.


Coexpression and stimulation of parathyroid hormone receptor positively regulates slowly activating IsK channels expressed in Xenopus oocytes.
Expression of the IsK protein in Xenopus oocytes induced the characteristically slow, voltage-dependent outward currents. Superfusion with the parathyroid hormone (PTH) peptide 1-34 had no effect on IsK when expressed alone, but increased IsK when IsK was coexpressed with the PTH-receptor. PTH receptor stimulation caused a shift of IsK conductance-voltage relationship to more negative potentials, and a decrease of both the rate of IsK activation and deactivation. IsK regulation by PTH was independent of extracellular Ca2+, and was also present IsK protein mutants lacking the protein kinase C consensus site. However, regulation of IsK by PTH was mimicked by activators of protein kinase A (PKA) and greatly reduced in the presence of the kinase inhibitors staurosporine and H89. These results suggest that PTH regulates IsK by a mechanism involving phosphorylation independent of protein kinase C (PKC). Such regulation may play a role in proximal tubule cells of the kidney, where both PTH receptor and the IsK protein are expressed.


The relationship between nerve conduction velocity and fiber morphology during peripheral nerve regeneration.
We analyzed the relationship between motor nerve conduction velocity (MCV) and morphological changes in regenerating nerve fibers at different times after sciatic nerve transection to identify reliable indices of functional recovery. Thirty rats were divided into five equal groups, one control group and four groups subjected to sciatic nerve transection and immediate suturing, followed by regeneration for 50, 100, 150, and 200 days, respectively. MCV was measured in each group, followed by morphometric analyses of fibers of the common peroneal nerve. MCV increased progressively with time after nerve transection, although it remained lower than the control velocity. Mean fiber diameter (axon plus myelin sheath) also increased with time after nerve transection. Recovery of mean fiber diameter was well correlated with MCV, even though regenerating nerves likely contained many small nonconducting fibers. In contrast, the change in the mean diameter of regenerating axons and relative myelin thickness (g-ratio) did not provide an accurate measure of recovery as they were not increasing in a time-dependent manner. Furthermore, internodal length changed only slightly with increasing fiber diameter in regenerating nerves; therefore, the regression relation between fiber diameter and internodal length was not a sensitive index of recovery. MCV and mean fiber diameter were the most sensitive indices of functional recovery during sciatic nerve regeneration.


Thioridazine and sudden unexplained death in psychiatric in-patients.
Sudden death has been linked to antipsychotic therapy, but the relative risk associated with specific drugs is unknown. To assess the risk of sudden unexplained death associated with antipsychotic drug therapy and its relation to drug dose and individual agents. A case-control study of psychiatric in-patients dying suddenly in five hospitals in the north-east of England and surviving controls matched for age, gender and mental disorder. Logistic regression analysis was used to identify significant risk factors, and odds ratios were calculated. Sixty-nine case-control clusters were identified. Probable sudden unexplained death was significantly associated with hypertension, ischaemic heart disease and current treatment with thioridazine (adjusted odds ratio=5.3, 95% CI 1.7-16.2, P=0.004). There was no significant association with other individual antipsychotic drugs. Thioridazine alone was associated with sudden unexplained death, the likely mechanism being drug-induced arrhythmia.


Manipulating patterns of dynamic deformation elicits the impression of cloth with varying stiffness.
Cloth is a common material, and humans can visually estimate its mechanical properties by observing how it deforms under external forces. Here, we ask whether and how dynamic deformation can affect the perception of mechanical properties of cloth. In Experiment 1, we find that both intrinsic mechanical properties and optical properties affect stiffness perception when the stimuli are presented as images. By contrast, in videos, humans can partially discount the effect of optical appearances and exhibit higher sensitivity to stiffness. We further identified an idiosyncratic deformation pattern (i.e., movement uniformity) to differentiate stiffness, which can be reliably measured by six optical flow features. In Experiment 2, we isolate the deformation by creating dynamic dot stimuli from the 3-D mesh of the cloth. We directly alter the movement pattern by manipulating the uniformity of the displacement vectors on the dot stimuli and show that changing the pattern of dynamic deformation alone can alter the perceived stiffness of cloth in a variety of scene setups. Furthermore, by analyzing optical flow fields extracted from the manipulated dynamic dot stimuli, we confirmed the same six optical flow features can be diagnostic of the degree of stiffness of moving cloth across different scenes. Overall, our study demonstrates that manipulating patterns of dynamic deformation alone can elicit the impression of cloth with varying stiffness, suggesting that the human visual system might rely on the idiosyncratic pattern of dynamic deformation for estimating stiffness.


Image-Based Artefact Removal in Laser Scanning Microscopy.
Recent developments in laser scanning microscopy have greatly extended its applicability in cancer imaging beyond the visualization of complex biology, and opened up the possibility of quantitative analysis of inherently dynamic biological processes. However, the physics of image acquisition intrinsically means that image quality is subject to a tradeoff between a number of imaging parameters, including resolution, signal-to-noise ratio, and acquisition speed. We address the problem of geometric distortion, in particular, jaggedness artefacts that are caused by the variable motion of the microscope laser, by using a combination of image processing techniques. Image restoration methods have already shown great potential for post-acquisition image analysis. The performance of our proposed image restoration technique was first quantitatively evaluated using phantom data with different textures, and then qualitatively assessed using in vivo biological imaging data. In both cases, the presented method, comprising a combination of image registration and filtering, is demonstrated to have substantial improvement over state-of-the-art microscopy acquisition methods.


Efficacy of commonly used disinfectants for inactivation of human noroviruses and their surrogates.
Human noroviruses (HuNoVs) are the most common cause of acute viral gastroenteritis worldwide and are a leading cause of foodborne disease. Their environmental persistence and purported resistance to disinfection undoubtedly contribute to their success as foodborne disease agents. The purpose of this study was to compare the efficacy of three commonly used disinfectant active ingredients against representative HuNoV strains and cultivable surrogates. Ethanol (50, 70, and 90%), sodium hypochlorite (5, 75, 250, 500, and 1,000 ppm), and a quaternary ammonium compound blend (at 0.1×, 1.0×, and 10× concentrations) were evaluated against two norovirus (NoV) genogroup II strains (GII.2 and GII.4) and two surrogates (feline calicivirus [FCV] and murine norovirus [MNV-1]). Virucidal suspension assays (30-s exposure) were conducted in accordance with ASTM International standard E-1052. Virus inactivation was quantified using reverse transcription quantitative PCR targeting the ORFI-ORFII junction (HuNoV), the RNA polymerase region (MNV-1), or the ORFI region (FCV); infectivity assays were also performed for MNV-1 and FCV. The two HuNoV strains and FCV were relatively resistant to ethanol (<0.5 log inactivation) irrespective of concentration, but MNV-1 was much more susceptible (log inactivation, ∼2.0 log at higher ethanol concentrations). Both HuNoV strains were more resistant to hypochlorite than were either of the animal surrogates, with the human strains requiring ≥500 ppm of hypochlorite to achieve statistically significant reduction (≥3.0 log) in virus concentration. All four viruses were resistant to inactivation (<0.5-log reduction) using the quaternary ammonium compound formulation at all concentrations tested. This study is novel in that it clearly demonstrates the relative ineffectiveness of common active disinfectant ingredients against HuNoV and highlights the fact that the cultivable surrogates do not always mimic HuNoV strains.


Pleiotropic phenotypes of the sticky peel mutant provide new insight into the role of CUTIN DEFICIENT2 in epidermal cell function in tomato.
Plant epidermal cells have evolved specialist functions associated with adaptation to stress. These include the synthesis and deposition of specialized metabolites such as waxes and cutin together with flavonoids and anthocyanins, which have important roles in providing a barrier to water loss and protection against UV radiation, respectively. Characterization of the sticky peel (pe) mutant of tomato (Solanum lycopersicum) revealed several phenotypes indicative of a defect in epidermal cell function, including reduced anthocyanin accumulation, a lower density of glandular trichomes, and an associated reduction in trichome-derived terpenes. In addition, pe mutant fruit are glossy and peels have increased elasticity due to a severe reduction in cutin biosynthesis and altered wax deposition. Leaves of the pe mutant are also cutin deficient and the epicuticular waxes contain a lower proportion of long-chain alkanes. Direct measurements of transpiration, together with chlorophyll-leaching assays, indicate increased cuticular permeability of pe leaves. Genetic mapping revealed that the pe locus represents a new allele of CUTIN DEFICIENT2 (CD2), a member of the class IV homeodomain-leucine zipper gene family, previously only associated with cutin deficiency in tomato fruit. CD2 is preferentially expressed in epidermal cells of tomato stems and is a homolog of Arabidopsis (Arabidopsis thaliana) ANTHOCYANINLESS2 (ANL2). Analysis of cuticle composition in leaves of anl2 revealed that cutin accumulates to approximately 60% of the levels observed in wild-type Arabidopsis. Together, these data provide new insight into the role of CD2 and ANL2 in regulating diverse metabolic pathways and in particular, those associated with epidermal cells.


Proteomics analysis of human endothelial cell line EA.hy926 after exposure to GSM 900 radiation.
The human endothelial cell line EA.hy926 was exposed to mobile phone radiation and the effect on protein expression was examined using two-dimensional electrophoresis (2-DE). Up to 38 various proteins have statistically significantly altered their expression levels following the irradiation. Four proteins were identified with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). Two of the affected proteins were determined to be isoforms of cytoskeletal vimentin. This finding supports our earlier presented working hypothesis which indicated that the mobile phone radiation might affect the cytoskeleton and might have an effect on the physiological functions that are regulated by the cytoskeleton.


Self-assembled organometallic [12]metallacrown-3 complexes.
The reaction of [(arene)RuCl2]2 (arene = C6H6, cymene, C6H3Et3, or C6Me6) or [Cp*RhCl2]2 with 3-hydroxy-2-pyridone in the presence of Cs2CO3 gives trinuclear metallamacrocyclic complexes. The self-assembly process was shown to be completely diastereoselective, and a racemic mixture of complexes with M(R)M(R)M(R) or MsMsMs (M=Ru, Rh) configuration was obtained. Plausible mononuclear intermediates of the formula [(arene)RuCl(C5H4NO2)] (arene = cymene, C6Me6) have been isolated and characterized. A structurally related trimer was synthesized by using [(cymene)RuCl2]2 and 3-acetamido-2-pyridone instead of 3-hydroxy-2-pyridone. The macrocycles were shown to be highly potent ionophores for Na+ and/or Li+ with negligible affinities for the larger cation K+. The selectivities of the receptors depend on the pi-ligand present: whereas the (C6H6)Ru- and (cymene)Ru complexes bind both Li+ and Na+, the (C6Me6)Ru-, (C6H3Et3)Ru-, and Cp*Rh complexes bind exclusively Li+. For all receptors, the presence of alkali metal ions can be detected electrochemically: the peak potential is shifted by > 300 mV toward anionic potential upon binding. This behavior was utilized to detect Li+ and Na+ colorimetrically. Single crystal X-ray analyses have been carried out on eight complexes, four of which are bound to an alkali metal halide ion pair. Structural parameters, which affect the affinity and selectivity are discussed. A computational study on [[MX][12]crown-3] complexes (M =Li, Na; X=Cl, Br, I) was performed in order to compare relevant bond lengths and angles of the energy-minimized structures with those of the organometallic receptors.


Hypothetical midlife interventions in women and risk of type 2 diabetes.
Randomized trials have examined short-term effects of lifestyle interventions for diabetes prevention only among high-risk individuals. Prospective studies have examined the associations between lifestyle factors and diabetes in healthy populations but have not characterized the intervention. We estimated the long-term effects of hypothetical lifestyle interventions on diabetes in a prospective study of healthy women, using the parametric g-formula. Using data from the Nurses' Health Study, we followed 76,402 women from 1984 to 2008. We estimated the risk of type 2 diabetes under eight hypothetical interventions: quitting smoking, losing weight by 5% every 2 years if overweight/obese, exercising at least 30 minutes a day, eating less than three servings a week of red meat, eating at least two servings a day of whole grain, drinking two or more cups of coffee a day, drinking five or more grams of alcohol a day, and drinking less than one serving of soda a week. The 24-year risk of diabetes was 9.6% under no intervention and 4.3% when all interventions were imposed (55% lower risk [95% confidence interval = 47 to 63%]). The most effective interventions were weight loss (24% lower risk), physical activity (19%), and moderate alcohol use (19%). Overweight/obese women would benefit the most, with 10.8 percentage point reduction in 24-year risk of diabetes. The validity of these estimates relies on the absence of unmeasured confounding, measurement error, and model misspecification. A combination of dietary and nondietary lifestyle modifications, begun in midlife or later in relatively healthy women, could have prevented at least half of the cases of type 2 diabetes in this cohort of U.S. women.


Biohybrid robotics: From the nanoscale to the macroscale.
Biohybrid robotics is a field in which biological entities are combined with artificial materials in order to obtain improved performance or features that are difficult to mimic with hand-made materials. Three main level of integration can be envisioned depending on the complexity of the biological entity, ranging from the nanoscale to the macroscale. At the nanoscale, enzymes that catalyze biocompatible reactions can be used as power sources for self-propelled nanoparticles of different geometries and compositions, obtaining rather interesting active matter systems that acquire importance in the biomedical field as drug delivery systems. At the microscale, single enzymes are substituted by complete cells, such as bacteria or spermatozoa, whose self-propelling capabilities can be used to transport cargo and can also be used as drug delivery systems, for in vitro fertilization practices or for biofilm removal. Finally, at the macroscale, the combinations of millions of cells forming tissues can be used to power biorobotic devices or bioactuators by using muscle cells. Both cardiac and skeletal muscle tissue have been part of remarkable examples of untethered biorobots that can crawl or swim due to the contractions of the tissue and current developments aim at the integration of several types of tissue to obtain more realistic biomimetic devices, which could lead to the next generation of hybrid robotics. Tethered bioactuators, however, result in excellent candidates for tissue models for drug screening purposes or the study of muscle myopathies due to their three-dimensional architecture. This article is categorized under: Therapeutic Approaches and Drug Discovery > Emerging Technologies Nanotechnology Approaches to Biology > Nanoscale Systems in Biology.