Red wine: A drink to your heart.
Mortality and morbidity are still high in cardiovascular disease (CVD). Myocardial ischemia reperfusion injury leading to myocardial infarction is one of the most frequent causes of the death in humans. Atherosclerosis and generation of reactive oxygen species through oxidative stress is the major risk factor for CVD. From the literature collection, it has been identified that moderate consumption of red wine helps in preventing CVD through several mechanisms, including increasing the high-density lipoprotein cholesterol plasma levels, decreasing platelet aggregation, by antioxidant effects, and by restoration of endothelial function. The aim of this review is to discuss the accumulating evidence that suggests that red wine possesses a diverse range of biological actions and may be beneficial in the prevention of CVD.


A preliminary controlled study to determine whether whole-plant cannabis extracts can improve intractable neurogenic symptoms.
To determine whether plant-derived cannabis medicinal extracts (CME) can alleviate neurogenic symptoms unresponsive to standard treatment, and to quantify adverse effects. A consecutive series of double-blind, randomized, placebo-controlled single-patient cross-over trials with two-week treatment periods. Patients attended as outpatients, but took the CME at home. Twenty-four patients with multiple sclerosis (18), spinal cord injury (4), brachial plexus damage (1), and limb amputation due to neurofibromatosis (1). Whole-plant extracts of delta-9-tetrahydrocannabinol (THC), cannabidiol (CBD), 1:1 CBD:THC, or matched placebo were self-administered by sublingual spray at doses determined by titration against symptom relief or unwanted effects within the range of 2.5-120 mg/24 hours. Measures used: Patients recorded symptom, well-being and intoxication scores on a daily basis using visual analogue scales. At the end of each two-week period an observer rated severity and frequency of symptoms on numerical rating scales, administered standard measures of disability (Barthel Index), mood and cognition, and recorded adverse events. Pain relief associated with both THC and CBD was significantly superior to placebo. Impaired bladder control, muscle spasms and spasticity were improved by CME in some patients with these symptoms. Three patients had transient hypotension and intoxication with rapid initial dosing of THC-containing CME. Cannabis medicinal extracts can improve neurogenic symptoms unresponsive to standard treatments. Unwanted effects are predictable and generally well tolerated. Larger scale studies are warranted to confirm these findings.


Prevalence of central vein stenosis following catheterization in patients with end-stage renal disease.
To determine prevalence of central vein stenosis following catheterization with double-lumen temporary catheters, we performed color Doppler sonography in 100 consecutive patients. We detected central vein stenosis in 18 cases; 11 patients in subclavian vein (SCV), 4 patients in internal jugular vein (IJV) and SCV, 2 patients in SCV and brachiocephalic vein, and 2 patients in IJV stenosis. There were statistical difference between groups with and without stenosis regarding time from discontinuation of catheters and use of aspirin (ASA). We could not find any statistical difference between these two groups regarding age, sex, duration of having chronic kidney disease (CKD), and duration of catheter remaining in place. We also found that there was a high proportion of stenosis in patients who still had catheter in their veins (15 from 44 patients, 34%) in comparison with patients who had already the catheters removed from their veins (3 from 56 patients, 5%). We conclude that stenosis of central veins can result from long indwelling time of central catheter used for hemodialysis. Aspirin may have a protective role against stenosis.


Administration of fenoldopam in critically ill small animal patients with acute kidney injury: 28 dogs and 34 cats (2008-2012).
To describe the clinical features and outcomes of critically ill dogs and cats with acute kidney injury (AKI) receiving fenoldopam infusions compared to patients with AKI that did not receive fenoldopam. Retrospective clinical study from May 1, 2008 until June 1, 2012. Private emergency and specialty referral hospital. Client-owned dogs (28) and cats (34) with AKI that received fenoldopam compared with similar patients with AKI (30 dogs and 30 cats) that did not. None. The medical records of 62 critically ill dogs and cats with AKI that received fenoldopam were reviewed. Presenting clinical signs, physical examination findings, and primary and secondary disease processes were identified in all patients. The mean number of days on fenoldopam was 1.5 days (range 0.3-4.0 days) for dogs and 1.9 days (range 1.0-4.0 days) for cats. Eleven of 28 (39%) dogs survived to discharge and 13 of 34 (38%) of the cats survived to discharge. Of the animals in the group receiving fenoldopam that died, the majority (84%) were euthanized. Potential adverse reactions were evaluated, with hypotension being the most commonly encountered adverse effect (7% of fenoldopam group [FG] dogs and 23% of FG cats). When compared with patients with AKI that did not receive fenoldopam, no significant differences were found between the groups with regards to survival, length of hospital stay, adverse effects, or changes in creatinine, BUN, or sodium concentrations except that patients receiving fenoldopam were significantly more likely to have received other renally active medications. In this study of patients with AKI, fenoldopam administration at 0.8 μg/kg/min in dogs and 0.5 μg/kg/min in cats appeared relatively safe but was not associated with improvement in survival to discharge, length of hospital stay, or improvement in renal biochemical parameters when compared to patients with AKI not receiving fenoldopam.


A mutation in X-linked inhibitor of apoptosis (G466X) leads to memory inflation of Epstein-Barr virus-specific T cells.
Mutations in the X-linked inhibitor of apoptosis (XIAP) gene have been associated with XLP-like disease, including recurrent Epstein-Barr virus (EBV)-related haemophagocytic lymphohystiocytosis (HLH), but the immunopathogenic bases of EBV-related disease in XIAP deficiency is unknown. We present the first analysis of EBV-specific T cell responses in functional XIAP deficiency. In a family of patients with a novel mutation in XIAP (G466X) leading to a late-truncated protein and varying clinical features, we identified gradual hypogammaglobulinaemia and large expansions of T cell subsets, including a prominent CD4(+) CD8(+) population. Extensive ex-vivo analyses showed that the expanded T cell subsets were dominated by EBV-specific cells with conserved cytotoxic, proliferative and interferon (IFN)-γ secretion capacity. The EBV load in blood fluctuated and was occasionally very high, indicating that the XIAP(G466X) mutation could impact upon EBV latency. XIAP deficiency may unravel a new immunopathogenic mechanism in EBV-associated disease.


Stem cells in the hair follicle bulge contribute to wound repair but not to homeostasis of the epidermis.
The discovery of long-lived epithelial stem cells in the bulge region of the hair follicle led to the hypothesis that epidermal renewal and epidermal repair after wounding both depend on these cells. To determine whether bulge cells are necessary for epidermal renewal, here we have ablated these cells by targeting them with a suicide gene encoding herpes simplex virus thymidine kinase (HSV-TK) using a Keratin 1-15 (Krt1-15) promoter. We show that ablation leads to complete loss of hair follicles but survival of the epidermis. Through fate-mapping experiments, we find that stem cells in the hair follicle bulge do not normally contribute cells to the epidermis which is organized into epidermal proliferative units, as previously predicted. After epidermal injury, however, cells from the bulge are recruited into the epidermis and migrate in a linear manner toward the center of the wound, ultimately forming a marked radial pattern. Notably, although the bulge-derived cells acquire an epidermal phenotype, most are eliminated from the epidermis over several weeks, indicating that bulge stem cells respond rapidly to epidermal wounding by generating short-lived 'transient amplifying' cells responsible for acute wound repair. Our findings have implications for both gene therapy and developing treatments for wounds because it will be necessary to consider epidermal and hair follicle stem cells as distinct populations.


Bioaccumulation of polystyrene nanoplastics and their effect on the toxicity of Au ions in zebrafish embryos.
As nano- and micro-sized plastics accumulate in the environment and the food chain of animals, including humans, it is imperative to assess the effects of nanoplastics in living organisms in a systematic manner, especially because of their ability to adsorb potential toxicants such as pollutants, heavy metals, and organic macromolecules that coexist in the environment. Using the zebrafish embryo as an animal model, we investigated the bioaccumulation and in vivo toxicity of polystyrene (PS) nanoplastics individually or in combination with the Au ion. We showed that smaller PS nanoplastics readily penetrated the chorion and developing embryos and accumulated throughout the whole body, mostly in lipid-rich regions such as in yolk lipids. We also showed that PS nanoplastics induced only marginal effects on the survival, hatching rate, developmental abnormalities, and cell death of zebrafish embryos but that these effects were synergistically exacerbated by the Au ion in a dose- and size-dependent manner. Such exacerbation of toxicity was well correlated with the production of reactive oxygen species and the pro-inflammatory responses synergized by the presence of PS, supporting the combined toxicity of PS and Au ions. The synergistic effect of PS on toxicity appeared to relate to mitochondrial damage as determined by ultrastructural analysis. Taken together, the effects of PS nanoplastics were marginal but could be a trigger for exacerbating the toxicity induced by other toxicants such as metal ions.


Putative Secondary Structure at 5'UTR as a Potential Antiviral Target against SARS-CoV-2.
SARS-CoV-2 is an enveloped positive-sense single-stranded RNA coronavirus that causes COVID-19, of which the current outbreak has resulted in a high number of cases and fatalities throughout the world, even vaccine doses are being administered. The aim of this work was to scan the SARS-CoV-2 genome in search for therapeutic targets. We found a sequence in the 5'UTR (NC\_045512:74-130), consisting of a typical heptamer next to a structured region that may cause ribosomal frameshifting. The potential biological value of this region is relevant through its low similarity with other viruses, including coronaviruses related to SARS-CoV, and its high sequence conservation within multiple SARS-CoV-2 isolates. We have predicted the secondary structure of the region by means of different bioinformatic tools. We have suggested a most probable secondary structure to proceed with a 3D reconstruction of the structured segment. Finally, we carried out virtual docking on the 3D structure to look for a binding site and then for drug ligands from a database of lead compounds. Several molecules that could be probably administered as oral drugs show promising binding affinity within the structured region, and so it could be possible interfere its potential regulatory role.


First Report of Ageratum yellow vein China virus Infecting Zinnia elegans in Vietnam.
Whitefly-transmitted geminiviruses belong to the Geminiviridae family, which includes a group of plant DNA viruses that occurs in tropical and subtropical regions worldwide and caused severe leaf curl diseases on many crops. In July 2011, symptoms of mild yellowing and curling of leaves were observed on Zinnia elegans in Hue City, Vietnam. To identify possible begomoviruses, two samples with typical symptoms (Z1 and Z2) were collected and total DNA was extracted using PlantGen DNA Kit (CoWin Biotech Co., China). Using begomovirus-specific degenerate primer pair AV494 and AC1048 to amply geminivirus capsid protein gene (3), a 586-bp DNA fragment was amplified from both collected samples. The PCR products from Z1 and Z2 were cloned and sequenced. The partial sequences from two isolates had 96.8% similarity to each other and most closely related to Ageratum yellow vein China virus (AYVCNV) (98% identity to GenBank Accession Nos. AM940137.1 and AJ849916.1). Using two degenerate primer pairs bego1 (5'-ACMGGCCYATGWAYMGGAAG-3')/bego6 (5'-ACNGGDAARACRATGTGGGC-3') and bego2 (5'-CTTDATRTTBTCRTCCATCC-3')/bego3 (5'-CCWGGATTGCANARSAMGAT-3') to amplify full-length viral genome from Z1 (1). The complete DNA-A sequence from Z1 were determined to be 2,741 nucleotides (JX911332), showed high levels of sequence identity (89% [JN809812] to 98% [AJ849916.1 and AJ558120]) with AYVCNV and shared 98% nucleotide sequence identity with AYVCNV-G68 (AJ849916.1) and AYVCNV-G13 (AJ558120). No DNA-B component nor DNA-β could be detected using primers described previously (4). Zinnia leaf curl virus, Ageratum enation virus, Tobacco leaf curl virus, and Alternanthera yellow vein virus have been reported to be associated with Zinnia leaf curl disease (2). To our knowledge, this is the first report of AYVCNV infecting Z. elegans. AYVCNV was first reported in A. conyzoides on Hainan Island, China, and could infect L. esculentum (Solanaceae), A. conyzoides (Asteraceae), or tobacco (Solanaceae). Our results further corroborate that AYVCNV can infect Asteraceae plants. Therefore, this virus may pose a serious threat to many plants. References: (1) L. Lin et al. Plant Prot. 37:44, 2011. (2) P. Naveen et al. J. Biotechnol. Bioinform. 2:6, 2012. (3) S. D. Wyatt et al. Phytopathology 86:1288, 1996. (4) X. P. Zhou et al. J. Gen. Virol. 84:237, 2003.


Multiple enzymatic activities of ParB/Srx superfamily mediate sexual conflict among conjugative plasmids.
Conjugative plasmids are typically locked in intergenomic and sexual conflicts with co-resident rivals, whose translocation they block using fertility inhibition factors (FINs). We describe here the first crystal structure of an enigmatic FIN Osa deployed by the proteobacterial plasmid pSa. Osa contains a catalytically active version of the ParB/Sulfiredoxin fold with both ATPase and DNase activity, the latter being regulated by an ATP-dependent switch. Using the Agrobacterium tumefaciens VirB/D4 type IV secretion system (T4SS), a relative of the conjugative T4SS, we demonstrate that catalytically active Osa blocks T-DNA transfer into plants. With a partially reconstituted T4SS in vitro, we show that Osa degrades T-DNA in the T-DNA-VirD2 complex before its translocation. Further, we present evidence for conservation and interplay between ATPase and DNase activities throughout the ParB/Sulfiredoxin fold, using other members of the family, namely P1 ParB and RK2 KorB, which have general functional implications across diverse biological contexts.


Validity, reliability and applicability of Portuguese versions of sedation-agitation scales among critically ill patients.
Sedation scales are used to guide sedation protocols in intensive care units (ICUs). However, no sedation scale in Portuguese has ever been evaluated. The aim of this study was to evaluate the validity and reliability of Portuguese translations of four sedation-agitation scales, among critically ill patients: Glasgow Coma Score, Ramsay, Richmond Agitation-Sedation Scale (RASS) and Sedation-Agitation Scale (SAS). Validation study in two mixed ICUs of a university hospital. All scales were applied to 29 patients by four different critical care team members (nurse, physiotherapist, senior critical care physician and critical care resident). We tested each scale for interrater reliability and for validity, by correlations between them. Interrater agreement was measured using weighted kappa (k) and correlations used Spearman's test. 136 observations were made on 29 patients. All scales had at least substantial agreement (weighted k 0.68-0.90). RASS (weighted k 0.82-0.87) and SAS (weighted k 0.83-0.90) had the best agreement. All scales had a good and significant correlation with each other. All scales demonstrated good interrater reliability and were comparable. RASS and SAS showed the best correlations and the best agreement results in all professional categories. All these characteristics make RASS and SAS good scales for use at the bedside, to evaluate sedation-agitation among critically ill patients in terms of validity, reliability and applicability.


Explaining socioeconomic differences in sickness absence: the Whitehall II Study.
To describe and explain the socioeconomic gradient in sickness absence. Analysis of questionnaire and sickness absence data collected from the first phase of the Whitehall II study. Grade of employment was used as a measure of socioeconomic status. 20 civil service departments in London. 6900 male and 3414 female civil servants aged 35-55 years. Rates of short spells (< or = 7 days) and long spells (> 7 days) of sickness absence. A strong inverse relation between grade of employment and sickness absence was evident. Men in the lowest grade had rates of short and long spells of absence 6.1 (95% confidence interval 5.3 to 6.9) and 6.1 (4.8 to 7.9) times higher than those in the highest grade. For women the corresponding rate ratios were 3.0 (2.3 to 3.9) and 4.2 (2.5 to 6.8) respectively. Several risk factors were identified, including health related behaviours (smoking and frequent alcohol consumption), work characteristics (low levels of control, variety and use of skills, work pace, and support at work), low levels of job satisfaction, and adverse social circumstances outside work (financial difficulties and negative support). These risk factors accounted for about one third of the grade differences in sickness absence. Large grade differences in sickness absence parallel socioeconomic differences in morbidity and mortality found in other studies. Identified risk factors accounted for a small proportion of the grade differences in sickness absence. More accurate measurement of the risk factors may explain some of the remaining differences in sickness absence but other factors, as yet unrecognised, are likely to be important.


Seedling Stage Resistance of Iranian Bread Wheat Germplasm to Race Ug99 of Puccinia graminis f. sp. tritici.
Following emergence of Ug99, the new virulent race of Puccinia graminis f. sp. tritici in Africa, a global effort for identification and utilization of new sources of Ug99-resistant germplasm has been undertaken. In this study, we conducted replicated experiments to evaluate the resistance of Iranian wheat germplasm to the TTKSK lineage of the Ug99 race of P. graminis f. sp. tritici. We also evaluated for presence of stem rust resistance genes (i.e., Sr2, Sr24, Sr26, Sr38, Sr39, Sr31, and Sr1RSAmigo) in wheat cultivars and breeding lines widely cultivated in Iran. Our phenotyping data revealed high levels of susceptibility to Ug99 in Iranian bread wheat germplasm. Our genotyping data revealed that Iranian cultivars do not carry Sr24, Sr26, or Sr1RSAmigo. Only a few salt-tolerant cultivars and breeding lines tested positively for Sr2, Sr31, Sr38, or Sr39 markers. In conclusion, the genetic basis for resistance to Ug99 in Iranian wheat cultivars was found to be vulnerable. Acquiring knowledge about existing resistance genes and haplotypes in wheat cultivars and breeding lines will help breeders, cereal pathologists, and policy makers to select and pyramid effective stem rust resistance genes.


Use of reactive tracers to determine ambient OH radical concentrations: application within the indoor environment.
The hydroxyl radical (OH) plays a key role in determining indoor air quality. However, its highly reactive nature and low concentration indoors impede direct analysis. This paper describes the techniques used to indirectly quantify indoor OH, including the development of a new method based on the instantaneous release of chemical tracers into the air. This method was used to detect ambient OH in two indoor seminar rooms following tracer detection by gas chromatography-mass spectrometry (GCMS). The results from these tests add to the small number of experiments that have measured indoor OH which are discussed with regard to future directions within air quality research.


Dynamic vapor microextraction of ignitable liquid from casework containers.
Dynamic vapor microextraction (DVME) is a headspace concentration method that can be used to collect ignitable liquid (IL) from fire debris onto chilled adsorbent capillaries. Unlike passive headspace concentration onto activated carbon strips (ACSs) that must be eluted with a toxic solvent (carbon disulfide), DVME employs a relatively benign solvent (acetone) to recover the adsorbed IL residue, and each headspace collection is monitored for breakthrough. Here, for the first time, we extend DVME to casework containers while exploring a realistic range of oven temperatures and collection volumes. We investigated metal cans sealed with friction lids (container 1), metal cans sealed within polymer bags (container 2), and glass jars sealed with two-piece lids (container 3). Without additional containment, container 1 was found to leak so excessively that flow through the capillary was unreliable. Therefore, for containers 2 and 3 only, we determined the total number of target compounds collected from 50% weathered gasoline for oven temperatures from 54 °C to 96 °C and collection volumes from 47 standard cubic centimeters (scc) to 90 scc. Only high-volatility species with retention times (tR)< n-decane on a non-polar column were recovered from polymer bags, whereas headspace concentration from glass jars led to the recovery of target compounds across the entire volatility range. DVME at 90 °C from 2-mL containers showed that the presence of polymer bag material leads to IL vapor losses, particularly for low-volatility species with tR> n-decane. DVME was strongly influenced by the casework container, whereas oven temperature and collection volume had a minor influence for the IL samples explored here.


Parameter learning from stochastic teachers and stochastic compulsive liars.
This paper considers a general learning problem akin to the field of learning automata (LA) in which the learning mechanism attempts to learn from a stochastic teacher or a stochastic compulsive liar. More specifically, unlike the traditional LA model in which LA attempts to learn the optimal action offered by the Environment (also here called the "Oracle"), this paper considers the problem of the learning mechanism (robot, an LA, or in general, an algorithm) attempting to learn a "parameter" within a closed interval. The problem is modeled as follows: The learning mechanism is trying to locate an unknown point on a real interval by interacting with a stochastic Environment through a series of informed guesses. For each guess, the Environment essentially informs the mechanism, possibly erroneously (i.e., with probability p), which way it should move to reach the unknown point. When the probability of a correct response is p > 0.5, the Environment is said to be informative, and thus the case of learning from a stochastic teacher. When this probability p < 0.5, the Environment is deemed deceptive, and is called a stochastic compulsive liar. This paper describes a novel learning strategy by which the unknown parameter can be learned in both environments. These results are the first reported results, which are applicable to the latter scenario. The most significant contribution of this paper is that the proposed scheme is shown to operate equally well, even when the learning mechanism is unaware of whether the Environment ("Oracle") is informative or deceptive. The learning strategy proposed herein, called CPL-AdS, partitions the search interval into d subintervals, evaluates the location of the unknown point with respect to these subintervals using fast-converging E-optimal LRI LA, and prunes the search space in each iteration by eliminating at least one partition. The CPL-AdS algorithm is shown to provably converge to the unknown point with an arbitrary degree of accuracy with probability as close to unity as desired. Comprehensive experimental results confirm the fast and accurate convergence of the search for a wide range of values for the Environment's feedback accuracy parameter p, and thus has numerous potential applications.


Anti-tick saliva antibody: a biologic marker of tick exposure that is a risk factor for Lyme disease seropositivity.
Anti-tick saliva antibody (ATSA) has potential as a biologic marker of exposure to tick bites. In 1989, we conducted a cross-sectional study of 304 outdoor workers in Monmouth County, New Jersey, to evaluate associations between self-reported tick exposure, ATSA status, and Lyme disease antibody status. ATSA levels 1) were correlated with an index of tick exposure on the basis of three self-reported measures of tick exposure and outdoor hours worked per week (p = 0.01); 2) were consistently higher in pet owners compared with persons without pets (p = 0.03); and 3) when examined by duration since last tick bite, peaked at 3-5 weeks after tick bite and then declined (p = 0.06). ATSA levels dichotomized at the 75th percentile (approximately two standard deviations above the mean ATSA optical density of 25 subjects who denied recent tick exposure) were associated with self-reported tick exposure: adjusted odds ratios for high ATSA levels were 4.2 (95% confidence interval (CI) 0.9-18.9) for moderate (versus none) tick exposure and 5.8 (95% CI 1.2-27.2) for high (versus none) tick exposure. Finally, high ATSA levels were associated with Lyme disease seropositivity, with an adjusted odds ratio of 3.2 (95% CI 1.3-7.6). The data suggest that ATSA is a biologic marker of tick exposure that is a risk factor for Lyme disease seropositivity.


Scorpion toxins targeting Kv1.3 channels: insights into immunosuppression.
Scorpion venoms are natural sources of molecules that have, in addition to their toxic function, potential therapeutic applications. In this source the neurotoxins can be found especially those that act on potassium channels. Potassium channels are responsible for maintaining the membrane potential in the excitable cells, especially the voltage-dependent potassium channels (Kv), including Kv1.3 channels. These channels (Kv1.3) are expressed by various types of tissues and cells, being part of several physiological processes. However, the major studies of Kv1.3 are performed on T cells due its importance on autoimmune diseases. Scorpion toxins capable of acting on potassium channels (KTx), mainly on Kv1.3 channels, have gained a prominent role for their possible ability to control inflammatory autoimmune diseases. Some of these toxins have already left bench trials and are being evaluated in clinical trials, presenting great therapeutic potential. Thus, scorpion toxins are important natural molecules that should not be overlooked in the treatment of autoimmune and other diseases.


Impacts of a prekindergarten program on children's mathematics, language, literacy, executive function, and emotional skills.
Publicly funded prekindergarten programs have achieved small-to-large impacts on children's cognitive outcomes. The current study examined the impact of a prekindergarten program that implemented a coaching system and consistent literacy, language, and mathematics curricula on these and other nontargeted, essential components of school readiness, such as executive functioning. Participants included 2,018 four and five-year-old children. Findings indicated that the program had moderate-to-large impacts on children's language, literacy, numeracy and mathematics skills, and small impacts on children's executive functioning and a measure of emotion recognition. Some impacts were considerably larger for some subgroups. For urban public school districts, results inform important programmatic decisions. For policy makers, results confirm that prekindergarten programs can improve educationally vital outcomes for children in meaningful, important ways.


Rumen Protozoa Play a Significant Role in Fungal Predation and Plant Carbohydrate Breakdown.
The rumen protozoa, alongside fungi, comprise the eukaryotic portion of the rumen microbiome. Rumen protozoa may account for up to 50% of biomass, yet their role in this ecosystem remains unclear. Early experiments inferred a role in carbohydrate and protein metabolism, but due to their close association with bacteria, definitively attributing these functions to the protozoa was challenging. The advent of 'omic technologies has created opportunities to broaden our understanding of the rumen protozoa. This study aimed to utilize these methods to further our understanding of the role that protozoa play in the rumen in terms of their metabolic capacities, and in doing so, contribute valuable sequence data to reduce the chance of mis or under-representation of the rumen protozoa in meta'omic datasets. Rumen protozoa were isolated and purified using glucose-based sedimentation and differential centrifugation, extracted RNA was Poly(A) fraction enriched and DNase treated before use in a phage-based, cDNA metatranscriptomic library. Biochemical activity testing of the phage library showed 6 putatively positive plaques in response to carboxymethyl cellulose agar (indicative of cellulose activity), and no positive results for tributyrin (indicative of esterase/lipase activity) or egg yolk agar (indicative of proteolysis). Direct sequencing of the cDNA was also conducted using the Illumina HiSeq 2500. The metatranscriptome identified a wealth of carbohydrate-active enzymes which accounted for 8% of total reads. The most highly expressed carbohydrate-active enzymes were glycosyl hydrolases 5 and 11, polysaccharide lyases and deacetylases, xylanases and enzymes active against pectin, mannan and chitin; the latter likely used to digest rumen fungi which contain a chitin-rich cell membrane. Codon usage analysis of expressed genes also showed evidence of horizontal gene transfer, suggesting that many of these enzymes were acquired from the rumen bacteria in an evolutionary response to the carbohydrate-rich environment of the rumen. This study provides evidence of the significant contribution that the protozoa make to carbohydrate breakdown in the rumen, potentially using horizontally acquired genes, and highlights their predatory capacity.


Analysis of maize (Zea mays L.) seedling roots with the high-throughput image analysis tool ARIA (Automatic Root Image Analysis).
The maize root system is crucial for plant establishment as well as water and nutrient uptake. There is substantial genetic and phenotypic variation for root architecture, which gives opportunity for selection. Root traits, however, have not been used as selection criterion mainly due to the difficulty in measuring them, as well as their quantitative mode of inheritance. Seedling root traits offer an opportunity to study multiple individuals and to enable repeated measurements per year as compared to adult root phenotyping. We developed a new software framework to capture various traits from a single image of seedling roots. This framework is based on the mathematical notion of converting images of roots into an equivalent graph. This allows automated querying of multiple traits simply as graph operations. This framework is furthermore extendable to 3D tomography image data. In order to evaluate this tool, a subset of the 384 inbred lines from the Ames panel, for which extensive genotype by sequencing data are available, was investigated. A genome wide association study was applied to this panel for two traits, Total Root Length and Total Surface Area, captured from seedling root images from WinRhizo Pro 9.0 and the current framework (called ARIA) for comparison using 135,311 single nucleotide polymorphism markers. The trait Total Root Length was found to have significant SNPs in similar regions of the genome when analyzed by both programs. This high-throughput trait capture software system allows for large phenotyping experiments and can help to establish relationships between developmental stages between seedling and adult traits in the future.


Evaluation of blood flow measurements with microspheres and rubidium--an experimental study in rats.
The microsphere method has been widely used for blood flow measurements in normal and tumour tissues. The microsphere method was evaluated for repeated measurements of cardiac output and regional blood flow in anesthetised rats and in anesthetised rats given noradrenalin and thereby having altered haemodynamics with special emphasis on liver blood flow. Comparing the microsphere method with the soluble indicator method (86Rubidium) gave equal cardiac output values. The liver blood flow was lower and the spleen blood flow was higher with the microsphere method. Two microsphere injections at 10 min intervals were performed on anesthetised rats. In one group 817 +/- 10(3) microspheres were injected each time, in a second group 436 +/- 10(3) and in a third noradrenalin was added and then 430 +/- 10(3) microspheres injected twice. There was good reproducibility for cardiac output and for most organ and tissue blood flows between first and second microsphere injection. No influence on arterial liver blood flow was seen. A blood pressure fall and a decreased heart rate was registered after the first injection in the group given 817 x 10(3) spheres. There was also a blood pressure fall in the group given noradrenalin after the first microsphere injection. The microsphere method with two injections of 436 x 10(3) microspheres seems adequate to use in arterial blood flow studies of the liver and simultaneous cardiac output measurements.


Circadian gene variants influence sleep and the sleep electroencephalogram in humans.
The sleep electroencephalogram (EEG) is highly heritable in humans and yet little is known about the genetic basis of inter-individual differences in sleep architecture. The aim of this study was to identify associations between candidate circadian gene variants and the polysomnogram, recorded under highly controlled laboratory conditions during a baseline, overnight, 8 h sleep opportunity. A candidate gene approach was employed to analyze single-nucleotide polymorphisms from five circadian-related genes in a two-phase analysis of 84 healthy young adults (28 F; 23.21 ± 2.97 years) of European ancestry. A common variant in Period2 (PER2) was associated with 20 min less slow-wave sleep (SWS) in carriers of the minor allele than in noncarriers, representing a 22% reduction in SWS duration. Moreover, spectral analysis in a subset of participants (n = 37) showed the same PER2 polymorphism was associated with reduced EEG power density in the low delta range (0.25-1.0 Hz) during non-REM sleep and lower slow-wave activity (0.75-4.5 Hz) in the early part of the sleep episode. These results indicate the involvement of PER2 in the homeostatic process of sleep. Additionally, a rare variant in Melatonin Receptor 1B was associated with longer REM sleep latency, with minor allele carriers exhibiting an average of 65 min (87%) longer latency from sleep onset to REM sleep, compared to noncarriers. These findings suggest that circadian-related genes can modulate sleep architecture and the sleep EEG, including specific parameters previously implicated in the homeostatic regulation of sleep.


Nivolumab for Relapsed/Refractory Classic Hodgkin Lymphoma After Failure of Autologous Hematopoietic Cell Transplantation: Extended Follow-Up of the Multicohort Single-Arm Phase II CheckMate 205 Trial.
Purpose Genetic alterations causing overexpression of programmed death-1 ligands are near universal in classic Hodgkin lymphoma (cHL). Nivolumab, a programmed death-1 checkpoint inhibitor, demonstrated efficacy in relapsed/refractory cHL after autologous hematopoietic cell transplantation (auto-HCT) in initial analyses of one of three cohorts from the CheckMate 205 study of nivolumab for cHL. Here, we assess safety and efficacy after extended follow-up of all three cohorts. Methods This multicenter, single-arm, phase II study enrolled patients with relapsed/refractory cHL after auto-HCT treatment failure into cohorts by treatment history: brentuximab vedotin (BV)-naïve (cohort A), BV received after auto-HCT (cohort B), and BV received before and/or after auto-HCT (cohort C). All patients received nivolumab 3 mg/kg every 2 weeks until disease progression/unacceptable toxicity. The primary end point was objective response rate per independent radiology review committee. Results Overall, 243 patients were treated; 63 in cohort A, 80 in cohort B, and 100 in cohort C. After a median follow-up of 18 months, 40% continued to receive treatment. The objective response rate was 69% (95% CI, 63% to 75%) overall and 65% to 73% in each cohort. Overall, the median duration of response was 16.6 months (95% CI, 13.2 to 20.3 months), and median progression-free survival was 14.7 months (95% CI, 11.3 to 18.5 months). Of 70 patients treated past conventional disease progression, 61% of those evaluable had stable or further reduced target tumor burdens. The most common grade 3 to 4 drug-related adverse events were lipase increases (5%), neutropenia (3%), and ALT increases (3%). Twenty-nine deaths occurred; none were considered treatment related. Conclusion With extended follow-up, responses to nivolumab were frequent and durable. Nivolumab seems to be associated with a favorable safety profile and long-term benefits across a broad spectrum of patients with relapsed/refractory cHL.


TCF7L2 regulates late events in insulin secretion from pancreatic islet beta-cells.
Polymorphisms in the human TCF7L2 gene are associated with reduced insulin secretion and an increased risk of type 2 diabetes. However, the mechanisms by which TCF7L2 affect insulin secretion are still unclear. We define the effects of TCF7L2 expression level on mature beta-cell function and suggest a potential mechanism for its actions. TCF7L2 expression in rodent islets and beta-cell lines was altered using RNAi or adenoviral transduction. Beta-cell gene profiles were measured by quantitative real-time PCR and the effects on intracellular signaling and exocytosis by live cell imaging, electron microscopy, and patch clamp electrophysiology. Reducing TCF7L2 expression levels by RNAi decreased glucose- but not KCl-induced insulin secretion. The glucose-induced increments in both ATP/ADP ratio and cytosolic free Ca2+ concentration ([Ca2+]i) were increased compared with controls. Overexpression of TCF7L2 exerted minor inhibitory effects on glucose-regulated changes in [Ca2+]i and insulin release. Gene expression profiling in TCF7L2-silenced cells revealed increased levels of mRNA encoding syntaxin 1A but decreased Munc18-1 and ZnT8 mRNA. Whereas the number of morphologically docked vesicles was unchanged by TCF7L2 suppression, secretory granule movement increased and capacitance changes decreased, indicative of defective vesicle fusion. TCF7L2 is involved in maintaining expression of beta-cell genes regulating secretory granule fusion. Defective insulin exocytosis may thus underlie increased diabetes incidence in carriers of the at-risk TCF7L2 alleles.


Alexithymia and its relationship to dissociation in patients with panic disorder.
Conditions that impede the regulation of emotional arousal, such as alexithymia and dissociation, may underlie panic attacks. This study aimed to evaluate the relationship between alexithymia and dissociation in patients with panic disorder (PD). We assessed 95 PD outpatients with regard to alexithymia (20-item Toronto Alexithymia Scale), dissociation (Dissociation Experience Scale), and overall psychological distress (Symptom Checklist 90-Revised, Global Severity Index). Regression analyses revealed a positive correlation between alexithymia and dissociation, even when the Global Severity Index was controlled for. A specific link was observed between "difficulty in identifying feelings" and "depersonalization/derealization." Patients who showed the pathological form of dissociation had higher levels of alexithymia, with particular regard to "difficulty in identifying feelings" and, to a smaller extent, "difficulty in describing feelings." These results support a strong relationship between alexithymia and dissociation in patients with PD. Assessing alexithymia and dissociation at the outset of therapy may be helpful for individualized therapy planning.


Aldehyde dehydrogenase (ALDH) - a promising new candidate for use in preclinical and clinical selection of pluripotent very small embryonic-like stem cells (VSEL SCs) of high long-term repopulating hematopoietic potential.
In the field of stem cells (SCs) biology there is great need for a universal marker that can effectively identify the rare populations of SCs in order to characterize and isolate them for research and therapeutic purposes. In line with this idea, the measurement of aldehyde dehydrogenase (ALDH) activity shows promising potential as a common marker for the identification of SCs. Recently discovered very small embryonic-like stem cells (VSEL SCs) have a pluripotent nature and high long-term repopulating hematopoietic potential. The aim of the present study was to determine whether VSEL SCs isolated from umbilical cord blood (CB) and different murine organs express high ALDH activity. To address this issue, 12 CB units were analyzed by employing flow cytometry to detect the following populations: (i) CXCR4(+)/Lin(-)/CD45(-)/ALDH(high), (ii) CD34(+)/Lin(-)/CD45(-)/ALDH(high) and (iii) CD133(+)/Lin(-)/CD45(-)/ALDH(high). Additionally, selected organs from 16 5-week-old female inbred C57BL/6J mice and 16 7-month-old female C57BL/6J mice were analyzed for detection of the following populations: (i) Sca-1(+)/Lin(-)/CD45(-) and (ii) Sca-1(+)/Lin(-)/CD45(+). All these populations were assessed for ALDH activity. We found that CB contains VSEL SCs with high ALDH activity. We also observed that ALDH(high) cells constitute a modest percentage of VSEL SCs present in selected murine organs. We demonstrated that CB and adult murine organs possess a subpopulation of ALDH(high) VSEL SCs. Above all, the observed high level of ALDH activity can be considered a functional marker of organ-derived pluripotent SCs and allows for simple, efficient isolation of cells with primitive features for their utility in targeted cell therapies.


Correlation of depressive symptoms and olfactory dysfunction in patients on hemodialysis.
A possible link between depression and olfactory dysfunction has been suggested in the literature, in research projects using the olfactory bulbectomy model. In human studies using a syndrome-oriented approach, such an association has not been reported consistently. The aim of the study was to test the association of olfactory dysfunction with depression using a symptom-oriented approach.Paients and methods: Twenty eight end-stage renal failure patients took part in this project. The patients olfactory identification ability was tested with the University of Pennsylvania Smell Identification Test (UPSIT). Immediately before olfactory testing, the subjects completed the Zung self-rating scale, which provides data on symptoms of depression in this group of patients. The mean value of the number of mistakes made in the olfactory identification ability (UPSIT test) by the total sample was 14.0±4.5, with a range 6-22. Half of the symptoms seem to bear an influence on the olfactory identification performance. Patients experiencing decreased libido and dissatisfaction exhibited significantly reduced olfactory function, as contrasted to those not experiencing these symptoms. The above results remain practically unaltered even after taking into account such probable confounding factors as age, sex, olfactory detection threshold and duration of illness. These findings support previous evidence indicating that olfactory dysfunction may be related to specific depressive symptoms in humans. The present findings also suggest that the symptom-oriented approach is an effective research tool for the elucidation of such clinical issues. The need for further research in this field is pointed out.


Expansion of a Population of Large Monocytes (Atypical Monocytes) in Peripheral Blood of Patients with Acute Exacerbations of Chronic Obstructive Pulmonary Diseases.
Acute exacerbation of chronic obstructive pulmonary disease (AECOPD) is closely associated with airway inflammation including monocytes, lymphocytes, and neutrophils. Monocytes play an essential role in the pathogenesis of chronic obstructive pulmonary disease (COPD). To elucidate the association of circulating monocyte alteration with AECOPD, we analyzed monocyte subpopulation in the peripheral blood of 16 healthy volunteers and 22 AECOPD patients at the stages of admission and remission after clinical therapy. We found a dramatic increase of a previously unreported population of large size circulating atypical monocytes (A Mo) in AECOPD patients, characterized by higher forward scatter and lower side scatter values than the typical monocytes (T Mo) which were observed predominantly in healthy individuals. Further analysis showed that A Mo expressed higher levels of CD16, intercellular adhesion molecule 1 (ICAM-1), and chemotactic protein-1 receptor-2 (CCR2) than T Mo. In contrast, the expression of class II antigen (HLA-DR) by A Mo was lower than T Mo. More importantly, we observed that the percentage of circulating A Mo among total monocytes correlated with the length of hospital stay (time to remission) and disease duration. The data suggest that circulating A Mo might have the potential to serve as a biomarker in the diagnosis and prognosis of AECOPD.


Women's Fertility Status Alters Other Women's Jealousy and Mate Guarding.
Across three studies, we tested the hypothesis that women exhibit greater jealousy and mate guarding toward women who are in the high (vs. low) fertility phase of their cycle. Women who imagined their partner with a woman pictured at high fertility reported more jealousy than women who imagined their partner with a woman pictured at low fertility (Studies 1 and 2). A meta-analysis across studies manipulating fertility status of the pictured woman found a significant effect of fertility status on both jealousy and mate guarding. Women with attractive partners viewed fertile-phase women as less trustworthy, which led to increased mate guarding (Study 2). In Study 3, the closer women were to peak fertility, the more instances they reported of other women acting jealously and mate guarding toward them. These studies provide evidence that women selectively exhibit jealousy and mate guarding toward women who are near peak fertility.


Minor components with smooth muscle relaxing properties from scented myrrh (Commiphora guidotti).
All sesquiterpenes present in a sample of scented myrrh were isolated and characterised. Seven compounds, with cadinane, guaiane, oplopane, and eudesmane skeletons, were obtained, of which two are new and two are reported from a natural source for the first time. The major component, T-cadinol, has previously been shown to possess smooth muscle-relaxing properties, and the major purpose of the investigation was to compare the effects of the minor and more polar sesquiterpenes with that of T-cadinol in the rat aorta. Like T-cadinol, the minor sesquiterpenes are more efficient in reducing K(+)-induced contractions than those induced by the alpha-adrenoceptor agonist phenylephrine, however, they were all less potent than T-cadinol.


Confronting uncertainty and missing values in environmental value transfer as applied to species conservation.
The nonuse (or passive) value of nature is important but time-consuming and costly to quantify with direct surveys. In the absence of estimates of these values, there will likely be less investment in conservation actions that generate substantial nonuse benefits, such as conservation of native species. To help overcome decisions about the allocation of conservation dollars that reflect the lack of estimates of nonuse values, these values can be estimated indirectly by environmental value transfer (EVT). EVT uses existing data or information from a study site such that the estimated monetary value of an environmental good is transferred to another location or policy site. A major challenge in the use of EVT is the uncertainty about the sign and size of the error (i.e., the percentage by which transferred value exceeds the actual value) that results from transferring direct estimates of nonuse values from a study to a policy site, the site where the value is transferred. An EVT is most useful if the decision-making framework does not require highly accurate information and when the conservation decision is constrained by time and financial resources. To account for uncertainty in the decision-making process, a decision heuristic that guides the decision process and illustrates the possible decision branches, can be followed. To account for the uncertainty associated with the transfer of values from one site to another, we developed a risk and simulation approach that uses Monte Carlo simulations to evaluate the net benefits of conservation investments and takes into account different possible distributions of transfer error. This method does not reduce transfer error, but it provides a way to account for the effect of transfer error in conservation decision making. Our risk and simulation approach and decision-based framework on when to use EVT offer better-informed decision making in conservation.


Prediction of Survival After Partial Hepatectomy Using a Physiologically Based Pharmacokinetic Model of Indocyanine Green Liver Function Tests.
The evaluation of hepatic function and functional capacity of the liver are essential tasks in hepatology as well as in hepatobiliary surgery. Indocyanine green (ICG) is a widely applied test compound that is used in clinical routine to evaluate hepatic function. Important questions for the functional evaluation with ICG in the context of hepatectomy are how liver disease such as cirrhosis alters ICG elimination, and if postoperative survival can be predicted from preoperative ICG measurements. Within this work a physiologically based pharmacokinetic (PBPK) model of ICG was developed and applied to the prediction of the effects of a liver resection under various degrees of cirrhosis. For the parametrization of the computational model and validation of model predictions a database of ICG pharmacokinetic data was established. The model was applied (i) to study the effect of liver cirrhosis and liver resection on ICG pharmacokinetics; and (ii) to evaluate the model-based prediction of postoperative ICG-R15 (retention ratio 15 min after administration) as a measure for postoperative outcome. Key results are the accurate prediction of changes in ICG pharmacokinetics caused by liver cirrhosis and postoperative changes of ICG-elimination after liver resection, as validated with a wide range of data sets. Based on the PBPK model, individual survival after liver resection could be classified, demonstrating its potential value as a clinical tool.


Immobilization of cellulose fibrils on solid substrates for cellulase-binding studies through quantitative fluorescence microscopy.
Cellulases, enzymes capable of depolymerizing cellulose polymers into fermentable sugars, are essential components in the production of bioethanol from lignocellulosic materials. Given the importance of these enzymes to the evolving biofuel industry considerable research effort is focused on understanding the interaction between cellulases and cellulose fibrils. This manuscript presents a method that addresses challenges that must be overcome in order to study such interactions through high-resolution fluorescence microscopy. First, it is shown that cellulose can be immobilized on solid substrates through a polymer lift-off technique. The immobilized cellulose aggregates present characteristic morphologies influenced by the patterned feature size used to immobilize it. Thus, through a variety of pattern sizes, cellulose can be immobilized in the form of cellulose particles, cellulose mats or individual cellulose fibrils. Second, it is shown that both cellulose and Thermobifida fusca cellulases Cel5A, Cel6B, and Cel9A can be fluorescently tagged and that the labeling does not inhibit the capability of these cellulases to depolymerize cellulose. The combination of the immobilization technique together with fluorescence labeling yields a system that can be used to study cellulose-cellulase interactions with spatial and temporal resolution not available through more conventional techniques which measure ensemble averages. It is shown that with such a system, the kinetics of cellulase binding onto cellulose fibrils and mats can be followed through sequences of fluorescence images. The intensity from the images can then be used to reconstruct binding curves for the cellulases studied. It was found that the complexity of cellulose morphology has a large impact on the binding curve characteristics, with binding curves for individual cellulose fibrils closely following a binding saturation model and binding curves for cellulose mats and particles deviating from it. The behavior observed is interpreted as the effect pore and interstice penetration play in cellulase binding to the accessible surface of cellulose aggregates. These results validate our method for immobilizing nanoscale cellulose fibrils and fibril aggregates on solid supports and lay the foundation for future studies on cellulase-cellulose interactions.


Optimal delineation of single C-tactile and C-nociceptive afferents in humans by latency slowing.
C-mechanoreceptors in humans comprise a population of unmyelinated afferents exhibiting a wide range of mechanical sensitivities. C-mechanoreceptors are putatively divided into those signaling gentle touch (C-tactile afferents, CTs) and nociception (C-mechanosensitive nociceptors, CMs), giving rise to positive and negative affect, respectively. We sought to distinguish, compare, and contrast the properties of a population of human C-mechanoreceptors to see how fundamental the divisions between these putative subpopulations are. We used microneurography to record from individual afferents in humans and applied electrical and mechanical stimulation to their receptive fields. We show that C-mechanoreceptors can be distinguished unequivocally into two putative populations, comprising CTs and CMs, by electrically evoked spike latency changes (slowing). After both natural mechanical stimulation and repetitive electrical stimulation there was markedly less latency slowing in CTs compared with CMs. Electrical receptive field stimulation, which bypasses the receptor end organ, was most effective in classifying C-mechanoreceptors, as responses to mechanical receptive field stimulation overlapped somewhat, which may lead to misclassification. Furthermore, we report a subclass of low-threshold CM responding to gentle mechanical stimulation and a potential subclass of CT afferent displaying burst firing. We show that substantial differences exist in the mechanisms governing axonal conduction between CTs and CMs. We provide clear electrophysiological "signatures" (extent of latency slowing) that can be used in unequivocally identifying populations of C-mechanoreceptors in single-unit and multiunit microneurography studies and in translational animal research into affective touch. Additionally, these differential mechanisms may be pharmacologically targetable for separate modulation of positive and negative affective touch information.NEW & NOTEWORTHY Human skin encodes a plethora of touch interactions, and affective tactile information is primarily signaled by slowly conducting C-mechanoreceptive afferents. We show that electrical stimulation of low-threshold C-tactile afferents produces markedly different patterns of activity compared with high-threshold C-mechanoreceptive nociceptors, although the populations overlap in their responses to mechanical stimulation. This fundamental distinction demonstrates a divergence in affective touch signaling from the first stage of sensory processing, having implications for the processing of interpersonal touch.


Corticosteroids in critical COVID-19: Are all corticosteroids equal?
The hyperinflammation seen as part of a dysregulated immune response to SARS-CoV-2 in its most severe form leads to acute respiratory distress syndrome (ARDS), multiorgan failure and death. Corticosteroid therapy targets this hyperinflammation, otherwise known as a cytokine storm. It is the only therapeutic agent to date with a mortality benefit, with clear guidelines from national and international health authorities guiding its use. Objectives. To compare severity-of-illness indices, survival, length of intensive care unit (ICU) stay and potential ICU complications in patients treated with different corticosteroid regimens (high-dose hydrocortisone, high-dose methylprednisolone and lower-dose dexamethasone). Methods. In this single-centre descriptive retrospective observational study of a cohort of patients with severe COVID-19 admitted to a COVID-dedicated ICU, we compared patients treated with the three different corticosteroid regimens. Results. In 242 cases we could not demonstrate any statistically or clinically significant difference in the outcome of patients with critical COVID-19 treated with high-dose intravenous hydrocortisone (n=88) or methylprednisolone (n=46) compared with a relatively lower dose of dexamethasone (n=108). The survival rates were 38.6%, 39.1% and 33.3%, respectively (p=0.68). Patients treated with methylprednisolone tended to have a shorter length of ICU stay (median (interquartile range) 6 (4 - 10), 4 (2 - 8) and 5 (2 - 8) days; p=0.015) and fewer episodes of nosocomial sepsis (47.7%, 32.6% and 48.1%; p=0.01). Conclusions. Hydrocortisone or methylprednisolone can be given as an alternative to dexamethasone in the management of critical COVID-19, and this is a feasible alternative, especially in resource-constrained settings.


Adaptive sugar sensors in hypothalamic feeding circuits.
Brain glucose sensing is critical for healthy energy balance, but how appropriate neurocircuits encode both small changes and large background values of glucose levels is unknown. Here, we report several features of hypothalamic orexin neurons, cells essential for normal wakefulness and feeding: (i) A distinct group of orexin neurons exhibits only transient inhibitory responses to sustained rises in sugar levels; (ii) this sensing strategy involves time-dependent recovery from inhibition via adaptive closure of leak-like K(+) channels; (iii) combining transient and sustained glucosensing allows orexin cell firing to maintain sensitivity to small fluctuations in glucose levels while simultaneously encoding a large range of baseline glucose concentrations. These data provide insights into how vital behavioral orchestrators sense key features of the internal environment while sustaining a basic activity tone required for the stability of consciousness.


Prevalence of cardiometabolic risk factors and metabolic syndrome in obese Kuwaiti adolescents.
Childhood and adolescent obesity is associated with insulin resistance, abnormal glucose metabolism, hypertension, dyslipidemia, inflammation, liver disease, and compromised vascular function. The purpose of this pilot study was to determine the prevalence of cardiometabolic risk factor abnormalities and metabolic syndrome (MetS) in a sample of obese Kuwaiti adolescents, as prevalence data might be helpful in improving engagement with obesity treatment in future. Eighty obese Kuwaiti adolescents (40 males) with a mean (standard deviation) age of 12.3 years (1.1 years) participated in the present study. All participants had a detailed clinical examination and anthropometry, blood pressure taken, and assessment of fasting levels of C-reactive protein, intracellular adhesion molecule, interleukin-6, fasting blood glucose, insulin, liver function tests (alanine aminotransferase, aspartate aminotransferase, gamma glutamyltransferase), lipid profile (cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides), insulin resistance by homeostasis model assessment, and adiponectin. MetS was assessed using two recognized criteria modified for use in younger individuals. The cardiometabolic risk factors with highest prevalence of abnormal values included aspartate aminotransferase (88.7% of the sample) and insulin resistance by homeostasis model assessment (67.5%), intracellular adhesion molecule (66.5%), fasting insulin (43.5%), C-reactive protein (42.5%), low-density lipoprotein cholesterol (35.0%), total cholesterol (33.5%), and systolic blood pressure (30.0%). Of all participants, 96.3% (77/80) had at least one impaired cardiometabolic risk factor as well as obesity. Prevalence of MetS was 21.3% according to the International Diabetes Federation definition and 30% using the Third Adult Treatment Panel definition. The present study suggests that obese Kuwaiti adolescents have multiple cardiometabolic risk factor abnormalities. Future studies are needed to test the benefits of intervention in this high-risk group. They also suggest that prevention of obesity in children and adults should be a major public health goal in Kuwait.


Nicotinic acid activates the capsaicin receptor TRPV1: Potential mechanism for cutaneous flushing.
Nicotinic acid (also known as niacin or vitamin B3), widely used to treat dyslipidemias, represents an effective and safe means to reduce the risk of mortality from cardiovascular disease. Nonetheless, a substantial fraction of patients discontinue treatment because of a strong side effect of cutaneous vasodilation, commonly termed flushing. In the present study, we tested the hypothesis that nicotinic acid causes flushing partially by activating the capsaicin receptor TRPV1, a polymodal cellular sensor that mediates the flushing response on consumption of spicy food. We observed that the nicotinic acid-induced increase in blood flow was substantially reduced in Trpv1(-/-) knockout mice, indicating involvement of the channel in flushing response. Using exogenously expressed TRPV1, we confirmed that nicotinic acid at submillimolar to millimolar concentrations directly and potently activates TRPV1 from the intracellular side. Binding of nicotinic acid to TRPV1 lowers its activation threshold for heat, causing channel opening at physiological temperatures. The activation of TRPV1 by voltage or ligands (capsaicin and 2-aminoethoxydiphenyl borate) is also potentiated by nicotinic acid. We further demonstrated that nicotinic acid does not compete directly with capsaicin but may activate TRPV1 through the 2-aminoethoxydiphenyl borate activation pathway. Using live-cell fluorescence imaging, we observed that nicotinic acid can quickly enter the cell through a transporter-mediated pathway to activate TRPV1. Direct activation of TRPV1 by nicotinic acid may lead to cutaneous vasodilation that contributes to flushing, suggesting a potential novel pathway to inhibit flushing and to improve compliance.


Structure and activation mechanism of the human liver-type glutaminase GLS2.
Cancer cells exhibit an altered metabolic phenotype, consuming higher levels of the amino acid glutamine. This metabolic reprogramming depends on increased mitochondrial glutaminase activity to convert glutamine to glutamate, an essential precursor for bioenergetic and biosynthetic processes in cells. Mammals encode the kidney-type (GLS) and liver-type (GLS2) glutaminase isozymes. GLS is overexpressed in cancer and associated with enhanced malignancy. On the other hand, GLS2 is either a tumor suppressor or an oncogene, depending on the tumor type. The GLS structure and activation mechanism are well known, while the structural determinants for GLS2 activation remain elusive. Here, we describe the structure of the human glutaminase domain of GLS2, followed by the functional characterization of the residues critical for its activity. Increasing concentrations of GLS2 lead to tetramer stabilization, a process enhanced by phosphate. In GLS2, the so-called "lid loop" is in a rigid open conformation, which may be related to its higher affinity for phosphate and lower affinity for glutamine; hence, it has lower glutaminase activity than GLS. The lower affinity of GLS2 for glutamine is also related to its less electropositive catalytic site than GLS, as indicated by a Thr225Lys substitution within the catalytic site decreasing the GLS2 glutamine concentration corresponding to half-maximal velocity (K0.5). Finally, we show that the Lys253Ala substitution (corresponding to the Lys320Ala in the GLS "activation" loop, formerly known as the "gating" loop) renders a highly active protein in stable tetrameric form. We conclude that the "activation" loop, a known target for GLS inhibition, may also be a drug target for GLS2.


Consumption of Whole-Grain Bread and Risk of Colorectal Cancer among Norwegian Women (the NOWAC Study).
There is evidence that consumption of foods containing dietary fiber decreases the risk of colorectal cancer (CRC). Whole grains contain dietary fiber, as well as a range of micronutrients and bioactive compounds, but the association between the consumption of whole grains and the risk of CRC remains less studied. The aim of the present study was to investigate the association between whole-grain bread consumption and CRC incidence among Norwegian women, using data from a prospective cohort study (the Norwegian Women and Cancer Study). Dietary intake was estimated from the food-frequency questionnaires of 78,254 women in the cohort (median age: 55 years), and these women were then followed up for CRC incidence. During the 9 years of median follow-up, 795 women were diagnosed with CRC (316 proximal, 193 distal, 218 rectal). Associations between whole-grain bread consumption and the risk of CRC (including colorectal subsites) were investigated using Cox proportional hazards regression models. When compared to the low consumption group, the hazard ratio for CRC was 0.89 (95% confidence interval (CI): 0.72-1.09) for the high consumption group and 0.86 (95% CI: 0.72-1.02) for the medium consumption group in a multivariable model. Overall, no association between whole-grain bread consumption and CRC was found.


Evolutionary tradeoffs can select against nitrogen fixation and thereby maintain nitrogen limitation.
Symbiotic nitrogen (N) fixing trees are absent from old-growth temperate and boreal ecosystems, even though many of these are N-limited. To explore mechanisms that could select against N fixation in N-limited, old-growth ecosystems, we developed a simple resource-based evolutionary model of N fixation. When there are no costs of N fixation, increasing amounts of N fixation will be selected for until N no longer limits production. However, tradeoffs between N fixation and plant mortality or turnover, plant uptake of available soil N, or N use efficiency (NUE) can select against N fixation in N-limited ecosystems and can thereby maintain N limitation indefinitely (provided that there are losses of plant-unavailable N). Three key traits influence the threshold that determines how large these tradeoffs must be to select against N fixation. A low NUE, high mortality (or turnover) rate and low losses of plant-unavailable N all increase the likelihood that N fixation will be selected against, and a preliminary examination of published data on these parameters shows that these mechanisms, particularly the tradeoff with NUE, are quite feasible in some systems. Although these results are promising, a better characterization of these parameters in multiple ecosystems is necessary to determine whether these mechanisms explain the lack of symbiotic N fixers-and thus the maintenance of N limitation-in old-growth forests.


Metabolic syndrome and risks of colon and rectal cancer: the European prospective investigation into cancer and nutrition study.
Metabolic syndrome (MetS) is purportedly related to risk of developing colorectal cancer; however, the association of MetS, as defined according to recent international criteria, and colorectal cancer has not been yet evaluated. In particular, it remains unclear to what extent the MetS components individually account for such an association. We addressed these issues in a nested case-control study that included 1,093 incident cases matched (1:1) to controls by using incidence density sampling. Conditional logistic regression was used to estimate relative risks (RR) and 95% CIs. MetS was defined according to the criteria of the National Cholesterol Education Program/Adult Treatment Panel III (NCEP/ATPIII), the International Diabetes Federation (IDF), and the 2009 harmonized definition. Among individual components, abdominal obesity (RR = 1.51; 95% CI: 1.16-1.96) was associated with colon cancer, whereas abnormal glucose metabolism was associated with both colon (RR = 2.05; 95% CI: 1.57-2.68) and rectal cancer (RR = 2.07; 95% CI: 1.45-2.96). MetS, as defined by each of the definitions, was similarly associated with colon cancer (e.g., RR = 1.91; 95% CI: 1.47-2.42 for MetS by NCEP/ATPIII), whereas MetS by NCEP/ATPIII, but not IDF or harmonized definition, was associated with rectal cancer (RR = 1.45; 95% CI: 1.02-2.06). Overall, these associations were stronger in women than in men. However, the association between MetS and colorectal cancer was accounted for by abdominal obesity and abnormal glucose metabolism such that MetS did not provide risk information beyond these components (likelihood ratio test P = 0.10 for MetS by NCEP/ATPIII). These data suggest that simple assessment of abnormal glucose metabolism and/or abdominal obesity to identify individuals at colorectal cancer risk may have higher clinical utility than applying more complex MetS definitions.


Pump efficiency improvement of a C-band tunable fiber laser using optical circulator and tunable fiber gratings.
We propose and demonstrate a tunable fiber laser based on an optical circulator (OC) and two tunable fiber Bragg gratings (TFBGs). The OC acts as a pump power router to improve the pumping efficiency, and a 4% increase in overall conversion efficiency has been observed. The combined tuning spectra range of two TFBGs could cover the entire C-band spectrum from 1530 to 1560 nm. Stable laser output power above 10 dBm is obtained using 1.9 m of erbium-doped fiber and TFBGs with 50% reflectivity. With power equalization by using variable optical attenuators, the power variation is less than 0.1 dB in the whole C band with narrow linewidth of 0.05 nm. A signal-to-noise ratio of 60 dB and a continuous tuning resolution of 0.5 nm have been achieved. The TFBG-based tunable fiber laser can be a promising light source for WDM transmission and fiber sensor applications.


Prolonged Treatment of Peanut-Allergic Mice with Bortezomib Significantly Reduces Serum Anti-Peanut IgE but Does Not Affect Allergic Symptoms.
Anti-peanut immunoglobulin E (anti-Pn IgE) can persist throughout life, suggesting that this condition could be maintained by long-lived antibody-secreting cells (ASCs). To determine the role of long-lived ASCs, peanut-allergic mice underwent prolonged treatment with the proteasome inhibitor, bortezomib (Bz). Intravenous Bz was given twice weekly for 21 weeks to peanut-allergic mice. During treatment, serum anti-Pn IgE was measured, and the mice were rechallenged at the end of treatment. Cell populations were measured, and Pn-specific IgG, total IgG, and total IgE ASCs were enumerated in the bone marrow (BM) and spleen (SPL). Prolonged treatment with Bz significantly reduced serum anti-Pn IgE and IgG1 but did not affect symptoms following challenge with Pn, even in mice with undetectable serum anti-Pn IgE. Numbers of CD138+ cells were significantly reduced in the BM but were unaffected in the SPL. Unexpectedly, Bz did not affect numbers of Pn-specific IgG, total IgG, or total IgE ASCs in either the BM or SPL. Cells that maintain long-lived serum anti-Pn IgE are sensitive to Bz. However, prolonged depletion of serum Pn-specific IgE does not result in a decrease of symptoms following challenge with Pn.


Cystathionine γ Lyase Sulfhydrates the RNA Binding Protein Human Antigen R to Preserve Endothelial Cell Function and Delay Atherogenesis.
Hydrogen sulfide (H2S), generated by cystathionine γ lyase (CSE), is an important endogenous regulator of vascular function. The aim of the present study was to investigate the control and consequences of CSE activity in endothelial cells under physiological and proatherogenic conditions. Endothelial cell CSE knockout mice were generated, and lung endothelial cells were studied in vitro (gene expression, protein sulfhydration, and monocyte adhesion). Mice were crossed onto the apolipoprotein E-deficient background, and atherogenesis (partial carotid artery ligation) was monitored over 21 days. CSE expression, H2S bioavailability, and amino acid profiling were also performed with human material. The endothelial cell-specific deletion of CSE selectively increased the expression of CD62E and elevated monocyte adherence in the absence of an inflammatory stimulus. Mechanistically, CD62E mRNA was more stable in endothelial cells from CSE-deficient mice, an effect attributed to the attenuated sulfhydration and dimerization of the RNA-binding protein human antigen R. CSE expression was upregulated in mice after partial carotid artery ligation and in atheromas from human subjects. Despite the increase in CSE protein, circulating and intraplaque H2S levels were reduced, a phenomenon that could be attributed to the serine phosphorylation (on Ser377) and inhibition of the enzyme, most likely resulting from increased interleukin-1β. Consistent with the loss of H2S, human antigen R sulfhydration was attenuated in atherosclerosis and resulted in the stabilization of human antigen R-target mRNAs, for example, CD62E and cathepsin S, both of which are linked to endothelial cell activation and atherosclerosis. The deletion of CSE from endothelial cells was associated with the accelerated development of endothelial dysfunction and atherosclerosis, effects that were reversed on treatment with a polysulfide donor. Finally, in mice and humans, plasma levels of the CSE substrate l-cystathionine negatively correlated with vascular reactivity and H2S levels, indicating its potential use as a biomarker for vascular disease. The constitutive S-sulfhydration of human antigen R (on Cys13) by CSE-derived H2S prevents its homodimerization and activity, which attenuates the expression of target proteins such as CD62E and cathepsin S. However, as a consequence of vascular inflammation, the beneficial actions of CSE-derived H2S are lost owing to the phosphorylation and inhibition of the enzyme.


Lateral Hypothalamic GABAergic Neurons Encode Reward Predictions that Are Relayed to the Ventral Tegmental Area to Regulate Learning.
Eating is a learned process. Our desires for specific foods arise through experience. Both electrical stimulation and optogenetic studies have shown that increased activity in the lateral hypothalamus (LH) promotes feeding. Current dogma is that these effects reflect a role for LH neurons in the control of the core motivation to feed, and their activity comes under control of forebrain regions to elicit learned food-motivated behaviors. However, these effects could also reflect the storage of associative information about the cues leading to food in LH itself. Here, we present data from several studies that are consistent with a role for LH in learning. In the first experiment, we use a novel GAD-Cre rat to show that optogenetic inhibition of LH γ-aminobutyric acid (GABA) neurons restricted to cue presentation disrupts the rats' ability to learn that a cue predicts food without affecting subsequent food consumption. In the second experiment, we show that this manipulation also disrupts the ability of a cue to promote food seeking after learning. Finally, we show that inhibition of the terminals of the LH GABA neurons in ventral-tegmental area (VTA) facilitates learning about reward-paired cues. These results suggest that the LH GABA neurons are critical for storing and later disseminating information about reward-predictive cues.


Aerobic granular sludge in an SBR-system treating wastewater rich in particulate matter.
Aerobic granular sludge was successfully cultivated in a lab-scale SBR-system treating malting wastewater with a high content of particulate organic matter (0.9 gTSS/L). At an organic loading rate (CODtotal) of 3.4 kg/(m3 x d) an average removal efficiency of 50% in CODtotal and 80% in CODdissolved was achieved. Fractionation of the COD by means of particle size showed that particles with a diameter less than 25-50 microm could be removed at 80% efficiency, whereas particles bigger than 50 microm were only removed at 40% efficiency. Tracer experiments revealed a dense sessile protozoa population covering the granules. The protozoa appeared to be responsible for primary particle uptake from the wastewater.


Implicating a role for immune recognition of self in tumor rejection: passive immunization against the brown locus protein.
The immune system can recognize differentiation antigens that are selectively expressed on malignant cells and their normal cell counterparts. However, it is uncertain whether immunity to differentiation antigens can effectively lead to tumor rejection. The mouse brown locus protein, gp75 or tyrosinase-related protein 1, is a melanocyte differentiation antigen expressed by melanomas and normal melanocytes. The gp75 antigen is recognized by autoantibodies and autoreactive T cells in persons with melanoma. To model autoimmunity against a melanocyte differentiation antigen, mouse antibodies against gp75 were passively transferred into tumor-bearing mice. Passive immunization with a mouse monoclonal antibody against gp75 induced protection and rejection of both subcutaneous tumors and lung metastases in syngeneic C57BL/6 mice, including established tumors. Passive immunity produced coat color alterations but only in regenerating hairs. This system provides a model for autoimmune vitiligo and shows that immune responses to melanocyte differentiation antigens can influence mouse coat color. Immune recognition of a melanocyte differentiation antigen can reject tumors, providing a basis for targeting tissue autoantigens expressed on cancer.


Physiological implications of the contrasting modulation of the activities of the epsilon- and zeta-isoforms of diacylglycerol kinase.
We have shown that the requirement of the epsilon-isoform of diacylglycerol kinase for diacylglycerols containing arachidonic acid is specific for this substrate and cannot be replaced by the presence of an arachidonoyl group in other places in the membrane; rather, it has to be present on the substrate itself. In addition, we demonstrate that the increased activity shown toward 1-stearoyl-2-arachidonoylglycerol by the epsilon-isoform of diacylglycerol kinase is not a consequence of altered membrane physical properties but is rather a specific interaction with the arachidonoyl group. We have also compared the modulation of the activity of the epsilon-isoform of diacylglycerol kinase with that of the zeta-isoform with regard to some of the intermediates involved in phosphatidylinositol cycling. One of the products of the hydrolysis of phosphatidylinositol diphosphate is diacylglycerol enriched in arachidonic acid. The activity of the epsilon-isoform is known to be specific for this form of diacylglycerol. We show that in contrast, the activity of the zeta-isoform is lower against 1-stearoyl-2-arachidonoylglycerol compared with dioleoylglycerol. We demonstrate that addition of phosphatidylserine, as well as other anionic phospholipids including L-alpha-phosphatidylinositol 4,5-bisphosphate, strongly inhibits the epsilon-isoform, but these anionic lipids increase the activity of the zeta-isoform. Addition of Ca(2+), which is released from internal stores as a consequence of phosphatidylinositol cycling, promotes the activity of the epsilon-isoform of this enzyme but has little effect on the zeta-isoform. The contrasting conditions required for maximal activity of these two isoforms of diacylglycerol kinase, as well as their different substrate specificity, suggest that they have different physiological roles in signal transduction.


Polymeric Based Therapeutic Delivery Systems Prepared Using Electrohydrodynamic Processes.
The development of therapeutic dosage (e.g. pharmaceutical) systems is an ongoing process which, in recent times has incorporated several emerging disciplines and themes at timely intervals. While the concepts surrounding dosage forms have developed and evolved, many polymeric excipients remain as the preferred choice of materials over existing counterparts, serving functions as matrix materials, coatings and providing other specific functional properties (e.g. adhesion, controlled release and mechanical properties). There have been, however, developments in the deployment of synthetic polymeric materials (e.g. polycaprolactone, poly lactic co-glycolic acid) when compared to naturally occurring materials (e.g. lactose, gelatin). Advances in pharmaceutical process technologies have also provided novel engineering platforms to develop a host of exciting structure based materials ranging from the nanometer to the macro scales. Some of these structure enabling technologies include spray drying, super critical processing, microfluidics and even wet chemical methods. More recently electrohydrodynamic (EHDA) engineering methods have emerged as robust technologies offering potential to fabricate a plethora of generic structures (e.g. particles, fibres, bubbles and pre-determined patterns) on a broad scale range. This review focuses on key developments using various EHDA technologies for the pharmaceutical and biomaterial remits when selecting synthetic and/or naturally occurring polymers as pharmaceutical (and therapeutic) excipients. In addition, the underlying EHDA process principles are discussed along with key parameters and variables (both materials and engineering). EHDA technologies are operational at ambient conditions and recent developments have also demonstrated their viability for large scale production. These are promising technologies which have potential in established (e.g. films, dressings and microparticles) and emerging scientific themes (e.g. nanomedicines and tissue engineering).


Electrochemical CO2 reduction on nanostructured metal electrodes: fact or defect?
Electrochemical CO2 reduction has received an increased amount of interest in the last decade as a promising avenue for storing renewable electricity in chemical bonds. Despite considerable progress on catalyst performance using nanostructured electrodes, the sensitivity of the reaction to process conditions has led to debate on the origin of the activity and high selectivity. Additionally, this raises questions on the transferability of the performance and knowledge to other electrochemical systems. At its core, the discrepancy is primarily a result of the highly porous nature of nanostructured electrodes, which are vulnerable to both mass transport effects and structural changes during the electrolysis. Both effects are not straightforward to identify and difficult to decouple. Despite the susceptibility of nanostructured electrodes to mass transfer limitations, we highlight that nanostructured silver electrodes exhibit considerably higher activity when normalized to the electrochemically active surface in contrast to gold and copper electrodes. Alongside, we provide a discussion on how active surface area and thickness of the catalytic layer itself can influence the onset potential, selectivity, stability, activity and mass transfer inside and outside of the three dimensional catalyst layer. Key parameters and potential solutions are highlighted to decouple mass transfer effects from the measured activity in electrochemical cells utilizing CO2 saturated aqueous solutions.


Bridging the gap between science and decision making.
All decisions, whether they are personal, public, or business-related, are based on the decision maker's beliefs and values. Science can and should help decision makers by shaping their beliefs. Unfortunately, science is not easily accessible to decision makers, and scientists often do not understand decision makers' information needs. This article presents a framework for bridging the gap between science and decision making and illustrates it with two examples. The first example is a personal health decision. It shows how a formal representation of the beliefs and values can reflect scientific inputs by a physician to combine with the values held by the decision maker to inform a medical choice. The second example is a public policy decision about managing a potential environmental hazard. It illustrates how controversial beliefs can be reflected as uncertainties and informed by science to make better decisions. Both examples use decision analysis to bridge science and decisions. The conclusions suggest that this can be a helpful process that requires skills in both science and decision making.


Impact of natural gas extraction on PAH levels in ambient air.
Natural gas extraction, often referred to as "fracking," has increased rapidly in the U.S. in recent years. To address potential health impacts, passive air samplers were deployed in a rural community heavily affected by the natural gas boom. Samplers were analyzed for 62 polycyclic aromatic hydrocarbons (PAHs). Results were grouped based on distance from each sampler to the nearest active well. PAH levels were highest when samplers were closest to active wells. Additionally, PAH levels closest to natural gas activity were an order of magnitude higher than levels previously reported in rural areas. Sourcing ratios indicate that PAHs were predominantly petrogenic, suggesting that elevated PAH levels were influenced by direct releases from the earth. Quantitative human health risk assessment estimated the excess lifetime cancer risks associated with exposure to the measured PAHs. Closest to active wells, the risk estimated for maximum residential exposure was 2.9 in 10 000, which is above the U.S. EPA's acceptable risk level. Overall, risk estimates decreased 30% when comparing results from samplers closest to active wells to those farthest. This work suggests that natural gas extraction may be contributing significantly to PAHs in air, at levels that are relevant to human health.


RING1 proteins contribute to early proximal-distal specification of the forelimb bud by restricting Meis2 expression.
Polycomb group (PcG) proteins play a pivotal role in silencing developmental genes and help to maintain various stem and precursor cells and regulate their differentiation. PcG factors also regulate dynamic and complex regional specification, particularly in mammals, but this activity is mechanistically not well understood. In this study, we focused on proximal-distal (PD) patterning of the mouse forelimb bud to elucidate how PcG factors contribute to a regional specification process that depends on developmental signals. Depletion of the RING1 proteins RING1A (RING1) and RING1B (RNF2), which are essential components of Polycomb repressive complex 1 (PRC1), led to severe defects in forelimb formation along the PD axis. We show that preferential defects in early distal specification in Ring1A/B-deficient forelimb buds accompany failures in the repression of proximal signal circuitry bound by RING1B, including Meis1/2, and the activation of distal signal circuitry in the prospective distal region. Additional deletion of Meis2 induced partial restoration of the distal gene expression and limb formation seen in the Ring1A/B-deficient mice, suggesting a crucial role for RING1-dependent repression of Meis2 and likely also Meis1 for distal specification. We suggest that the RING1-MEIS1/2 axis is regulated by early PD signals and contributes to the initiation or maintenance of the distal signal circuitry.


Single-PPLN-assisted wavelength-/time-selective switching/dropping/swapping for 100-GHz-spaced WDM signals.
We propose an approach to implementing wavelength- and time-selective optical switching, dropping and swapping based on the sum-frequency generation (SFG) or cascaded sum- and difference-frequency generation (cSFG/DFG) in a periodically poled lithium niobate (PPLN) waveguide. Analytical solutions are derived, showing the parametric depletion effect for optical switching and the narrow-band operation due to quasi-phase matching (QPM) condition of PPLN. Using parametric depletion effect of SFG process, we demonstrate wavelength- and time-selective optical switching for ITU-grid compatible 40-Gbit/s wavelength-division multiplexed (WDM) signals with a channel spacing of 100 GHz. Less than 1-dB power penalty at a bit-error rate (BER) of 10(-9) is measured for the wavelength- and time-selective switching channel. Negligible impacts are observed on other channels of WDM signals. Using combined effects of parametric depletion and wavelength conversion of cSFG/DFG processes, we demonstrate wavelength- and time-selective optical dropping for ITU-grid compatible 100-GHz-spaced 40-Gbit/s WDM signals. Moreover, we demonstrate optical swapping between two 100-GHz-spaced 40-Gbit/s signals. The obtained theoretical and experimental results confirm single-PPLN-assisted wavelength- and time-selective optical switching, dropping and swapping for 100-GHz-spaced WDM signals, which might potentially be extended to WDM signals with narrower channel spacing.


Comparison of Sigma 1 Receptor Ligands SA4503 and PRE084 to (+)-Pentazocine in the rd10 Mouse Model of RP.
Sigma 1 receptor is a novel therapeutic target for retinal disease. Its activation, using a high-affinity, high-specificity ligand (+)-pentazocine ((+)-PTZ), rescues photoreceptor cells in the rd10 mouse model of RP. Here, we asked whether the robust retinal neuroprotective properties of (+)-PTZ are generalizable to SA4503 and PRE084, two other high-affinity sigma 1 receptor ligands. We treated 661W cells with SA4503 or PRE084. Cell viability, oxidative stress, and expression of Nrf2 and NRF2-regulated antioxidant genes (Nqo1, Cat, and Sod1) were assessed. Rd10 mice were administered SA4503 (1 mg/kg), PRE084 (0.5 mg/kg), or (+)-PTZ (0.5 mg/kg). Visual acuity, retinal architecture, and retinal electrophysiologic function were measured in vivo and retinal structure was assessed histologically. Similar to (+)-PTZ, SA4503 and PRE084 improved cell viability, attenuated oxidative stress, and increased Nrf2, Nqo1 and Cat expression. Although treatment of rd10 mice with (+)-PTZ improved visual acuity, increased outer retinal thickness, and improved photopic a- and b-wave responses compared with nontreated rd10 mice, treatment with SA4503 or PRE084 did not. The number of photoreceptor nuclei/100 µm retinal length in SA4503- and PRE084-treated rd10 mice (approximately 11/100) did not differ significantly from nontreated rd10 mice, whereas (+)-PTZ-treated mice had significantly more nuclei (approximately 22/100 µm). Cell survival and gene regulation experiments yielded similar outcomes when SA4503, PRE084, or (+)-PTZ were conducted in vitro, however neither SA4503 or PRE084 afforded in vivo protection in the severe rd10 retinopathy model comparable to (+)-PTZ. Despite all three compounds demonstrating the potential to activate sigma 1 receptor, the retinal neuroprotective properties of the three ligands differ significantly.


[Effect of pharmacotherapy on course of postoperative period after endovenous thermal ablation].
The authors assessed the effect of a micronized purified flavonoid fraction (MPFF) on the course of the postoperative period after endovenous thermal ablation (EVTA). The patients of the Study Group matching by the main studied parameters to the Control Group patients were given the MPFF according to the suggested regimen for 7 days. The obtained results were analysed by means of questionnaires (CIVIQ, VCSS, VAS) and ultrasound angioscanning. The obtained findings were statistically processed by means of the program Statistica 6.0 and reliability of the results was assessed with the help of the Student t-test. Patients of the both groups showed complete stable obliteration of the target veins. No statistically significant differences of the items of the questionnaires CIVIQ and VCSS at the beginning of the study and at the last examination were revealed, differences were noted on days 2-14 after EVTA and were not statistically significant (p>0.05). Phlebotrophic therapy in the postoperative period after EVTA helps to decrease phlebitic alterations in the coagulated vein, to improve motor activity and mental psychoemotional state of the patients.


[A new method of preparation of bovine colostral immunoglobulins for parenteral administration in calves].
A new simple method of preparation of bovine colostral immunoglobulins was described using a single step precipitation of skimmed bovine colostrum with dimethyllaurylbenzylammonium bromide (DMLBAB). This quaternary ammonium compound precipitated simultaneously nearly all colostral proteins lacking antibody activity. Bovine colostrum was collected mostly during of the first 24 hours after calving, at the latest however until 48 hours. Isolation of bovine colostral immunoglobulins proceeded as follows: one volume of skimmed colostrum containing 3-6% of immunoglobulins was slowly precipitated with the same volume of 2% water solution of DMLBAB at pH 7.9-8.1 along with continuous stirring. Turbid mixture was then heated to 43-45 degrees C and subsequently cooled to a room temperature standing overnight. Heavy precipitate sedimented down and supernatant fluid containing purified immunoglobulins was decanted and clarified by filtration. Residual DMLBAB occurring in the filtrate was removed by passage through a column of strongly acid catex, prepared in Na+ form. Purified colostral immunoglobulins were thickened to the required protein concentration by ultrafiltration. Dense retentate was clear and became an amber colour. Average yield of purified colostral immunoglobulins reached 18.8 g/litre of skimmed bovine colostrum (Tab. I). Electrophoretic purity of immunoglobulins fraction amounted to 90-95% (Fig. 1). For parenteral application in calves the above solution of immunoglobulins was subsequently adjusted to 9-11% content of protein, 0.9% of sodium chloride, pH 7.2, stabilized with 2% of aminoacetic acid and conserved with 0.015% of thiomersal. Finally, the preparation was sterilized by filtration, kept its content of immunoglobulins minimally 2 years at the temperature of storage between 2-8 degrees C and remained biologically harmless. Using the method described it was not necessary to remove casein from skimmed bovine colostrum prior to the purification of immunoglobulins. Hence the method provided a significant short cut especially in laboratory as well as pilot scale production of bovine colostral immunoglobulins bringing about a marked economic benefit.


A Computational Modeling Approach Predicts Interaction of the Antifungal Protein AFP from Aspergillus giganteus with Fungal Membranes via Its γ-Core Motif.
Fungal pathogens kill more people per year globally than malaria or tuberculosis and threaten international food security through crop destruction. New sophisticated strategies to inhibit fungal growth are thus urgently needed. Among the potential candidate molecules that strongly inhibit fungal spore germination are small cationic, cysteine-stabilized proteins of the AFP family secreted by a group of filamentous Ascomycetes. Its founding member, AFP from Aspergillus giganteus, is of particular interest since it selectively inhibits the growth of filamentous fungi without affecting the viability of mammalian, plant, or bacterial cells. AFPs are also characterized by their high efficacy and stability. Thus, AFP can serve as a lead compound for the development of novel antifungals. Notably, all members of the AFP family comprise a γ-core motif which is conserved in all antimicrobial proteins from pro- and eukaryotes and known to interfere with the integrity of cytoplasmic plasma membranes. In this study, we used classical molecular dynamics simulations combined with wet laboratory experiments and nuclear magnetic resonance (NMR) spectroscopy to characterize the structure and dynamical behavior of AFP isomers in solution and their interaction with fungal model membranes. We demonstrate that the γ-core motif of structurally conserved AFP is the key for its membrane interaction, thus verifying for the first time that the conserved γ-core motif of antimicrobial proteins is directly involved in protein-membrane interactions. Furthermore, molecular dynamic simulations suggested that AFP does not destroy the fungal membrane by pore formation but covers its surface in a well-defined manner, using a multistep mechanism to destroy the membranes integrity.IMPORTANCE Fungal pathogens pose a serious danger to human welfare since they kill more people per year than malaria or tuberculosis and are responsible for crop losses worldwide. The treatment of fungal infections is becoming more complicated as fungi develop resistances against commonly used fungicides. Therefore, discovery and development of novel antifungal agents are of utmost importance.