Intraoperative examination of sentinel lymph nodes using scrape cytology.
Background. In breast cancer, sentinel lymph node biopsy (SLNB) is widely used to assess the axilla when the nodes appear normal on palpation and ultrasonography. When the sentinel lymph nodes (SLNs) are negative, no further dissection is required. Surgical dissection or radiotherapy of the axilla is indicated for macrometastases, as well as adjuvant therapy for macrometastases and micrometastasic spread.Aim. To determine the accuracy of scrape cytology (SC) for intraoperative examination of the SLNs.Methods. SLNB was performed in 100 patients with early breast cancer in whom the axillary nodes appeared normal on clinical examination and ultrasonography. None of the patients with negative SLNs or SLNs with micrometastases only (N1mic, 0.2 - 2 mm) had further axillary dissection. Patients with SLNs containing macrometastases (N1, >2 mm) underwent axillary lymph node dissection. The results of intraoperative cytology were compared with the histopathological findings on examination of serial paraffin-embedded sections of the SLNs.Results. Intraoperative SC identified SLN metastases in 20 patients: 17 had macrometastases and 3 micrometastases. Histopathological examination of the SLNs found macrometastases in 18 patients and micrometastases in 6. SC had a sensitivity of 94% and specificity of 100% for the detection of macrometastases. For micrometastatic spread, the sensitivity and specificity were 50% and 100%, respectively.Conclusion. SC is a rapid, sensitive technique for detection of macrometastases in the SLNs of women with breast cancer. The overall accuracy in identification of any metastatic spread (N1mic, N1) to the SLNs was 85%, rising to 94% in SLNs with macrometastases.


Cranberry Proanthocyanidins Neutralize the Effects of Aggregatibacter actinomycetemcomitans Leukotoxin.
Aggregatibacter actinomycetemcomitans is a Gram-negative bacterium that has been strongly associated with localized aggressive periodontitis. The capacity of A. actinomycetemcomitans to produce a leukotoxin (LtxA) that activates pyroptosis in macrophages and induces the release of endogenous danger signals is thought to play a key role in the disease process. The aim of the present study was to investigate the effects of cranberry proanthocyanidins (PACs) on gene expression and cytotoxic activities of LtxA. We showed that cranberry PACs dose-dependently attenuate the expression of genes making up the leukotoxin operon, including ltxB and ltxC, in the two strains of A. actinomycetemcomitans tested. Cranberry PACs (≥62.5 µg/mL) protected macrophages against the cytotoxic effect of purified LtxA. Moreover, cranberry PACs reduced caspase-1 activation in LtxA-treated macrophages and consequently decreased the release of both IL-1β and IL-18, which are known as damage-associated molecular patterns (DAMPs) and contribute to the progression of periodontitis by increasing cell migration and osteoclastogenesis. In addition, cranberry PACs reduced the expression of genes encoding the P2X7 receptor and NALP3 (NACHT, LRR and PYD domains-containing protein 3), which play key roles in pore formation and cell death. Lastly, cranberry PACs blocked the binding of LtxA to macrophages and consequently reduced the LtxA-mediated cytotoxicity. In summary, the present study showed that cranberry PACs reduced LtxA gene expression in A. actinomycetemcomitans and neutralized the cytolytic and pro-inflammatory responses of human macrophages treated with LtxA. Given these properties, cranberry PACs may represent promising molecules for prevention and treatment of the aggressive form of periodontitis caused by A. actinomycetemcomitans.


Nanomaterial-based biosensors using dual transducing elements for solution phase detection.
Biosensors incorporating nanomaterials have demonstrated superior performance compared to their conventional counterparts. Most reported sensors use nanomaterials as a single transducer of signals, while biosensor designs using dual transducing elements have emerged as new approaches to further improve overall sensing performance. This review focuses on recent developments in nanomaterial-based biosensors using dual transducing elements for solution phase detection. The review begins with a brief introduction of the commonly used nanomaterial transducers suitable for designing dual element sensors, including quantum dots, metal nanoparticles, upconversion nanoparticles, graphene, graphene oxide, carbon nanotubes, and carbon nanodots. This is followed by the presentation of the four basic design principles, namely Förster Resonance Energy Transfer (FRET), Amplified Fluorescence Polarization (AFP), Bio-barcode Assay (BCA) and Chemiluminescence (CL), involving either two kinds of nanomaterials, or one nanomaterial and an organic luminescent agent (e.g. organic dyes, luminescent polymers) as dual transducers. Biomolecular and chemical analytes or biological interactions are detected by their control of the assembly and disassembly of the two transducing elements that change the distance between them, the size of the fluorophore-containing composite, or the catalytic properties of the nanomaterial transducers, among other property changes. Comparative discussions on their respective design rules and overall performances are presented afterwards. Compared with the single transducer biosensor design, such a dual-transducer configuration exhibits much enhanced flexibility and design versatility, allowing biosensors to be more specifically devised for various purposes. The review ends by highlighting some of the further development opportunities in this field.


A control switch for prothrombinase: characterization of a hirudin-like pentapeptide from the COOH terminus of factor Va heavy chain that regulates the rate and pathway for prothrombin activation.
Membrane-bound factor Xa alone catalyzes prothrombin activation following initial cleavage at Arg(271) and prethrombin 2 formation (pre2 pathway). Factor Va directs prothrombin activation by factor Xa through the meizothrombin pathway, characterized by initial cleavage at Arg(320) (meizo pathway). We have shown previously that a pentapeptide encompassing amino acid sequence 695-699 from the COOH terminus of the heavy chain of factor Va (Asp-Tyr-Asp-Tyr-Gln, DYDYQ) inhibits prothrombin activation by prothrombinase in a competitive manner with respect to substrate. To understand the mechanism of inhibition of thrombin formation by DYDYQ, we have studied prothrombin activation by gel electrophoresis. Titration of plasma-derived prothrombin activation by prothrombinase, with increasing concentrations of peptide, resulted in complete inhibition of the meizo pathway. However, thrombin formation still occurred through the pre2 pathway. These data demonstrate that the peptide preferentially inhibits initial cleavage of prothrombin by prothrombinase at Arg(320). These findings were corroborated by studying the activation of recombinant mutant prothrombin molecules rMZ-II (R155A/R284A/R271A) and rP2-II (R155A/R284A/R320A) which can be only cleaved at Arg(320) and Arg(271), respectively. Cleavage of rMZ-II by prothrombinase was completely inhibited by low concentrations of DYDYQ, whereas high concentrations of pentapeptide were required to inhibit cleavage of rP2-II. The pentapeptide also interfered with prothrombin cleavage by membrane-bound factor Xa alone in the absence of factor Va increasing the rate for cleavage at Arg(271) of plasma-derived prothrombin or rP2-II. Our data demonstrate that pentapeptide DYDYQ has opposing effects on membrane-bound factor Xa for prothrombin cleavage, depending on the incorporation of factor Va in prothrombinase.


Focus on nutrition: Home-prepared diets for dogs and cats.
Promoting health and wellness in dogs and cats is a common goal for veterinarians and pet owners alike. Over the past decade, a number of highly publicized pet food recalls, as well as a growing awareness of the role of diet in health and disease for people, have changed the way some owners approach mealtime for their pets. Many owners, and some veterinarians, now advocate feeding dogs and cats home-prepared foods (raw, cooked, or both) as the sole source of nutrition for pets and cite either perceived health benefits or a general mistrust of the pet food industry as the reason. It is important for veterinary practitioners to understand the risks and benefits of home-prepared diets, as well as the motivation behind a pet owner's decision to follow this type of feeding regimen, to ensure optimal health for their patients.


Portal inflammation is associated with advanced histological changes in alcoholic and non-alcoholic fatty liver disease.
Fatty liver disease (FLD), of either alcoholic (AFLD) or non-alcoholic (NAFLD) aetiologies, is characterised histologically by features that are lobulocentric that typically affect zone 3, in contrast with portal-based inflammation characteristics of other forms of chronic liver disease. The authors aimed to determine the prevalence and significance of portal inflammation in a cohort of adults diagnosed as having AFLD and NAFLD. From a histology database, the authors identified 160 patients with biopsy proven AFLD and 214 with NAFLD diagnosed between 1991 and 2001. All liver biopsies were reviewed by one pathologist to evaluate various histological features. More than a mild degree of portal inflammation was found in 47% of AFLD compared with 30% in NAFLD subgroup. A higher degree of portal inflammation in AFLD was associated with a higher mean corpuscular volume, gamma glutamyl transferase (GGT) and alkaline phosphatase, and a lower platelet count and albumin, whereas in NAFLD this was associated with age, presence of diabetes, hypertension, body mass index (BMI), higher fasting blood glucose, cholesterol, alanine transaminase, GGT and ferritin. On regression analysis, portal inflammation was associated with the severity of steatosis (p=0.005), presence of ballooning (p=0.030) in NAFLD and severity of fibrosis in both AFLD and NAFLD (p<0.001). Portal inflammation is a common component of histological spectrum of both AFLD and NAFLD. In both conditions portal inflammation is associated with clinical and histological features suggestive of advanced disease.


Limitations of the widely used GAM42a and BET42a probes targeting bacteria in the Gammaproteobacteria radiation.
The 23S rRNA-targeted probes GAM42a and BET42a provided equivocal results with the uncultured gammaproteobacterium 'Candidatus Competibacter phosphatis' where some cells bound GAM42a and other cells bound BET42a in fluorescence in situ hybridization (FISH) experiments. Probes GAM42a and BET42a span positions 1027-1043 in the 23S rRNA and differ from each other by one nucleotide at position 1033. Clone libraries were prepared from PCR products spanning the 16S rRNA genes, intergenic spacer region and 23S rRNA genes from two mixed cultures enriched in 'Candidatus C. phosphatis'. With individual clone inserts, the 16S rDNA portion was used to confirm the source organism as 'Candidatus C. phosphatis' and the 23S rDNA portion was used to determine the sequence of the GAM42a/BET42a probe target region. Of the 19 clones sequenced, 8 had the GAM42a probe target (T at position 1033) and 11 had G at position 1033, the only mismatch with GAM42a. However, none of the clones had the BET42a probe target (A at 1033). Non-canonical base-pairing between the 23S rRNA of 'Candidatus C. phosphatis' with G at position 1033 and GAM42a (G-A) or BET42a (G-T) is likely to explain the probing anomalies. A probe (GAM42_C1033) was optimized for use in FISH, targeting cells with G at position 1033, and was found to highlight not only some 'Candidatus C. phosphatis' cells, but also other bacteria. This demonstrates that there are bacteria in addition to 'Candidatus C. phosphatis' with the GAM42_C1033 probe target and not the BET42a or GAM42a probe target.


Crystal Structure of the Double Homeodomain of DUX4 in Complex with DNA.
Double homeobox (DUX) transcription factors are unique to eutherian mammals. DUX4 regulates expression of repetitive elements during early embryogenesis, but misexpression of DUX4 causes facioscapulohumeral muscular dystrophy (FSHD) and translocations overexpressing the DUX4 double homeodomain cause B cell leukemia. Here, we report the crystal structure of the tandem homeodomains of DUX4 bound to DNA. The homeodomains bind DNA in a head-to-head fashion, with the linker making anchoring DNA minor-groove interactions and unique protein contacts. Remarkably, despite being tandem duplicates, the DUX4 homeodomains recognize different core sequences. This results from an arginine-to-glutamate mutation, unique to primates, causing alternative positioning of a key arginine side chain in the recognition helix. Mutational studies demonstrate that this primate-specific change is responsible for the divergence in sequence recognition that likely drove coevolution of embryonically regulated repeats in primates. Our work provides a framework for understanding the endogenous function of DUX4 and its role in FSHD and cancer.


Prognostic value of routine cardiac magnetic resonance assessment of left ventricular ejection fraction and myocardial damage: an international, multicenter study.
Cardiac magnetic resonance (CMR) is considered the reference standard for assessment of left ventricular ejection fraction (LVEF) and myocardial damage. However, few studies have evaluated the relationship between CMR findings and patient outcome, and of these, most are small and none multicenter. We performed an international, multicenter study to assess the prognostic importance of routine CMR in patients with known or suspected heart disease. From 10 centers in 6 countries, consecutive patients undergoing routine CMR assessment of LVEF and myocardial damage by cine and delayed-enhancement imaging (DE-CMR), respectively, were screened for enrollment. Clinical data, CMR protocol information, and findings were collected at all sites and submitted to the data coordinating center for verification of completeness and analysis. The primary end point was all-cause mortality. A total of 1560 patients (age, 59±14 years; 70% men) were enrolled. Mean LVEF was 45±18%, and 1049 (67%) patients had hyperenhanced tissue (HE) on DE-CMR indicative of damage. During a median follow-up time of 2.4 years (interquartile range, 1.2, 2.9 years), 176 (11.3%) patients died. Patients who died were more likely to be older (P<0.0001), have coronary disease (P=0.004), have lower LVEF (P<0.0001), and have more segments with HE (P<0.0001). In multivariable analysis, age, LVEF, and number of segments with HE were independent predictors of mortality. Among patients with near-normal LVEF (≥50%), those with above-median HE (>4 segments) had reduced survival compared to patients with below- or at-median HE (P=0.02). Both LVEF and amount of myocardial damage as assessed by routine CMR are independent predictors of all-cause mortality. Even in patients with near-normal LVEF, significant damage identifies a cohort with a high risk for early mortality.


Residential modifications and decline in physical function among community-dwelling older adults.
The purpose of this study is to quantify the effect of residential modification on decreasing risk of physical function decline in 2 years. Cohort study using propensity scores method to control for baseline differences between individuals with residential modifications and those without residential modifications. Participants (N = 9,447) were from the Second Longitudinal Study on Aging, a nationally representative sample of the civilian noninstitutionalized population, aged 70 years and older in the United States at the time of baseline interview in 1994-1995. Participants self-reported residential modifications at baseline (e.g., railings, bathroom modifications). Decline in physical functioning was measured by comparing self-reported activities of daily living at baseline and at 2-year follow-up. Compared with individuals without baseline modifications, a higher proportion of those with baseline modifications were aged 85 years and older (16% vs. 10%), used special aides (36% vs. 14%), and lived alone (40% vs. 31%). Using a weighted propensity score method, we found a modest decrease in risk of decline at Wave 2 for those with baseline modifications (risk difference = 3.1%). Respondents with a baseline residential modification were less likely to experience subsequent decline in functional ability (adjusted odds ratio = 0.88, 95% confidence interval = 0.79-0.97) after adjusting for quintile of propensity score in a survey-weighted regression model. Baseline modifications may be associated with reduced risk of decline among a nationally representative sample of older community-dwelling adults. Widespread adoption of residential modifications may reduce the overall population estimates of decline.


Wheat sprout extract-induced apoptosis in human cancer cells by proteasomes modulation.
Natural occurring modulators of proteasome functionality are extensively investigated for their implication in cancer therapy. On the basis of our previous evidences both on proteasomal inhibition by monomeric polyphenols, and on the characterization of wheat sprout hydroalcoholic extract, herein we thoroughly report on a comparative study of the effect of wheat sprout extract on both normal and tumour cells. Treatment of isolated 20S proteasomes with wheat sprout extracts induced a gradual inhibition of all proteasome activities. Next, two wheat sprout extract components were separated: a polyphenol and a protein fraction. Both components exerted an in vitro inhibitory effect on proteasome activity. HeLa tumour cells and FHs 74 Int normal cells were exposed to both fractions, resulting in different rates of proteasome inhibition, with tumour cells showing a significantly higher degree of proteasome impairment and apoptosis induction. Furthermore, a decrease in proteasome activities and in cell survival of the human plasmacytoma RPMI 8226 cell line, upon the same treatments, was observed. Collectively, our results provide additional evidences supporting the possible use of natural extracts as coadjuvants in cancer treatments.


Correlation of the Ga-68-bombesin analog Ga-68-BZH3 with receptors expression in gliomas as measured by quantitative dynamic positron emission tomography (dPET) and gene arrays.
The kinetics of Ga-68-BZH3, a Ga-68-bombesin analog, was compared to molecular biological data obtained from gene arrays in seven patients with a recurrent glioma. The primary aim of this study was the correlation of receptor expression and tracer kinetics. Dynamic positron emission tomography studies were performed and the data were analyzed by a volume-of-interest technique using a two-tissue compartment model as well as a non-compartment model. Gene array data were obtained from gene array analysis of tumor tissue samples. The correlation analysis revealed a significant nonlinear correlation of r = 0.89 (p < 0.03) for k1 and BB(2) (gastrin-releasing peptide receptor). BB(1) and BB(3) were not significantly correlated with k1. vb and k3 were not significantly correlated with the expression data of the receptors on the p < 0.05 level. The parameter k1 is correlated with the expression of BB(2) based on gene array data. The quantitative analysis of the Ga-68-BZH3 kinetics can be used to predict the receptor expression of BB(2) in gliomas based on k1 of the compartment analysis. However, this study is limited to the expression data on the mRNA level and further studies are needed to assess the correlation of gene expression on the protein level.


Using human rights for sexual and reproductive health: improving legal and regulatory frameworks.
This paper describes the development of a tool that uses human rights concepts and methods to improve relevant laws, regulations and policies related to sexual and reproductive health. This tool aims to improve awareness and understanding of States' human rights obligations. It includes a method for systematically examining the status of vulnerable groups, involving non-health sectors, fostering a genuine process of civil society participation and developing recommendations to address regulatory and policy barriers to sexual and reproductive health with a clear assignment of responsibility. Strong leadership from the ministry of health, with support from the World Health Organization or other international partners, and the serious engagement of all involved in this process can strengthen the links between human rights and sexual and reproductive health, and contribute to national achievement of the highest attainable standard of health.


Electrochemical DNA biosensors based on long-range electron transfer: investigating the efficiency of a fluidic channel microelectrode compared to an ultramicroelectrode in a two-electrode setup.
Here, we describe the transposition of an ultramicroelectrode (UME) setup into a microfluidic chip configuration for DNA biosensors. The hydrodynamic properties of the fluidic channel microelectrode were screened with an [Fe(iii)(CN)6]3-/[Fe(ii)(CN)6]4- redox couple by cyclic voltammetry to provide a basis for further biological processes. A 23-base DNA probe was self-assembled into a monolayer on gold microelectrodes both in classical configuration and integrated in a microfluidic setup. Special interest was focused on the DNA target mimicking the liver-specific micro-ribonucleic acid 122 (miRNA122). Long-range electron transfer was chosen for transducing the hybridization. This direct transduction was indeed significantly enhanced after hybridization due to DNA-duplex π-stacking and the use of redox methylene blue as a DNA intercalator. Quantification of the target was deduced from the resulting electrical signal characterized by cyclic voltammetry. The limit of detection for DNA hybridization was 0.1 fM in stopped flow experiments, where it can reach 1 aM over a 0.5 μL s-1 flow rate, a value 104-fold lower than the one measured with a conventional UME dipped into an electrolyte droplet under the same analytical conditions. An explanation was that forced convection drives more biomolecules to the area of detection even if a balance between the speed of collection and the number of biomolecules collected has been found. The latter point is discussed here along with an attempt to explain why the sensor has reached such an unexpected value for the limit of detection.


Measuring giant anisotropy in paramagnetic transition metal complexes with relevance to single-ion magnetism.
"Giant magnetic anisotropy" is a phenomenon identified fairly recently in coordination complexes of nd- and nf-block ions. It can have different origins but is commonly (if not always correctly) described through zero-field splitting (ZFS), which can reach values of 100 cm-1 and beyond. This magnitude of anisotropy imparts on some of the complexes in question properties akin to single-ion magnets (SIMs). As a parameter of great importance, it needs to be accurately measured, a process that is far from being trivial. In this perspective article we give an overview and brief characterisation of the many experimental methods employed to that purpose, highlighting the strengths and weaknesses of each.


Hydrogen Embrittlement at Cleavage Planes and Grain Boundaries in Bcc Iron-Revisiting the First-Principles Cohesive Zone Model.
Hydrogen embrittlement, which severely affects structural materials such as steel, comprises several mechanisms at the atomic level. One of them is hydrogen enhanced decohesion (HEDE), the phenomenon of H accumulation between cleavage planes, where it reduces the interplanar cohesion. Grain boundaries are expected to play a significant role for HEDE, since they act as trapping sites for hydrogen. To elucidate this mechanism, we present the results of first-principles studies of the H effect on the cohesive strength of α-Fe single crystal (001) and (111) cleavage planes, as well as on the Σ5(310)[001] and Σ3(112)[11¯0] symmetrical tilt grain boundaries. The calculated results show that, within the studied range of concentrations, the single crystal cleavage planes are much more sensitive to a change in H concentration than the grain boundaries. Since there are two main types of procedures to perform ab initio tensile tests, different in whether or not to allow the relaxation of atomic positions, which can affect the quantitative and qualitative results, these methods are revisited to determine their effect on the predicted cohesive strength of segregated interfaces.


Advances in Campylobacter biology and implications for biotechnological applications.
Campylobacter jejuni is a major foodborne pathogen of animal origin and a leading cause of bacterial gastroenteritis in humans. During the past decade, especially since the publication of the first C. jejuni genome sequence, major advances have been made in understanding the pathobiology and physiology of this organism. It is apparent that C. jejuni utilizes sophisticated mechanisms for effective colonization of the intestinal tracts in various animal species. Although Campylobacter is fragile in the environment and requires fastidious growth conditions, it exhibits great flexibility in the adaptation to various habitats including the gastrointestinal tract. This high adaptability is attributable to its genetically, metabolically and phenotypically diverse population structure and its ability to change in response to various challenges. Unlike other enteric pathogens, such as Escherichia coli and Salmonella, Campylobacter is unable to utilize exogenous glucose and mainly depends on the catabolism of amino acids as a carbon source. Campylobacter proves highly mutable in response to antibiotic treatments and possesses eukaryote-like dual protein glycosylation systems, which modify flagella and other surface proteins with specific sugar structures. In this review we will summarize the distinct biological traits of Campylobacter and discuss the potential biotechnological approaches that can be developed to control this enteric pathogen.


Gnotobiotic rainbow trout (Oncorhynchus mykiss) model reveals endogenous bacteria that protect against Flavobacterium columnare infection.
The health and environmental risks associated with antibiotic use in aquaculture have promoted bacterial probiotics as an alternative approach to control fish infections in vulnerable larval and juvenile stages. However, evidence-based identification of probiotics is often hindered by the complexity of bacteria-host interactions and host variability in microbiologically uncontrolled conditions. While these difficulties can be partially resolved using gnotobiotic models harboring no or reduced microbiota, most host-microbe interaction studies are carried out in animal models with little relevance for fish farming. Here we studied host-microbiota-pathogen interactions in a germ-free and gnotobiotic model of rainbow trout (Oncorhynchus mykiss), one of the most widely cultured salmonids. We demonstrated that germ-free larvae raised in sterile conditions displayed no significant difference in growth after 35 days compared to conventionally-raised larvae, but were extremely sensitive to infection by Flavobacterium columnare, a common freshwater fish pathogen causing major economic losses worldwide. Furthermore, re-conventionalization with 11 culturable species from the conventional trout microbiota conferred resistance to F. columnare infection. Using mono-re-conventionalized germ-free trout, we identified that this protection is determined by a commensal Flavobacterium strain displaying antibacterial activity against F. columnare. Finally, we demonstrated that use of gnotobiotic trout is a suitable approach for the identification of both endogenous and exogenous probiotic bacterial strains protecting teleostean hosts against F. columnare. This study therefore establishes an ecologically-relevant gnotobiotic model for the study of host-pathogen interactions and colonization resistance in farmed fish.


Improved ex vivo method for microbiocidal activity across vertebrate species.
The field of ecoimmunology is currently undergoing rapid expansion, whereby biologists from a wide range of ecological disciplines are increasingly interested in assessing immunocompetence in their study organisms. One of the key challenges to researchers is determining what eco-immune measures to use in a given experiment. Moreover, there are limitations depending on study species, requirements for specific antibodies, and relevance of the methodology to the study organism. Here we introduce an improved ex vivo method for microbiocidal activity across vertebrate species. The utility of this assay is that it determines the ability of an organism to remove a pathogen that could be encountered in the wild, lending ecological relevancy to the technique. The applications of this microbiocidal assay are broad, as it is readily adaptable to different types of microbes as well as a wide variety of study species. We describe a method of microbiocidal analysis that will enable researchers across disciplines to effectively employ this method to accurately quantify microbial killing ability, using readily available microplate absorbance readers.


Diversity analysis of 80,000 wheat accessions reveals consequences and opportunities of selection footprints.
Undomesticated wild species, crop wild relatives, and landraces represent sources of variation for wheat improvement to address challenges from climate change and the growing human population. Here, we study 56,342 domesticated hexaploid, 18,946 domesticated tetraploid and 3,903 crop wild relatives in a massive-scale genotyping and diversity analysis. Using DArTseqTM technology, we identify more than 300,000 high-quality SNPs and SilicoDArT markers and align them to three reference maps: the IWGSC RefSeq v1.0 genome assembly, the durum wheat genome assembly (cv. Svevo), and the DArT genetic map. On average, 72% of the markers are uniquely placed on these maps and 50% are linked to genes. The analysis reveals landraces with unexplored diversity and genetic footprints defined by regions under selection. This provides fertile ground to develop wheat varieties of the future by exploring specific gene or chromosome regions and identifying germplasm conserving allelic diversity missing in current breeding programs.


LC-MS/MS method for the quantification of new psychoactive substances and evaluation of their urinary detection in humans for doping control analysis.
The constant legal adaptation of new psychoactive substances (NPS), challenges their evaluation in different fields. In sports, NPS are prohibited in competition with a reporting limit (RL) of 50 ng/mL for the parent compound or a metabolite. However, there is a lack of comprehensive methodologies and excretion studies for monitoring NPS. This work aims to develop an analytical methodology for the NPS quantification and to evaluate the suitability of monitoring the urinary parent stimulants after NPS misuse. A method for the quantification of 14 common NPS was developed and validated. The method was found to be linear in the range 1-1000 ng/mL, and was shown to be accurate and precise. A lowest limit of quantification (LLOQ) of 1 ng/mL was established for all analytes except for benzylpiperazine (5 ng/mL). The method was able to confirm the identity of the analytes at the LLOQ for most NPS. The methodology was applied to the quantification of the parent compound in urine samples collected from an observational study where several healthy volunteers (n ≥ 6 per drug) ingested active doses of mephedrone (MEPH), methylone (MDMC), 2,5-dimetoxy-4-ethylphenetylamine (2C-E), or 6-(2-aminopropyl)benzofuran (6-APB). It was observed that for MDMC and 6-APB, the quantification of the urinary parent drug at the current RL is a proper strategy for detecting their misuse. However, this strategy seems to be insufficient for evaluating MEPH and 2C-E misuse. Monitoring the most abundant metabolite of MEPH (4'-carboxy-MEPH) and the reduction of the RL to 10 ng/mL for the 2C-E evaluation are proposed.


Extreme size and sequence variation in the ITS rDNA of Bremia lactucae.
Bremia lactucae Regel (Chromista, Peronosporaceae) is an economically destructive pathogen, which causes downy mildew disease on lettuce (Lactuca sativa L.) worldwide. The ribosomal internal transcribed spacer (ITS) of Bremia lactucae isolates was analyzed for the first time. The ITS region of lettuce downy mildew was observed to have a size of 2458 bp; thereby, having one of the longest ITS sizes recorded to date. The majority of the extremely large sized ITS2 length of 2086 was attributed to the additional presences of nine repetitive elements with lengths of 179-194 bp, which between them shared the low homology of 48-69%. Comparison of the ITS2 sequences with the B. lactucae isolates from other host plants showed that isolates present on Lactuca sativa were distinct from those on L. indica var. laciniata, as well as Hemistepta and Youngia. We suggest the high degree of sequence heterogeneity exhibited in the ITS2 region of B. lactucae may warrant the specific detection and diagnosis of this destructive pathogen or its division into several distinct species.


Passive erbium-doped fiber seed photon generator for high-power Er(3+)-doped fiber fluorescent sources with an 80-nm bandwidth.
A novel structure that converts wasted backward amplified spontaneous emission (ASE) to seed photons for the amplifier stage is suggested for a high-power erbium-doped fiber (EDF) broadband source. A considerable increase in output power and bandwidth extension was achieved by placement of a segment of passive EDF in front of the amplifier stage, thus recycling backward ASE as the secondary pumping source for the passive EDF seed photon generator. Experimental results showed a dramatic increase in output ASE power of more than 10 dB for most radiation bands from 1540 to 1620 nm with the simple addition of an unpumped EDF segment to the ordinary fluorescence-source structure.


Validation of self-reported history of hysterectomy and oophorectomy among women in an integrated group practice setting.
Classification of menopause status often relies on self-report and is centrally important to research and clinical practice. This study was designed to assess the validity of self-reported hysterectomy and oophorectomy. A validation study of self-reported surgical menopause was conducted using survey data and electronic medical records from women enrolled in the Breast Cancer Screening Program within an integrated group practice in Washington State. Sensitivity of self-reported surgical history was estimated from questionnaire data among women with a history of hysterectomy (n = 1,935) and/or oophorectomy (n = 1,010) per medical records. Positive predictive values were quantified by reviewing medical records for a subset of women who self-reported a hysterectomy and/or oophorectomy (n = 122). Women self-reported hysterectomy history with great accuracy (sensitivity, 91%; positive predictive value, 97%) but were less accurate in reporting oophorectomy history (sensitivity of bilateral oophorectomy, 64%; positive predictive value, 100% and 73% for bilateral and unilateral oophorectomy, respectively). Among women self-reporting a unilateral oophorectomy, 19% had had both ovaries removed. Self-report is a valid data collection tool for hysterectomy history, but care should be taken in querying for and interpreting self-reported oophorectomy history for determining menopause status.


Making a decision to forgive.
Prominent models and interventions designed to promote forgiveness have distinguished one's decision to forgive from achieving forgiveness as an end state, but because of a lack of a strong measure, there is a weak research base on making a decision to forgive. Thus, in three studies, the authors developed the Decision to Forgive Scale (DTFS) and examined evidence for its reliability and construct validity. The article focused on distinguishing making a decision to forgive from achieved level of forgiveness. Scores on the DTFS showed evidence of reliability, with Cronbach's alpha coefficients ranging from .92 to .94, and a 1-week temporal stability coefficient of .68. Using several strategies, the authors demonstrated that the DTFS is empirically distinct from the Transgression-Related Interpersonal Motivations scale (TRIM; McCullough et al., 1998). Namely, a 3-factor confirmatory factor analysis that included the DTFS and the 2 TRIM subscales showed excellent fit, suggesting these instruments assess 3 different constructs. The DTFS was only moderately related to the TRIM subscales, was more strongly related to stage of change than the TRIM, and predicted subsequent TRIM scores in a cross-lagged model. Finally, although decisions to forgive generally suggested greater forgiveness, these constructs interacted to predict existential distress. Namely, as decisional forgiveness increased, revenge was more strongly related to existential distress. Overall, the DTFS shows considerable promise for further clinical and basic research applications.


Reading level attenuates differences in neuropsychological test performance between African American and White elders.
The current study sought to determine if discrepancies in quality of education could explain differences in cognitive test scores between African American and White elders matched on years of education. A comprehensive neuropsychological battery was administered to a sample of African American and non-Hispanic White participants in an epidemiological study of normal aging and dementia in the Northern Manhattan community. All participants were diagnosed as nondemented by a neurologist, and had no history of Parkinson's disease, stroke, mental illness, or head injury. The Reading Recognition subtest from the Wide Range Achievement Test-Version 3 was used as an estimate of quality of education. A MANOVA revealed that African American elders obtained significantly lower scores than Whites on measures of word list learning and memory, figure memory, abstract reasoning, fluency, and visuospatial skill even though the groups were matched on years of education. However, after adjusting the scores for WRAT-3 reading score, the overall effect of race was greatly reduced and racial differences on all tests (except category fluency and a drawing measure) became nonsignificant. These findings suggest that years of education is an inadequate measure of the educational experience among multicultural elders, and that adjusting for quality of education may improve the specificity of certain neuropsychological measures.


Optimization of duplex stability and terminal asymmetry for shRNA design.
Prediction of efficient oligonucleotides for RNA interference presents a serious challenge, especially for the development of genome-wide RNAi libraries which encounter difficulties and limitations due to ambiguities in the results and the requirement for significant computational resources. Here we present a fast and practical algorithm for shRNA design based on the thermodynamic parameters. In order to identify shRNA and siRNA features universally associated with high silencing efficiency, we analyzed structure-activity relationships in thousands of individual RNAi experiments from publicly available databases (ftp://ftp.ncbi.nlm.nih.gov/pub/shabalin/siRNA/si_shRNA_selector/). Using this statistical analysis, we found free energy ranges for the terminal duplex asymmetry and for fully paired duplex stability, such that shRNAs or siRNAs falling in both ranges have a high probability of being efficient. When combined, these two parameters yield a approximately 72% success rate on shRNAs from the siRecords database, with the target RNA levels reduced to below 20% of the control. Two other parameters correlate well with silencing efficiency: the stability of target RNA and the antisense strand secondary structure. Both parameters also correlate with the short RNA duplex stability; as a consequence, adding these parameters to our prediction scheme did not substantially improve classification accuracy. To test the validity of our predictions, we designed 83 shRNAs with optimal terminal asymmetry, and experimentally verified that small shifts in duplex stability strongly affected silencing efficiency. We showed that shRNAs with short fully paired stems could be successfully selected by optimizing only two parameters: terminal duplex asymmetry and duplex stability of the hypothetical cleavage product, which also relates to the specificity of mRNA target recognition. Our approach performs at the level of the best currently utilized algorithms that take into account prediction of the secondary structure of the target and antisense RNAs, but at significantly lower computational costs. Based on this study, we created the si-shRNA Selector program that predicts both highly efficient shRNAs and functional siRNAs (ftp://ftp.ncbi.nlm.nih.gov/pub/shabalin/siRNA/si_shRNA_selector/).


Potential of Plant-sourced Phenols for Inflammatory Bowel Disease.
Inflammatory bowel disease (IBD) is an uncontrolled chronic inflammatory intestinal disorder, which requires medications for long-term therapy. Facing the challenges of severe side effects and drug resistance of conventional medications, to develop the strategies meet the stringent safety and effectiveness in the long-term treatment are urgent in the clinics. In this regard, a growing body of evidence confirms plant-sourced phenols, such as flavonoids, catechins, stilbenes, coumarins, quinones, lignans, phenylethanoids, cannabinoid phenols, tannins, phenolic acids and hydroxyphenols, exert potent protective benefits with fewer undesirable effects in conditions of acute or chronic intestinal inflammation through improvement of colonic oxidative and pro-inflammatory status, preservation of the epithelial barrier function and modulation of gut microbiota. In this review, the great potential of plant-sourced phenols and their action mechanisms for the treatment or prevention of IBD in recent research are summarized, which may help further development of new preventive/adjuvant regimens for IBD.


Cardiac perfusion imaging using hyperpolarized (13)C urea using flow sensitizing gradients.
To demonstrate the feasibility of imaging the first passage of a bolus of hyperpolarized (13)C urea through the rodent heart using flow-sensitizing gradients to reduce signal from the blood pool. A flow-sensitizing bipolar gradient was optimized to reduce the bright signal within the cardiac chambers, enabling improved contrast of the agent within the tissue capillary bed. The gradient was incorporated into a dynamic golden angle spiral (13)C imaging sequence. Healthy rats were scanned during rest (n = 3) and under adenosine stress-induced hyperemia (n = 3). A two-fold increase in myocardial perfusion relative to rest was detected during adenosine stress-induced hyperemia, consistent with a myocardial perfusion reserve of two in rodents. The new pulse sequence was used to obtain dynamic images of the first passage of hyperpolarized (13)C urea in the rodent heart, without contamination from bright signal within the neighboring cardiac lumen. This probe of myocardial perfusion is expected to enable new hyperpolarized (13)C studies in which the cardiac metabolism/perfusion mismatch can be identified. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance.


Approximating covering problems by randomized search heuristics using multi-objective models.
The main aim of randomized search heuristics is to produce good approximations of optimal solutions within a small amount of time. In contrast to numerous experimental results, there are only a few theoretical explorations on this subject. We consider the approximation ability of randomized search heuristics for the class of covering problems and compare single-objective and multi-objective models for such problems. For the VertexCover problem, we point out situations where the multi-objective model leads to a fast construction of optimal solutions while in the single-objective case, no good approximation can be achieved within the expected polynomial time. Examining the more general SetCover problem, we show that optimal solutions can be approximated within a logarithmic factor of the size of the ground set, using the multi-objective approach, while the approximation quality obtainable by the single-objective approach in expected polynomial time may be arbitrarily bad.


Review article: Emergency department data sharing to reduce alcohol-related violence: a systematic review of the feasibility and effectiveness of community-level interventions.
The present paper aims to review current evidence for the effectiveness and/or feasibility of using inter-agency data sharing of ED recorded assault information to direct interventions reducing alcohol-related or nightlife assaults, injury or violence. Potential data-sharing partners involve police, local council, liquor licensing regulators and venue management. A systematic review of the peer-reviewed literature was conducted. The initial search discovered 19,506 articles. After removal of duplicates and articles not meeting review criteria, n = 8 articles were included in quantitative and narrative synthesis. Seven of eight studies were conducted in UK EDs, with the remaining study presenting Australian data. All studies included in the review deemed data sharing a worthwhile pursuit. All studies attempting to measure intervention effectiveness reported substantial reductions of assaults and ED attendances post-intervention, with one reporting no change. Negative logistic feasibility concerns were minimal, with general consensus among authors being that data-sharing protocols and partnerships could be easily implemented into modern ED triage systems, with minimal cost, staff workload burden, impact to patient safety, service and anonymity, or risk of harm displacement to other licensed venues, or increase to length of patient stay. However, one study reported a potential harm displacement effect to streets surrounding intervention venues. In future, data-sharing systems should triangulate ED, police and ambulance data sources, and assess intervention effectiveness using randomised controlled trials that account for variations in venue capacity, fluctuations in ED attendance and population levels, seasonal variations in assault and injury, and control for concurrent interventions.


From supramolecular polymers to multi-component biomaterials.
The most striking and general property of the biological fibrous architectures in the extracellular matrix (ECM) is the strong and directional interaction between biologically active protein subunits. These fibers display rich dynamic behavior without losing their architectural integrity. The complexity of the ECM taking care of many essential properties has inspired synthetic chemists to mimic these properties in artificial one-dimensional fibrous structures with the aim to arrive at multi-component biomaterials. Due to the dynamic character required for interaction with natural tissue, supramolecular biomaterials are promising candidates for regenerative medicine. Depending on the application area, and thereby the design criteria of these multi-component fibrous biomaterials, they are used as elastomeric materials or hydrogel systems. Elastomeric materials are designed to have load bearing properties whereas hydrogels are proposed to support in vitro cell culture. Although the chemical structures and systems designed and studied today are rather simple compared to the complexity of the ECM, the first examples of these functional supramolecular biomaterials reaching the clinic have been reported. The basic concept of many of these supramolecular biomaterials is based on their ability to adapt to cell behavior as a result of dynamic non-covalent interactions. In this review, we show the translation of one-dimensional supramolecular polymers into multi-component functional biomaterials for regenerative medicine applications.


Endocrine-disrupting chemicals and obesity development in humans: a review.
This study reviewed the literature on the relations between exposure to chemicals with endocrine-disrupting abilities and obesity in humans. The studies generally indicated that exposure to some of the endocrine-disrupting chemicals was associated with an increase in body size in humans. The results depended on the type of chemical, exposure level, timing of exposure and gender. Nearly all the studies investigating dichlorodiphenyldichloroethylene (DDE) found that exposure was associated with an increase in body size, whereas the results of the studies investigating polychlorinated biphenyl (PCB) exposure were depending on dose, timing and gender. Hexachlorobenzene, polybrominated biphenyls, beta-hexachlorocyclohexane, oxychlordane and phthalates were likewise generally associated with an increase in body size. Studies investigating polychlorinated dibenzodioxins and polychlorinated dibenzofurans found either associations with weight gain or an increase in waist circumference, or no association. The one study investigating relations with bisphenol A found no association. Studies investigating prenatal exposure indicated that exposure in utero may cause permanent physiological changes predisposing to later weight gain. The study findings suggest that some endocrine disruptors may play a role for the development of the obesity epidemic, in addition to the more commonly perceived putative contributors.


Immortalization and altered differentiation of human keratinocytes in vitro by the E6 and E7 open reading frames of human papillomavirus type 18.
The E6-E7 region of human papillomavirus types 16 and 18 is selectively retained and expressed in cervical carcinoma cells. In cultured human keratinocytes, expression of the E6 and E7 open reading frames of human papillomavirus type 18, under the control of its homologous promoter, resulted in high-frequency immortalization. Furthermore, by using a system that allows for stratification of keratinocytes in vitro (raft system), we observed that the morphological differentiation of these E6-E7 immortalized cells was altered such that parabasal cells extended throughout most of the epithelium, with abnormal nuclei present in the upper regions. Examination of E6-E7-expressing cell lines in the raft system at a later passage revealed that complete loss of morphological differentiation had occurred. E7 alone was a much less effective immortalizing agent than E6 and E7 together and acted only minimally to alter morphological differentiation in vitro. No such activities were found for E6 alone. High-frequency transformation of human epithelial cells thus appears to require expression of both E6 and E7 gene products.


Endoscopic management of sinonasal tumors: a preliminary report.
The increased experience with the endoscopic approach to sinonasal inflammatory disease has resulted in the increased use of endoscopes to manage many different sinonasal pathologies. A chart review of patients with sinonasal tumors treated with primary endoscopic management, from January, 1993 to November, 2003 was performed. Forty patients were identified (26 men and 14 women). The mean age was 53.2 years, and the mean follow-up was 31.1 months. For benign tumors, 24 patients were identified with a mean age of 50.7 years, a mean follow-up of 17.5 months, and a recurrence rate of 4.2%. For malignant tumors, 16 patients were identified, with a mean age of 57.3 years, a mean follow-up of 51.5 months, and a recurrence rate of 31.3%. The overall survival rate was 87.5%. Endoscopic surgical excision of selective sinonasal tumors may be an effective therapeutic modality. In some cases, adjuvant external procedures may be required based on tumor location.


Cognitive milieu therapy and physical activity: experiences of mastery and learning among patients with dual diagnosis.
During the last decade, there has been a growing interest in implementing cognitive milieu therapy (CMT) in psychiatric institutions. However, there is a lack of systematic evaluations from patients' point of view. The aim of this study was to explore and describe patient perceptions of essential experiences of mastery, learning alternative ways of thinking, and acquiring new skills through CMT and physical activity in an inpatient setting. Qualitative interviews were carried out with 20 patients with dual diagnosis. A hermeneutic - phenomenological approach was used in the data collection and analysis. The results showed that the learning climate in the unit was important. This included a proactive attitude from the staff, focusing on cooperation on equal terms between patients and staff, and a professional methodological approach through CMT. The optimal balance between staff-induced activities and patient initiatives was not easy to obtain. Patients appreciated both the education provided by the staff and learning from other patients. The cognitive method was sometimes experienced as too theoretical and difficult to understand. Physical activity, however, was experienced as 'concrete' and providing practical knowledge. It motivated patients to establish new habits and provided opportunities for the development of mastery together with other patients.


A Pt-cluster-based heterogeneous catalyst for homogeneous catalytic reactions: X-ray absorption spectroscopy and reaction kinetic studies of their activity and stability against leaching.
The design and development of metal-cluster-based heterogeneous catalysts with high activity, selectivity, and stability under solution-phase reaction conditions will enable their applications as recyclable catalysts in large-scale fine chemicals production. To achieve these required catalytic properties, a heterogeneous catalyst must contain specific catalytically active species in high concentration, and the active species must be stabilized on a solid catalyst support under solution-phase reaction conditions. These requirements pose a great challenge for catalysis research to design metal-cluster-based catalysts for solution-phase catalytic processes. Here, we focus on a silica-supported, polymer-encapsulated Pt catalyst for an electrophilic hydroalkoxylation reaction in toluene, which exhibits superior selectivity and stability against leaching under mild reaction conditions. We unveil the key factors leading to the observed superior catalytic performance by combining X-ray absorption spectroscopy (XAS) and reaction kinetic studies. On the basis of the mechanistic understandings obtained in this work, we also provide useful guidelines for designing metal-cluster-based catalyst for a broader range of reactions in the solution phase.


How often is the diagnosis of the permanent vegetative state incorrect? A review of the evidence.
Some research suggests that 40% of people in the vegetative state are misdiagnosed. This review investigates the frequency, nature and causes of reported misdiagnosis of patients in the vegetative state, focusing on the nature of the error. It is a systematic review of all relevant literature, using references from key papers identified. The data are summarized in tables. Five clinical studies of the rate of misdiagnosis in practice were identified, encompassing 236 patients in the vegetative state of whom 80 (34%) were reclassified as having some awareness, often minimal. The studies often included patients in the recovery phase after acute injury, and were poorly reported. Five systematic reviews of signs and technologically based neurophysiological tests were identified, and they showed that most studies were small, lacked accurate or important details, and were subject to bias. Studies were not replicated. Many signs and tests did not differ between people in the vegetative state and in the minimally conscious state, and those that did were unable to diagnose an individual patient. The few single case reports suggest that failure to ensure an accurate diagnosis of the underlying neurological damage and dysfunction could, rarely, lead to significant misdiagnosis usually in patients who had brain-stem damage with little thalamic or cortical damage. Significant misdiagnosis of awareness, with an apparently 'vegetative' patient having good awareness, is rare. Careful neurological assessment of the cause and routine measurement of awareness using the Coma Recovery Scale - Revised should further reduce mistakes.


Treatment of arm, neck, and/or shoulder complaints in physical therapy practice.
This is a prospective cohort study with a follow-up period of 6 months. To describe the interventions applied by physical therapists in treating patients with complaints of the arm, neck, and/or shoulder. Complaints of the arm, neck, and/or shoulder occur frequently and are usually treated within primary care. Many patients with these complaints are treated with physical therapy. During a 1-year period, the participating physiotherapists included new consulters with musculoskeletal complaints of the upper extremity. The patients completed a questionnaire at baseline, and the physical therapists completed a treatment record for up to 6 months. Of the 624 patients included in the cohort, physical therapists provided treatment data of 619 patients. The main treatment consisted of exercise therapy (93%) and massage (87%), or a combination of both. More patients with specific complaints received physical methods, and patients with nonspecific complaints were treated with manipulation techniques. The primary treatment of complaints the of arm, neck, and/or shoulder in physiotherapy practice consists of exercise therapy and massage therapy, mostly being a combination of both. Future studies should focus on the effectiveness of exercise therapies, combined or not combined with massage, for patients with complaints of the arm, neck, and/or shoulder.


PERK activation at low glucose concentration is mediated by SERCA pump inhibition and confers preemptive cytoprotection to pancreatic β-cells.
Protein kinase R-like ER kinase (PERK) is activated at physiologically low glucose concentrations in pancreatic β-cells. However, the molecular mechanisms by which PERK is activated under these conditions and its role in β-cell function are poorly understood. In this report, we investigated, in dispersed rat islets of Langerhans and mouse insulinoma-6 (MIN6) cells, the relationship between extracellular glucose concentration, the free endoplasmic reticulum (ER) calcium concentration ([Ca(2+)](ER)) measured directly using an ER targeted fluorescence resonance energy transfer-based calcium sensor, and the activation of PERK. We found that a decrease in glucose concentration leads to a concentration-dependent reduction in [Ca(2+)](ER) that parallels the activation of PERK and the phosphorylation of its substrate eukaryotic initiation factor-2α. We provide evidence that this decrease in [Ca(2+)](ER) is caused by a decrease in sarcoplasmic/ER Ca(2+)-ATPase pump activity mediated by a reduction in the energy status of the cell. Importantly, we also report that PERK-dependent eukaryotic initiation factor-2α phosphorylation at low glucose concentration plays a significant role in 1) the regulation of both proinsulin and global protein synthesis, 2) cell viability, and 3) conferring preemptive cytoprotection against ER stress. Taken together, these results provide evidence that a decrease in the ATP/energy status of the cell in response to a decrease in glucose concentration results in sarcoplasmic/ER Ca(2+)-ATPase pump inhibition, the efflux of Ca(2+) from the ER, and the activation of PERK, which plays an important role in both pancreatic β-cell function and survival.


Antimicrobial susceptibility of stenotrophomonas maltophilia isolates from Korea, and the activity of antimicrobial combinations against the isolates.
The aim of this study was to determine antimicrobial susceptibility of recent clinical Stenotrophomonas maltophilia isolates from Korea, and to compare the activity levels of several combinations of antimicrobials. A total of 206 non-duplicate clinical isolates of S. maltophilia was collected in 2010 from 11 university hospitals. Antimicrobial susceptibility testing was performed using the Clinical Laboratory Standards Institute agar dilution method. In vitro activity of antimicrobial combinations was tested using the checkerboard method. The susceptibility rates to trimethoprim-sulfamethoxazole and minocycline were 96% and 99%, respectively. The susceptibility rate to levofloxacin was 64%. All of four antimicrobial combinations showed synergy against many S. maltophilia isolates. A combination of trimethoprim-sulfamethoxazole plus ticarcillin-clavulanate was most synergistic among the combinations. None of the combinations showed antagonistic activity. Therefore, some of the combinations may be more useful than individual drugs in the treatment of S. maltophilia infection. Further clinical studies are warranted to validate our in vitro test results.


Transient coexpression of individual genes encoded by the triple gene block of potato mop-top virus reveals requirements for TGBp1 trafficking.
TGBp1, TGBp2, and TGBp3, three plant virus movement proteins encoded by the "triple gene block" (TGB), may act in concert to facilitate cell-to-cell transport of viral RNA genomes. Transient expression of Potato mop-top virus (genus Pomovirus) movement proteins was used as a model to reconstruct interactions between TGB proteins. In bombarded epidermal cells of Nicotiana benthamiana, green fluorescent protein (GFP)-TGBp1 was distributed uniformly. However, in the presence of TGBp2 and TGBp3, GFP-TGBp1 was directed to intermediate bodies at the cell periphery, and to cell wall-embedded punctate bodies. Moreover, GFP-TGBp1 migrated into cells immediately adjacent to the bombarded cell. These data suggest that TGBp2 and TGBp3 mediate transport of GFP-TGBp1 to and through plasmodesmata. Mutagenesis of TGBp1 suggested that the NTPase and helicase activities of TGBp1 were not required for its transport to intermediate bodies directed by TGBp2 and TGBp3, but these activities were essential for the protein association with cell wall-embedded punctate bodies and translocation of TGBpl to neighboring cells. The C-terminal region of TGBp1 was critical for trafficking mediated by TGBp2 and TGBp3. Mutation analysis also suggested an involvement of the TGBp2 C-terminal region in interactions with TGBp1.


Quantitative 3D micro-CT imaging of human lung tissue.
To assess the feasibility of micro-CT for obtaining quantitative volumetric and morphologic information of changes in soft tissue, respiratory tracts and vascularization in fibrotic, emphysematous and non-diseased human lung specimens. Specimens from autopsy or lung explantation with lung fibrosis of UIP pattern (n = 22) or centrilobular emphysema (n = 10) were scanned by micro-CT and compared to controls (n = 22). Imaging was performed subsequent to intravascular contrast enhancement for the assessment of the vascular volume fraction. The soft tissue and air fraction were quantified after the fixation of ventilated lungs followed by tissue contrast enhancement using osmium. Aiming an artifact-free 3 D reconstruction of lung acini, synchrotron-based micro-CT scans of specimens with emphysema (n = 5) and non-diseased tissue (n = 6) was performed. Micro-CT imaging was complemented by histology for the demonstration of comparable findings. Quantitative analysis showed a significant increase of the soft tissue fraction, equivalent to a decrease of the air fraction in fibrotic lungs compared to controls (p < 0.001) and a significant reduction of the vascular volume fraction compared to controls (p < 0.02). Specimens with emphysema demonstrated a significant increase of the air fraction with a decrease in soft tissue compared to controls (p < 0.001). 3 D reconstructions of lung acini worked successfully in non-diseased tissue but failed in fibrotic and emphysematous lungs. Our findings indicate micro-CT's technical feasibility to assess quantitative and morphological data from diseased and non-diseased human lung specimens.


Angiotensin II receptor type 1 A1166C modifies the association between angiotensinogen M235T and chronic kidney disease.
Single nucleotide polymorphisms (SNPs) in renin-angiotensin system (RAS) genes are associated with RAS imbalance and chronic kidney disease (CKD). We performed a case-control study and meta-analysis to investigate the association between angiotensinogen (AGT) M235T polymorphism and CKD. A total of 634 patients with end-stage renal disease and 739 healthy controls were studied. We also searched PubMed and the Cochrane Library to identify prospective observational studies published before December 2015. We found that the TT and MT genotypes were associated with a higher risk of CKD than the MM genotype (odds ratio [OR]: 3.56; 95% confidence interval [CI]: 1.14-11.16 and OR: 2.93; 95% CI: 0.91-9.46, respectively). Thirty-eight study populations were included in the meta-analysis. The T allele was associated with a higher risk of CKD than the M allele in all populations (OR: 1.19; 95% CI: 1.08-1.32). The OR was 1.33 in Asians (95% CI: 1.06-1.67) and 1.10 in Caucasians (95% CI: 1.02-1.18). Evaluation of gene-gene and gene-environment interactions using epistasis analysis revealed an interaction between AGT M235T and angiotensin II receptor type 1 A1166C in CKD (OR: 0.767; 95% CI: 0.609-0.965). Genetic testing for CKD in high-risk individuals may be an effective strategy for CKD prevention.


An organotypic slice model for ex vivo study of neural, immune, and microbial interactions of mouse intestine.
Organotypic tissue slices provide seminatural, three-dimensional microenvironments for use in ex vivo study of specific organs and have advanced investigative capabilities compared with isolated cell cultures. Several characteristics of the gastrointestinal tract have made in vitro models for studying the intestine challenging, such as maintaining the intricate structure of microvilli, the intrinsic enteric nervous system, Peyer's patches, the microbiome, and the active contraction of gut muscles. In the present study, an organotypic intestinal slice model was developed that allows for functional investigation across regions of the intestine. Intestinal tissue slices were maintained ex vivo for several days in a physiologically relevant environment that preserved normal enterocyte structure, intact and proliferating crypt cells, submucosal organization, and muscle wall composure. Cell death was measured by a membrane-impermeable DNA binding indicator, ethidium homodimer, and less than 5% of cells were labeled in all regions of the villi and crypt epithelia at 24 h ex vivo. This tissue slice model demonstrated intact myenteric and submucosal neuronal plexuses and functional interstitial cells of Cajal to the extent that nonstimulated, segmental contractions occurred for up to 48 h ex vivo. To detect changes in physiological responses, slices were also assessed for segmental contractions in the presence and absence of antibiotic treatment, which resulted in slices with lesser or greater amounts of commensal bacteria, respectively. Segmental contractions were significantly greater in slices without antibiotics and increased native microbiota. This model renders mechanisms of neuroimmune-microbiome interactions in a complex gut environment available to direct observation and controlled perturbation.


Reconfigurable Coplanar Waveguide (CPW) and Half-Mode Substrate Integrated Waveguide (HMSIW) Band-Stop Filters Using a Varactor-Loaded Metamaterial-Inspired Open Resonator.
An open ring resonator (ORR) loaded with a varactor diode is designed and implemented in order to achieve high-performance tunable band-stop filters in planar technology with a compact size. This varactor-loaded ORR (VLORR) is versatile. It allows a shunt connection with different planar waveguide sections. In this paper, it has been connected to a coplanar waveguide (CPW) and a half-mode substrate integrated waveguide (HMSIW). As a reverse bias voltage is applied to the VLORR, a continuous tuning over the resulting stop-band can be achieved. To illustrate the possibilities of the VLORR, three prototypes have been designed, fabricated, and characterized. The three prototypes show an outstanding performance, with a rejection level at the resonant frequency and a tuning range greater than 12 dB and 85%, respectively. This VLORR has high potential value in microwave communication systems to eliminate unwanted signals.


Inhibition of metabotropic glutamate receptor signaling by the huntingtin-binding protein optineurin.
Huntington disease is caused by a polyglutamine expansion in the huntingtin protein (Htt) and is associated with excitotoxic death of striatal neurons. Group I metabotropic glutamate receptors (mGluRs) that are coupled to inositol 1,4,5-triphosphate formation and the release of intracellular Ca(2+) stores play an important role in regulating neuronal function. We show here that mGluRs interact with the Htt-binding protein optineurin that is also linked to normal pressure open angled glaucoma and, when expressed in HEK 293 cells, optineurin functions to antagonize agonist-stimulated mGluR1a signaling. We find that Htt is co-precipitated with mGluR1a and that mutant Htt functions to facilitate optineurin-mediated attenuation of mGluR1a signaling. In striatal cell lines derived from Htt(Q111/Q111) mutant knock-in mice mGluR5-stimulated inositol phosphate formation is also severely impaired when compared with striatal cells derived from Htt(Q7/Q7) knock-in mice. In addition, we show that a missense single nucleotide polymorphism optineurin H486R variant previously identified to be associated with glaucoma is selectively impaired in mutant Htt binding. Although optineurin H486R retains the capacity to bind to mGluR1a, optineurin H486R-dependent attenuation of mGluR1a signaling is not enhanced by the expression of mutant Htt. Because G protein-coupled receptor kinase 2 (GRK2) protein expression is relatively low in striatal tissue, we propose that optineurin may substitute for GRK2 in the striatum to mediate mGluR desensitization. Taken together, these studies identify a novel mechanism for mGluR desensitization and an additional biochemical link between altered glutamate receptor signaling and Huntington disease.


Description of a new Peronia species (Gastropoda: Eupulmonata: Onchidiidae) from Iran, Persian Gulf.
Peronia J. Fleming, 1822 is an eupulmonate slug genus with a wide distribution in the Indo-Pacific Ocean. Currently, nine species are considered as valid. However, molecular data indicate cryptic speciation and more species involved. Here, we present results on a new species found in the Persian Gulf, a subtropical region with harsh conditions such as elevated salinity and high temperature compared to the Indian Ocean. Peronia persiae sp. nov. is described based on molecular, histological, anatomical, micro-computer tomography and scanning electron microscopy data. ABGD, GMYC and bPTP analyses based on 16S rDNA and cytochrome oxidase I (COI) sequences of Peronia confirm the delimitation of the new species. Moreover, our 14 specimens were carefully compared with available information of other described Peronia species. Peronia persiae sp. nov. is distinct in a combination of characters, including differences in the genital (ampulla, prostate, penial hooks, penial needle) and digestive systems (lack of pharyngeal wall teeth, tooth shape in radula, intestine of type II).


Grain Scattering Noise Modeling and Its Use in the Detection and Characterization of Defects Using Ultrasonic Arrays.
In the field of ultrasonic array imaging for non-destructive testing (NDT), material structural noise caused by grain scattering is one of the main sources of error when characterizing defects that are found in the polycrystalline materials. The existence of grains can also severely affect the detection performance of ultrasonic testing, making small defects indistinguishable from the grain indications due to ultrasonic attenuation and backscatter. This paper proposes a model in which the statistical distribution of the defect data is obtained from different realizations of the grain structure. This statistical distribution, termed the defect+grains model in this paper, is shown to contain information that is needed for detection and characterization of defects. Hence, given a specific measurement configuration, the characterization result can be obtained by constructing a defect+grains model based on the multiple realizations of each possible defect and calculating their probability. The detection, classification, and sizing accuracy are shown to be predictable by quantifying the probabilities that an experimentally measured defect matches the different defect+grains models. This defect+grains modeling approach gives insight into the detection/characterization problem, leading to an evaluation of the fundamental limits of the achievable inspection performance.


Does implied community size predict likeability of a similar stranger?
Homophily, the tendency for people to cluster with similar others, has primarily been studied in terms of proximal, psychological causes, such as a tendency to have positive associations with people who share traits with us. Here we investigate whether homophily could be correlated with perceived group membership, given that sharing traits with other people might signify membership of a specific community. In order to investigate this, we tested whether the amount of homophily that occurs between strangers is dependent on the number of people they believe share the common trait (i.e. the size of group that the trait identifies). In two experiments, we show that more exclusive (smaller) groups evoke more positive ratings of the likeability of a stranger. When groups appear to be too inclusive (i.e. large) homophily no longer occurs, suggesting that it is not only positive associations with a trait that cause homophily, but a sense of the exclusiveness of a group is also important. These results suggest that group membership based on a variety of traits can encourage cohesion between people from diverse backgrounds, and may be a useful tool in overcoming differences between groups.


Definition and properties of disequilibria within nuclear-mitochondrial-chloroplast and other nuclear-dicytoplasmic systems.
We define and determine the interrelationships among five sets of disequilibrium parameters that measure two- and three-locus nonrandom associations in nuclear-dicytoplasmic systems. These assume a diploid nuclear locus and two haploid cytoplasmic loci, with special reference to nuclear-mitochondrial-chloroplast systems. Three sets of two-locus disequilibria measure the association between haplotypes at the two cytoplasmic loci (DMC) and associations between each cytoplasmic locus and nuclear alleles or genotypes (DM, D1M, D2M, D3M; DC, D1C, D2C, D3C). In addition, we present two classes of higher-order disequilibria that measure nonrandom allelic or genotypic associations involving all three loci. The first class quantifies associations between the nuclear locus and the two cytoplasmic loci taken jointly (DA/MC, DAA/MC, DAa/MC, Daa/MC, etc.), whereas the second measures only those associations remaining after all two-locus associations have been taken into account (DA/M/C, DAA/M/C, DAa/M/C, Daa/M/C). Based on combinations of these five sets of measures, we suggest a variety of parameterizations of three-locus, nuclear-dicytoplasmic systems. The dynamics of these disequilibria are then investigated under models of random and mixed mating, either with both cytoplasmic genomes inherited through the same parent or through opposite parents. Except for associations between the cytoplasmic haplotypes, which are constant when the two cytoplasmic genomes are inherited through the same parent, all disequilibria ultimately decay to zero. These randomizations do not necessarily occur monotonically, however, and in some cases are preceded by an initial increase in magnitude or sign change. For both inheritance patterns, the asymptotic decay rates are steadily retarded by increasing levels of self-fertilization. This behavior contrasts with that in the extreme case of complete selfing, for which only the heterozygote disequilibria always decay to zero. For all models considered, the dynamics of the two-locus cytonuclear subsystems are solely a function of the mating system, whereas the dynamical behavior and sign patterns of the cytoplasmic and three-locus disequilibria also depend strongly on the mode of cytoplasmic inheritance.


Insertion/deletion polymorphism in the angiotensin-I-converting enzyme gene is associated with coronary heart disease in IDDM patients with diabetic nephropathy.
Insulin-dependent diabetic (IDDM) patients with diabetic nephropathy have a highly increased morbidity and mortality from coronary heart disease. An insertion (I) /deletion (D) polymorphism in the angiotensin-I-converting enzyme (ACE) gene has been shown to be associated with coronary heart disease. Therefore, we have investigated the role of this ACE/ID polymorphism in 198 IDDM patients with diabetic nephropathy and 190 normoalbuminuric IDDM patients. The prevalence of myocardial infarction and other coronary heart disease was significantly elevated in patients with nephropathy, 19% (38/198) vs 8% (15/190), p < 0.001. In the nephropathic group 12 of 63 (19%), 23 of 95 (24%), and 3 of 40 (7.5%) patients with the DD, ID and II genotypes, respectively had a history of coronary heart disease, II vs DD and ID, p < 0.05 when compared to nephropathic patients without coronary heart disease. Multiple logistic regression analysis of the risk factors associated with coronary heart disease in univariate analysis revealed that the II genotype acts as an independent protective factor against coronary heart disease, odds ratio II/DD + ID 0.27 (95% confidence interval 0.07-0.97, p < 0.05). There was no difference in genotype or allele frequency (D/I) between patients with and without nephropathy, 0.56/0.44 in both groups, but plasma ACE concentration was elevated in patients with nephropathy 609 (151-1504) ng/ml as compared to patients with normoalbuminuria, 428 (55-1630) ng/ml, p < 0.001. We suggest that ACE/ID polymorphism may influence the frequency of life-threatening cardiac complications in IDDM patients suffering from diabetic nephropathy, a condition characterized by increased plasma ACE concentration.


Cardioacceleratory function of the neurohormone CCAP in the mosquito Anopheles gambiae.
Crustacean cardioactive peptide (CCAP) is a highly conserved arthropod neurohormone that is involved in ecdysis, hormone release and the modulation of muscle contractions. Here, we determined the CCAP gene structure in the malaria mosquito Anopheles gambiae, assessed the developmental expression of CCAP and its receptor and determined the role that CCAP plays in regulating mosquito cardiac function. RACE sequencing revealed that the A. gambiae CCAP gene encodes a neuropeptide that shares 100% amino acid identity with all sequenced CCAP peptides, with the exception of Daphnia pulex. Quantitative RT-PCR showed that expression of CCAP and the CCAP receptor displays a bimodal distribution, with peak mRNA levels in second instar larvae and pupae. Injection of CCAP revealed that augmenting hemocoelic CCAP levels in adult mosquitoes increases the anterograde and retrograde heart contraction rates by up to 28%, and increases intracardiac hemolymph flow velocities by up to 33%. Partial CCAP knockdown by RNAi had the opposite effect, decreasing the mosquito heart rate by 6%. Quantitative RT-PCR experiments showed that CCAP mRNA is enriched in the head region, and immunohistochemical experiments in newly eclosed mosquitoes detected CCAP in abdominal neurons and projections, some of which innervated the heart, but failed to detect CCAP in the abdomens of older mosquitoes. Instead, in older mosquitoes CCAP was detected in the pars lateralis, the subesophageal ganglion and the corpora cardiaca. In conclusion, CCAP has a potent effect on mosquito circulatory physiology, and thus heart physiology in this dipteran insect is under partial neuronal control.


Photothermal tomography for the functional and structural evaluation, and early mineral loss monitoring in bones.
Salient features of a new non-ionizing bone diagnostics technique, truncated-correlation photothermal coherence tomography (TC-PCT), exhibiting optical-grade contrast and capable of resolving the trabecular network in three dimensions through the cortical region with and without a soft-tissue overlayer are presented. The absolute nature and early demineralization-detection capability of a marker called thermal wave occupation index, estimated using the proposed modality, have been established. Selective imaging of regions of a specific mineral density range has been demonstrated in a mouse femur. The method is maximum-permissible-exposure compatible. In a matrix of bone and soft-tissue a depth range of ~3.8 mm has been achieved, which can be increased through instrumental and modulation waveform optimization. Furthermore, photoacoustic microscopy, a comparable modality with TC-PCT, has been used to resolve the trabecular structure and for comparison with the photothermal tomography.


Sonography and MRI of latissimus dorsi strain injury in four elite athletes.
The objective of this study was to describe the MR and sonographic findings in latissimus dorsi (LD) muscle strain in athletes, and to review the most common injuries described in the literature, most of which are humeral avulsions. Four injuries and two reinjuries of the myotendinous junction of the LD were followed from the day of injury until the return to play. Sonography (US) and MR imaging were performed in each case to confirm the diagnosis and to monitor the healing process. All cases had acute and isolated pain in the back of the shoulder while performing an eccentric maneuver of the arm and the shoulder. US and MR images demonstrated that injuries were located in the middle and cranial portion of the latissimus dorsi surrounding the myotendinous junction. After rehabilitation, all players played at high level again. Isolated lesions of LD are very rare. They can be demonstrated by US and MR images.


Dimethyl fumarate dosing in humans increases frataxin expression: A potential therapy for Friedreich's Ataxia.
Friedreich's Ataxia (FA) is an inherited neurodegenerative disorder resulting from decreased expression of the mitochondrial protein frataxin, for which there is no approved therapy. High throughput screening of clinically used drugs identified Dimethyl fumarate (DMF) as protective in FA patient cells. Here we demonstrate that DMF significantly increases frataxin gene (FXN) expression in FA cell model, FA mouse model and in DMF treated humans. DMF also rescues mitochondrial biogenesis deficiency in FA-patient derived cell model. We further examined the mechanism of DMF's frataxin induction in FA patient cells. It has been shown that transcription-inhibitory R-loops form at GAA expansion mutations, thus decreasing FXN expression. In FA patient cells, we demonstrate that DMF significantly increases transcription initiation. As a potential consequence, we observe significant reduction in both R-loop formation and transcriptional pausing thereby significantly increasing FXN expression. Lastly, DMF dosed Multiple Sclerosis (MS) patients showed significant increase in FXN expression by ~85%. Since inherited deficiency in FXN is the primary cause of FA, and DMF is demonstrated to increase FXN expression in humans, DMF could be considered for Friedreich's therapy.


Merkel Cell Polyomavirus Small T Antigen Induces Cancer and Embryonic Merkel Cell Proliferation in a Transgenic Mouse Model.
Merkel cell polyomavirus (MCV) causes the majority of human Merkel cell carcinomas (MCC) and encodes a small T (sT) antigen that transforms immortalized rodent fibroblasts in vitro. To develop a mouse model for MCV sT-induced carcinogenesis, we generated transgenic mice with a flox-stop-flox MCV sT sequence homologously recombined at the ROSA locus (ROSAsT), allowing Cre-mediated, conditional MCV sT expression. Standard tamoxifen (TMX) administration to adult UbcCreERT2; ROSAsT mice, in which Cre is ubiquitously expressed, resulted in MCV sT expression in multiple organs that was uniformly lethal within 5 days. Conversely, most adult UbcCreERT2; ROSAsT mice survived low-dose tamoxifen administration but developed ear lobe dermal hyperkeratosis and hypergranulosis. Simultaneous MCV sT expression and conditional homozygous p53 deletion generated multi-focal, poorly-differentiated, highly anaplastic tumors in the spleens and livers of mice after 60 days of TMX treatment. Mouse embryonic fibroblasts from these mice induced to express MCV sT exhibited anchorage-independent cell growth. To examine Merkel cell pathology, MCV sT expression was also induced during mid-embryogenesis in Merkel cells of Atoh1CreERT2/+; ROSAsT mice, which lead to significantly increased Merkel cell numbers in touch domes at late embryonic ages that normalized postnatally. Tamoxifen administration to adult Atoh1CreERT2/+; ROSAsT and Atoh1CreERT2/+; ROSAsT; p53flox/flox mice had no effects on Merkel cell numbers and did not induce tumor formation. Taken together, these results show that MCV sT stimulates progenitor Merkel cell proliferation in embryonic mice and is a bona fide viral oncoprotein that induces full cancer cell transformation in the p53-null setting.


Poly(ε-caprolactone)-banded spherulites and interaction with MC3T3-E1 cells.
We report that protein adsorption, cell attachment, and cell proliferation were enhanced on spherulites-roughened polymer surfaces. Banded spherulites with concentric alternating succession of ridges and valleys were observed on spin-coated thin films of poly(ε-caprolactone) (PCL) and two series of PCL binary homoblends composed of high- and low-molecular-weight components when they were isothermally crystallized at 25-52 °C. Their thermal properties, crystallization kinetics, and surface morphology were examined. The melting temperature (T(m)), crystallinity (χ(c)), crystallization rate, and spherulitic patterns showed strong dependence on the crystallization temperature (T(c)) and the blend composition. The surface roughness of the spherulites was higher when T(c) was higher; thus, the larger surface area formed in banded spherulites could adsorb more serum proteins from cell culture media. In vitro mouse preosteoblastic MC3T3-E1 cell attachment, proliferation, and nuclear localization were assessed on the hot-compressed flat disks and spherulites-roughened films of the high-molecular-weight PCL and one of its homoblends. The number of attached MC3T3-E1 cells and the proliferation rate were greater on the rougher surfaces than those on the flat ones. It is interesting to note that cell nuclei were preferentially, though not absolutely, located in or close to the valleys of the banded spherulites. The percentage of cell nuclei in the valleys was higher than 78% when the ridge height and adjacent ridge distance were ~350 and ~35 nm, respectively. This preference was weaker when the ridge height was lower or at a higher cell density. These results suggest that isothermal crystallization of semicrystalline polymers can be an effective thermal treatment method to achieve controllable surface roughness and pattern for regulating cell behaviors in tissue-engineering applications.


Statistical Paradigm for Organic Optoelectronic Devices: Normal Force Testing for Adhesion of Organic Photovoltaics and Organic Light-Emitting Diodes.
In this study, we assess the utility of a normal force (pull-test) approach to measuring adhesion in organic solar cells and organic light-emitting diodes. This approach is a simple and practical method of monitoring the impact of systematic changes in materials, processing conditions, or environmental exposure on interfacial strength and electrode delamination. The ease of measurement enables a statistical description with numerous samples, variant geometry, and minimal preparation. After examining over 70 samples, using the Weibull modulus and the characteristic breaking strength as metrics, we were able to successfully differentiate the adhesion values between 8-tris(hydroxyquinoline aluminum) (Alq3) and poly(3-hexyl-thiophene) and [6,6]-phenyl C61-butyric acid methyl ester (P3HT:PCBM) interfaces with Al and between two annealing times for the bulk heterojunction polymer blends. Additionally, the Weibull modulus, a relative measure of the range of flaw sizes at the fracture plane, can be correlated with the roughness of the organic surface. Finite element modeling of the delamination process suggests that the out-of-plane elastic modulus for Alq3 is lower than the reported in-plane elastic values. We suggest a statistical treatment of a large volume of tests be part of the standard protocol for investigating adhesion to accommodate the unavoidable variability in morphology and interfacial structure found in most organic devices.


Effects of 6-Week Supplementation with GliSODin on Parameters of Muscle Damages, Metabolic, and Work Performance at International Level Rowers after Specific Maximal Effort.
This study aimed to investigate the effect of supplementation with plant origin superoxide dismutase (SOD), GliSODin, on parameters of muscle damage, metabolic, and work performance at international level rowers. Twenty-eight rowers were included in a randomized, double-blind study. The study was conducted during a 6-week preparation period. At the beginning of the study and after 6 weeks of the supplementation period, all rowers were tested on a rowing ergometer. Blood samples were taken from the antecubital vein before and after every ergometer testing. Muscle damage markers creatine kinase (CK) and lactate dehydrogenase (LDH), total antioxidant capacity (TAC), inflammation parameters interleukin-6 (IL-6), and C-reactive protein (CRP) were measured. Rowing performance was assessed by lactate level in capillary blood and power output on the rowing ergometer. After supplementation, experimental group had significantly lower CK (p = 0.049) and IL-6 (p = 0.035) before and IL-6 (p = 0.050) after exhausting exercise on ergometer. Relative change of power output at 4 mmol/L concentration of lactate in blood, considering the initial and final test, was significantly higher (p = 0.020) in the supplemented group. It was concluded that GliSODin could be considered a good supplement in preventing some deleterious effects of intensive physical activity, including inflammation and muscle damage, and consequently, to enable a better rowing performance of elite rowers.


Presence of Intratumoral Calcifications and Vasculature Is Associated With Poor Overall Survival in Clear Cell Renal Cell Carcinoma.
The objective of this study was to explore the prognostic significance of the preoperative computed tomography (CT) features in clear cell renal cell carcinoma. The clinical data and CT data from 210 patients (1 grade 1, 84 grade 2, 92 grade 3, and 32 grade 4) generated with The Cancer Imaging Archive were reviewed. Overall survival was assessed using Kaplan-Meyer analysis. The relationship between CT features and survivals were evaluated using univariate and multivariable Cox regression analysis. The follow-up occurred between 13 and 3989 days (median, 1405 days; mean, 1434 days).On univariate Cox regressions, 4 preoperative CT features (intratumoral calcifications: yes vs no hazard ratio [HR], 2.054; 95% confidence interval [CI], 1.231-3.428; renal vein invasion: yes vs no HR, 2.013; 95% CI, 1.218-3.328; collecting system invasion: yes vs no HR, 2.139; 95% CI, 1.286-3.558; gross appearance of intratumoral vasculature: yes vs no HR, 2.385; 95% CI, 1.454-3.915) were significantly associated with overall survival (all P < 0.05). On multivariable Cox regression analysis, predictors of mortality in clear cell renal cell carcinoma were the presence of intratumoral calcifications (HR, 1.718; 95% CI, 1.014-2.911; P = 0.044) and gross appearance of intratumoral vasculature (HR, 2.137; 95% CI, 1.284-3.557; P = 0.003). Presence of intratumoral calcifications and vasculature can be potential prognostic features to screen patients for unfavorable prognosis.


Medical significance of Caenorhabditis elegans.
Caenorhabditis elegans is now the model organism of choice for a growing number of researchers. A combination of its apparent simplicity, exquisite genetics, the existence of a full molecular toolkit and a complete genome sequence makes it ideal for rapid and effective study of gene function. A survey of the C. elegans genome indicates that this 'simple' worm contains many genes with a high degree of similarity to human disease genes. For many human disease genes it has proven, and will continue to prove, difficult to elucidate their function by direct study. In such cases simpler model organisms may prove to be a more productive starting point. The basic function of a human disease gene may be studied in the background of C. elegans, in which the most important interactions are likely to be conserved, providing an insight into disease process in humans. Here we consider the significance of this modality for human disease processes and discuss how C. elegans may, in some cases, be ideal in the study of the function of human disease genes and act as a model for groups of parasitic nematodes that have a severe impact on world health.


Characterization of a right atrial subsidiary pacemaker and acceleration of the pacing rate by HCN over-expression.
Although the right atrium (RA contains subsidiary atrial pacemaker (SAP) tissue that can take over from the sinoatrial node (SAN) in sick sinus syndrome (SSS), SAP tissue is bradycardic. Little is known about SAP tissue and one aim of the study was to characterize ion channel expression to obtain insight into SAP pacemaker mechanisms. A second aim was to determine whether HCN over-expression (a 'biopacemaker'-like strategy) can accelerate the pacemaker rate producing a pacemaker that is similar in nature to the SAN. SAP tissue was isolated from the rat and the leading pacemaker site was characterized. Cell size at the leading pacemaker site in the SAP was smaller than in the RA and comparable to that in the SAN. mRNA levels showed the SAP to be similar to, but distinct from, the SAN. For example, in the SAN and SAP, expression of Tbx3 and HCN1 was higher and Nav1.5 and Cx43 lower than in the RA. Organ-cultured SAP tissue beat spontaneously, but at a slower rate than the SAN. Adenovirus-mediated gene transfer of HCN2 and the chimeric protein HCN212 significantly increased the pacemaker rate of the SAP close to that of the native SAN, but HCN4 was ineffective. SAP tissue near the inferior vena cava is bradycardic, but shares characteristics with the SAN. Pacing can be accelerated by the over-expression of HCN2 or HCN212. This provides proof of concept for the use of SAP tissue as a substrate for biopacemaking in the treatment of SSS.


Trends in the Incidence of Hospitalization for Major Diabetes-Related Complications in People With Type 1 and Type 2 Diabetes in Australia, 2010-2019.
To determine trends in the incidence of major diabetes-related complications in Australia. This study included 70,885 people with type 1 and 1,089,270 people with type 2 diabetes registered on the Australian diabetes registry followed from July 2010 to June 2019. Outcomes (hospitalization for myocardial infarction [MI], stroke, heart failure [HF], lower-extremity amputation [LEA], hypoglycemia, and hyperglycemia) were obtained via linkage to hospital admissions databases. Trends over time in the age-adjusted incidence of hospitalizations were analyzed using joinpoint regression and summarized as annual percent changes (APCs). In type 1 diabetes, the incidence of all complications remained stable, except for stroke, which increased from 2010-2011 to 2018-2019 (financial years; APC: +2.5% [95% CI 0.1, 4.8]), and hyperglycemia, which increased from 2010-2011 to 2016-2017 (APC: +2.7% [1.0, 4.5]). In type 2 diabetes, the incidence of stroke remained stable, while the incidence of MI decreased from 2012-2013 to 2018-2019 (APC: -1.7% [95% CI -2.8, -0.5]), as did the incidence of HF and hypoglycemia from 2010-2011 to 2018-2019 (APCs: -0.8% [-1.5, 0.0] and -5.3% [-6.7, -3.9], respectively); the incidence of LEA and hyperglycemia increased (APCs: +3.1% [1.9, 4.4], and +7.4% [5.9, 9.0]). Most trends were consistent by sex, but differed by age; in type 2 diabetes most improvements were confined to individuals aged ≥60 years. Trends in admissions for diabetes-related complications were largely stable in type 1 diabetes. In type 2 diabetes, hospitalization rates for MI, HF, and hypoglycemia fell over time, while increasing for LEA and hyperglycemia.


Associations of self-reported physical activity and depression in 10,000 Irish adults across harmonised datasets: a DEDIPAC-study.
Depression is a prevalent, debilitating, and often recurrent mood disorder for which successful first-line treatments remains limited. The purpose of this study was to investigate the cross-sectional associations between self-reported physical activity (PA) and depressive symptoms and status among Irish adults, using two existing datasets, The Irish Longitudinal Study on Ageing (TILDA) and The Mitchelstown Cohort Study. The two selected databases were pooled (n = 10,122), and relevant variables were harmonized. PA was measured using the short form International Physical Activity Questionnaire. Depressive symptoms were measured by the Center for Epidemiologic Studies Depression (CES-D) questionnaire. Participants were classified as meeting World Health Organization moderate-to-vigorous PA (MVPA) guidelines or not, and divided into tertiles based on weekly minutes of MVPA. A CES-D score of ≥16 indicated elevated depressive symptoms. Data collection were conducted in 2010-2011. Significantly higher depressive symptoms were reported by females (7.11 ± 7.87) than males (5.74 ± 6.86; p < 0.001). Following adjustment for age, sex, BMI, and dataset, meeting the PA guidelines was associated with 44.7% (95%CI: 35.0 to 52.9; p < 0.001) lower odds of elevated depressive symptoms. Compared to the low PA tertile, the middle and high PA tertiles were associated with 25.2% (95%CI: 8.7 to 38.6; p < 0.01) and 50.8% (95%CI: 40.7 to 59.2; p < 0.001) lower odds of elevated depressive symptoms, respectively. Meeting the PA guidelines is associated with lower odds of elevated depressive symptoms, and increased volumes of MVPA are associated with lower odds of elevated depressive symptoms.