Exercise Testing and Exercise Rehabilitation for Patients With Atrial Fibrillation.
Atrial fibrillation (AF) is a common cardiac arrhythmia associated with an increasing prevalence with advancing age. It is associated with dyspnea, exercise intolerance, and increased risk for clinical events, especially stroke and heart failure. This article provides a concise review of exercise testing and rehabilitation in patients with persistent or permanent AF. The first goal in the treatment of AF is to reduce symptoms (eg, palpitations) and a fast ventricular rate. The second goal is to reduce the risk of a stroke. Exercise testing and rehabilitation may be useful once these goals are achieved. However, there are no large, randomized exercise training trials involving patients with AF, and what data are available comes from single-site trials, secondary analyses, and observational studies. There are no specific indications for performing a graded exercise test in patients with AF; however, such testing may be used to screen for myocardial ischemia or evaluate chronotropic response during exertion. Among patients with AF, exercise capacity is 15% to 20% lower and peak heart rate is higher than in patients in sinus rhythm. Exercise rehabilitation improves exercise capacity, likely improves quality of life, and may improve symptoms associated with AF. Whole-body aerobic exercise is recommended. Atrial fibrillation is a common cardiac condition and in these patients, exercise rehabilitation favorably improves exercise capacity. However, prospective randomized controlled trials are needed to better define the effects of exercise training on safety; quality of life; clinical outcomes; and central, autonomic, and peripheral adaptations.


Refined structure for the complex of 1-deoxynojirimycin with glucoamylase from Aspergillus awamori var. X100 to 2.4-A resolution.
The three-dimensional structure of the complex of 1-deoxynojirimycin with glucoamylase II-(471) from Aspergillus awamori var. X100 has been determined to 2.4-A resolution. The model includes residues corresponding to residues 1-471 of glucoamylase I from Aspergillus niger, two molecules of bound 1-deoxynojirimycin and 605 sites for water molecules. The crystallographic R factor from refinement is 0.119, and the root-mean-squared deviation in bond distances is 0.012 A. The inhibitor complex confirms the location of the active site in the packing void of the alpha/alpha-barrel as proposed by Aleshin et al. [Aleshin, A., Golubev, A., Firsov, L., & Honzatko, R. B. (1992) J. Biol. Chem. 267, 19291-19298]. One inhibitor molecule is associated with strong electron density and represents the principal site of interaction of 1-deoxynojirimycin with the enzyme. The other 1-deoxynojirimycin molecule is associated with weak electron density and therefore, probably represents a binding site of low affinity. Interactions of 1-deoxynojirimycin with the enzyme at its principal site involve Arg 45, Asp 55, Arg 305, and carbonyl 177. In addition, a water molecule (water 500) hydrogen bonds to Glu 400 and the 6-hydroxyl of 1-deoxynojirimycin and is at an approximate distance of 3.3 A from the "anomeric" carbon of the inhibitor. The structural arrangement of functional groups near the inhibitor molecule suggests that Glu 179 is a catalytic acid, Glu 400 a catalytic base, and water 500 the attacking nucleophile in the hydrolysis of maltooligosaccharides. The relevance of the X-ray work to proposed mechanisms of enzymatic hydrolysis of oligosaccharides is discussed.


A randomized, double-blind, placebo-controlled trial of niacinamide for reduction of phosphorus in hemodialysis patients.
Niacinamide inhibits intestinal sodium/phosphorus transporters and reduces serum phosphorus in open-label studies. A prospective, randomized, double-blind, placebo-controlled crossover trial was performed for assessment of the safety and efficacy of niacinamide. Hemodialysis patients with phosphorus levels > or =5.0 mg/dl were randomly assigned to 8 wk of niacinamide or placebo, titrated from 500 to 1500 mg/d. After a 2-wk washout period, patients switched to 8 wk of the alternative therapy. Vitamin D analogs and calcimimetics were held constant; phosphorus binders were not changed unless safety criteria were met. Thirty-three patients successfully completed the trial. Serum phosphorus fell significantly from 6.26 to 5.47 mg/dl with niacinamide but not with placebo (5.85 to 5.98 mg/dl). A concurrent fall in calcium-phosphorus product was seen with niacinamide, whereas serum calcium, intact parathyroid hormone, uric acid, platelet, triglyceride, LDL, and total cholesterol levels remained stable in both arms. Serum HDL levels rose with niacinamide (50 to 61 mg/dl but not with placebo. Adverse effects were similar between both groups. Among patients who were > or =80% compliant, results were similar, although the decrease in serum phosphorus with niacinamide was more pronounced (6.45 to 5.28 mg/dl) and the increase in HDL approached significance (49 to 58 mg/dl). In hemodialysis patients, niacinamide effectively reduces serum phosphorus when co-administered with binders and results in a potentially advantageous increase in HDL cholesterol. Further study in larger randomized trials and other chronic kidney disease populations is indicated.


Strategies for weight maintenance in adult populations treated for overweight and obesity: a systematic review and meta-analysis.
Once weight loss is achieved, the challenge is to maintain this benefit. This review reports on the effectiveness of programs for weight-loss maintenance, as part of a larger review examining treatments for overweight and obese adults. We updated the search of a 2011 review on screening and management of overweight and obese adults. Four databases were searched. For inclusion, participants had to have lost weight in treatment and then been randomly assigned to a weight-maintenance intervention or control conditions. Studies from the 2011 review that met the criteria were included. Data were extracted and pooled (where possible) for outcomes related to weight-loss maintenance. Eight studies were included. Compared with control participants, intervention participants regained less weight (mean difference [MD] -1.44 kg, 95% confidence interval [CI] -2.42 to -0.47), regardless of whether the intervention was behavioural (MD-1.56 kg, 95% CI -3.10 to -0.02) or pharmacologic plus behavioural (MD -1.39 kg, 95% CI -2.86 to 0.08). Intervention participants also showed better weight maintenance than the control participants in terms of waist circumference (MD -2.30 cm, 95% CI -3.45 to -1.15) and body mass index (MD -0.95 kg/m(2), 95% CI -1.67 to -0.23). Participants undergoing pharmacologic plus behavioural interventions were more likely to maintain a loss of 5% or more of initial body weight than those in the control group (risk ratio [RR] 1.33, 95% CI 1.15 to 1.54); no difference was found for maintaining a weight loss of 10% or more (RR 1.76, 95% CI 0.75 to 4.12). Moderate quality evidence shows that overweight and obese adults can benefit from interventions for weight maintenance following weight loss. However, there is insufficient evidence on the long-term sustainability of these benefits. PROSPERO no. CRD42012002753.


Proteolysis of virulence regulator ToxR is associated with entry of Vibrio cholerae into a dormant state.
Vibrio cholerae O1 is a natural inhabitant of aquatic environments and causes the diarrheal disease, cholera. Two of its primary virulence regulators, TcpP and ToxR, are localized in the inner membrane. TcpP is encoded on the Vibrio Pathogenicity Island (VPI), a horizontally acquired mobile genetic element, and functions primarily in virulence gene regulation. TcpP has been shown to undergo regulated intramembrane proteolysis (RIP) in response to environmental conditions that are unfavorable for virulence gene expression. ToxR is encoded in the ancestral genome and is present in non-pathogenic strains of V. cholerae, indicating it has roles outside of the human host. In this study, we show that ToxR undergoes RIP in V. cholerae in response to nutrient limitation at alkaline pH, a condition that occurs during the stationary phase of growth. This process involves the site-2 protease RseP (YaeL), and is dependent upon the RpoE-mediated periplasmic stress response, as deletion mutants for the genes encoding these two proteins cannot proteolyze ToxR under nutrient limitation at alkaline pH. We determined that the loss of ToxR, genetically or by proteolysis, is associated with entry of V. cholerae into a dormant state in which the bacterium is normally found in the aquatic environment called viable but nonculturable (VBNC). Strains that can proteolyze ToxR, or do not encode it, lose culturability, experience a change in morphology associated with cells in VBNC, yet remain viable under nutrient limitation at alkaline pH. On the other hand, mutant strains that cannot proteolyze ToxR remain culturable and maintain the morphology of cells in an active state of growth. Overall, our findings provide a link between the proteolysis of a virulence regulator and the entry of a pathogen into an environmentally persistent state.


Toward standardizing deuterium content reporting in hydrogen exchange-MS.
We introduce a method to monitor dispensing ratios during labeling reactions in hydrogen exchange (HX)-MS. The method corrects for systematic and random dispensing errors and harmonizes data incorporating variable %D2O in the experiment design. A correction factor for deuterium levels is obtained by quantifying the relative signal intensities arising from nonexchanging heavy caffeine (spiked into labeling buffer) and light caffeine (spiked into sample solutions). Dispensing variability over a wide range of %D2O composition can be detected and corrected to a common value, and although random dispensing error is usually minor, we show it can be the limiting factor in high quality signal measurements. Applying a dispensing control is therefore an effective tool for monitoring measurement precision in HX-MS.


What causes acute coronary syndromes? Applying Koch's postulates.
The term "acute coronary syndromes" (ACS) is used to describe a heterogeneous spectrum of clinical conditions. This includes myocardial infarction, non-ST-elevation myocardial infarction, and unstable angina. These conditions are linked by a similar constellation of signs and symptoms but not necessarily by a common pathophysiology. They are syndromes. Several different hypotheses exist that have attempted to explain the pathological mechanisms that are involved in these conditions, however, it is not clear whether ACS are caused by variations of a single disease process or by several disease processes. The contribution of both vessel wall- and blood-related factors in the pathogenesis of acute coronary syndromes is herein discussed with the guidance of Koch's postulates.


Understanding the electronic and phonon transport properties of a thermoelectric material BiCuSeO: a first-principles study.
Using the first-principles pseudopotential method and Boltzmann transport theory, we give a comprehensive understanding of the electronic and phonon transport properties of the thermoelectric material BiCuSeO. By choosing an appropriate hybrid functional for the exchange-correlation energy, we find that the system is a semiconductor with a direct band gap of ∼0.8 eV, which is quite different from those obtained previously using standard functionals. Detailed analysis of a three-dimensional energy band structure indicates that there is a valley degeneracy of eight around the valence band maximum, which leads to a sharp density of states and is responsible for a large p-type Seebeck coefficient. Moreover, we find that the density of states effective mass is much larger and results in a very low hole mobility for BiCuSeO. On the other hand, we discover two flat phonon branches contributed by the Cu and Se atoms, which can effectively block heat transfer. Combined with large atomic displacement parameters of the Cu atom, we believe that the intrinsically low lattice thermal conductivity in BiCuSeO is mainly caused by the Cu atoms, instead of the prevailingly believed Bi atoms. The thermoelectric figure-of-merit is also predicted and compared with available experimental results.


Five-week dietary exposure to dry diets alters the faecal bacterial populations in the domestic cat (Felis catus).
The effects of wet (canned) or dry (kibbled) diets on faecal bacterial populations in the cat were investigated in eight domestic short-haired cats (four males and four females; averaging 6 years of age and 3.4 kg) in a nested design. The cats were fed ad libitum a commercially available wet diet (moisture 82.0 %, crude protein 51.7 %, fat 28.9 %, carbohydrate (CHO) 8.9 % and ash 10.6 % DM) for 5 weeks. On the fifth week, individual feed intakes and faecal outputs were determined. Fresh faecal samples were collected twice daily, mixed for homogeneity, subsampled and stored at - 85 °C until analysis. The cats were then switched to a commercially available dry diet (moisture 8.5 %, crude protein 33.0 %, fat 11.0 %, CHO 49.4 % and ash 6.6 % DM) for 5 weeks, and fresh faeces were sampled as described previously. Energy intake tended to be higher in cats fed dry diets (P < 0.10), but body weight was similar between the two feeding periods (P>0.05). Denaturing gradient gel electrophoresis (DGGE) of bacterial 16S rRNA genes amplified from DNA extracted from faeces was performed. The unweighted pair group method with arithmetic mean cluster analysis of bacterial community profiles using Pearson's correlation revealed diet-specific clustering when the same cats were fed on either a dry or a wet diet (dissimilarity between the groups, 88.6 %; P < 0.001). Subsequent cloning and sequencing of five selected distinct DGGE bands indicated that members of the Pelomonas and Fusobacteriaceae were influenced by a short-term change in diet format. This suggests that 5-week dietary exposure is sufficient to alter gastrointestinal microflora.


Hepatocyte nuclear factor 1alpha gene inactivation impairs chromatin remodeling and demethylation of the phenylalanine hydroxylase gene.
Hepatocyte nuclear factor 1 alpha (HNF1alpha) is a homeoprotein that is expressed in the liver, kidney, pancreas, and digestive tract. Its inactivation in mouse resulted in decreased transcription of known target genes such as albumin and alpha1-antitrypsin. In contrast, the phenylalanine hydroxylase (PAH) gene was totally silent and unresponsive to normal inducers like glucocorticoids and cyclic AMP in the liver. DNase I and micrococcal nuclease digestion of liver nuclei showed that HNF1alpha inactivation had drastic effects on the chromatin structure of the PAH regulatory regions. Three DNase I-hypersensitive sites (HSSI, HSSII, and HSSIII), typical of the actively transcribed PAH gene, were undetectable in liver from HNF1alpha-deficient animals. Both HSSII and HSSIII elements harbor HNF1 sites, but only the latter has detectable enhancer activity in transient-transfection assays. In addition, the PAH promoter in livers of HNF1alpha-deficient animals was methylated. These results suggest that HNF1alpha could activate transcription through two mechanisms. One implies participation in the recruitment of the general transcription machinery to the promoter, and the second involves the remodeling of chromatin structure and demethylation that would allow transcription factors to interact with their cognate cis-acting elements.


Characterization of T helper cell responses to the trypanosome variant surface glycoprotein.
T cell responses to the variant surface glycoprotein (VSG) previously have not been detected in animals infected with the African trypanosomes despite the fact that such animals make strong T-dependent B cell responses to VSG molecules displayed by the parasites. In the present study, we have examined B 10.BR mice for VSG-specific Th cell responses at different times after infection with Trypanosoma brucei rhodesiense clone LouTat 1. T cell populations derived from different tissues were tested for their ability to proliferate and secrete cytokines when stimulated with purified LouTat 1 VSG. Furthermore, VSG-specific T cell lines and clones were derived from immunized mice and examined for their phenotypic and functional profiles in comparison with T cell responses of infected mice. The results of this study show that VSG-specific T cells were not consistently detected in the peripheral lymphoid tissues such as spleen or lymph nodes of infected animals. In contrast, VSG Ag-specific T cells were detectable principally in the peritoneal T cell populations of infected mice. Peritoneal T cells did not proliferate in response to VSG, yet produced substantial cytokine responses when stimulated; the cytokines produced were IFN-gamma and IL-2, without detectable IL-4. The cellular phenotype of VSG-responsive T cells was that of classical Th cells in that all cells were CD4-positive and expressed the CD3 alpha/beta TCR membrane complex. Thus, the VSG appears to preferentially stimulate a Th1 cell subset response during infection. Intrinsic molecular characteristics of the VSG molecule did not induce mice to make this response, however, since VSG-specific T cell lines derived from VSG-immunized mice displayed cytokine profiles characteristic of both Th1 and Th2 cells. Isolation of Th1 clones from selected lines demonstrated that these cells displayed the same membrane-phenotypic characteristics and cytokine profiles as the T cells from infected mice. Furthermore, all Th clones were VSG type-specific, APC-dependent, and I-Ak-restricted in their responses. In summary, these experiments provide the first direct evidence for VSG-specific responses at the T cell level. T cell responses to the VSG molecule during infection appear to be anatomically compartmentalized and exhibit evidence of clonal maturation (cytokine production) but not clonal expansion (proliferation) after antigenic stimulation. The cellular phenotype and cytokine profiles predict that infection predisposes the animals to mount Th1 cell subset responses to VSG. The results of this study, including the T clones generated, provide an experimental basis for examining the regulation of VSG-specific immune responses during infection.


Safe Drive Stay Alive: exploring effectiveness of a real-world driving intervention for predrivers and the utility of the health action process approach.
Young drivers are greatly over-represented in road traffic collisions (RTCs) worldwide. Interventions attempt to change driving-related behaviours to reduce injuries and deaths from RTCs. The current study evaluated the effectiveness of the well-established Fife Safe Drive Stay Alive (SDSA) practice-based intervention on determinants of driving behaviour using the health action process approach (HAPA) model. Adolescent participants (predominantly predrivers) attending the SDSA intervention from schools and colleges in Fife, Scotland, were invited to complete an evaluation at baseline and at 3 months exploring motivational determinants of driving behaviour (eg, risk perception). Intervention content was examined for behaviour change techniques (BCTs). Eighty-seven participants completed both baseline and follow-up evaluations. The motivational HAPA model variables predicted driving intentions. There was no significant overall effect of the SDSA intervention between baseline and 3-month follow-up. Seven negatively framed BCTs were used in the intervention. The effectiveness of SDSA is questioned; however, the study supports the use of the HAPA model in explaining driving intentions, and therefore, may usefully inform driving interventions.


Genetic architecture of ambulatory blood pressure in the general population: insights from cardiovascular gene-centric array.
Genetic determinants of blood pressure are poorly defined. We undertook a large-scale, gene-centric analysis to identify loci and pathways associated with ambulatory systolic and diastolic blood pressure. We measured 24-hour ambulatory blood pressure in 2020 individuals from 520 white European nuclear families (the Genetic Regulation of Arterial Pressure of Humans in the Community Study) and genotyped their DNA using the Illumina HumanCVD BeadChip array, which contains ≈ 50 000 single nucleotide polymorphisms in >2000 cardiovascular candidate loci. We found a strong association between rs13306560 polymorphism in the promoter region of MTHFR and CLCN6 and mean 24-hour diastolic blood pressure; each minor allele copy of rs13306560 was associated with 2.6 mm Hg lower mean 24-hour diastolic blood pressure (P = 1.2 × 10⁻⁸). rs13306560 was also associated with clinic diastolic blood pressure in a combined analysis of 8129 subjects from the Genetic Regulation of Arterial Pressure of Humans in the Community Study, the CoLaus Study, and the Silesian Cardiovascular Study (P=5.4 × 10⁻⁶). Additional analysis of associations between variants in gene ontology-defined pathways and mean 24-hour blood pressure in the Genetic Regulation of Arterial Pressure of Humans in the Community Study showed that cell survival control signaling cascades could play a role in blood pressure regulation. There was also a significant overrepresentation of rare variants (minor allele frequency: < 0.05) among polymorphisms showing at least nominal association with mean 24-hour blood pressure indicating that a considerable proportion of its heritability may be explained by uncommon alleles. Through a large-scale gene-centric analysis of ambulatory blood pressure, we identified an association of a novel variant at the MTHFR/CLNC6 locus with diastolic blood pressure and provided new insights into the genetic architecture of blood pressure.


Quantitative evaluation of noise reduction strategies in dual-energy imaging.
In this paper we describe a quantitative evaluation of the performance of three dual-energy noise reduction algorithms: Kalender's correlated noise reduction (KCNR), noise clipping (NOC), and edge-predictive adaptive smoothing (EPAS). These algorithms were compared to a simple smoothing filter approach, using the variance and noise power spectrum measurements of the residual noise in dual-energy images acquired with an a-Si TFT flat-panel x-ray detector. An estimate of the true noise was made through a new method with subpixel accuracy by subtracting an individual image from an ensemble average image. The results indicate that in the lung regions of the tissue image, all three algorithms reduced the noise by similar percentages at high spatial frequencies (KCNR=88%, NOC=88%, EPAS=84%, NOC/KCNR=88%) and somewhat less at low spatial frequencies (KCNR=45%, NOC=54%, EPAS=52%, NOC/KCNR=55%). At low frequencies, the presence of edge artifacts from KCNR made the performance worse, thus NOC or NOC combined with KCNR performed best. At high frequencies, KCNR performed best in the bone image, yet NOC performed best in the tissue image. Noise reduction strategies in dual-energy imaging can be effective and should focus on blending various algorithms depending on anatomical locations.


D-6-Deoxy-myo-inositol 1,3,4,5-tetrakisphosphate, a mimic of d-myo-inositol 1,3,4,5-tetrakisphosphate: biological activity and pH-dependent conformational properties.
D-6-Deoxy-myo-inositol 1,3,4,5-tetrakisphosphate [D-6-deoxy-Ins(1,3,4,5)P(4)] 3 is a novel deoxygenated analogue of D-myo-inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P(4)] 2, a central and enigmatic molecule in the polyphosphoinositide pathway of cellular signalling. D-6-Deoxy-Ins(1,3,4,5)P(4) is a moderate inhibitor of Ins(1,4,5)P(3) 5-phosphatase [1.8microM] compared to Ins(1,3,4,5)P(4) [0.15microM] and similar to that of L-Ins(1,3,4,5)P(4) [1.8microM]. In displacement of [(3)H] Ins(1,4,5)P(3) from the rat cerebellar Ins(1,4,5)P(3) receptor, while slightly weaker [IC(50)=800nM] than that of D-Ins(1,3,4,5)P(4) [IC(50)=220nM], 3 is less markedly different and again similar to that of L-Ins(1,3,4,5)P(4) [IC(50)=660nM]. 3 is an activator of I(CRAC) when inward currents are measured in RBL-2H3-M1 cells using patch-clamp electrophysiological techniques with a facilitation curve different to that of Ins(1,3,4,5)P(4). Physicochemical properties were studied by potentiometric (31)P and (1)H NMR titrations and were similar to those of Ins(1,3,4,5)P(4) apart from the observation of a biphasic titration curve for the P1 phosphate group. A novel vicinal phosphate charge-induced conformational change of the inositol ring above pH 10 was observed for D-6-deoxy-Ins(1,3,4,5)P(4) that would normally be hindered because of the central stabilising role played by the 6-OH group in Ins(1,3,4,5)P(4). We conclude that the 6-OH group in Ins(1,3,4,5)P(4) is crucial for its physicochemical behaviour and biological properties of this key inositol phosphate.


Non-cross Bridge Viscoelastic Elements Contribute to Muscle Force and Work During Stretch-Shortening Cycles: Evidence From Whole Muscles and Permeabilized Fibers.
The sliding filament-swinging cross bridge theory of skeletal muscle contraction provides a reasonable description of muscle properties during isometric contractions at or near maximum isometric force. However, it fails to predict muscle force during dynamic length changes, implying that the model is not complete. Mounting evidence suggests that, along with cross bridges, a Ca2+-sensitive viscoelastic element, likely the titin protein, contributes to muscle force and work. The purpose of this study was to develop a multi-level approach deploying stretch-shortening cycles (SSCs) to test the hypothesis that, along with cross bridges, Ca2+-sensitive viscoelastic elements in sarcomeres contribute to force and work. Using whole soleus muscles from wild type and mdm mice, which carry a small deletion in the N2A region of titin, we measured the activation- and phase-dependence of enhanced force and work during SSCs with and without doublet stimuli. In wild type muscles, a doublet stimulus led to an increase in peak force and work per cycle, with the largest effects occurring for stimulation during the lengthening phase of SSCs. In contrast, mdm muscles showed neither doublet potentiation features, nor phase-dependence of activation. To further distinguish the contributions of cross bridge and non-cross bridge elements, we performed SSCs on permeabilized psoas fiber bundles activated to different levels using either [Ca2+] or [Ca2+] plus the myosin inhibitor 2,3-butanedione monoxime (BDM). Across activation levels ranging from 15 to 100% of maximum isometric force, peak force, and work per cycle were enhanced for fibers in [Ca2+] plus BDM compared to [Ca2+] alone at a corresponding activation level, suggesting a contribution from Ca2+-sensitive, non-cross bridge, viscoelastic elements. Taken together, our results suggest that a tunable viscoelastic element such as titin contributes to: (1) persistence of force at low [Ca2+] in doublet potentiation; (2) phase- and length-dependence of doublet potentiation observed in wild type muscles and the absence of these effects in mdm muscles; and (3) increased peak force and work per cycle in SSCs. We conclude that non-cross bridge viscoelastic elements, likely titin, contribute substantially to muscle force and work, as well as the phase-dependence of these quantities, during dynamic length changes.


Cultural Competence Assessment Instrument: Initial Italian validation and proposed refinement.
Italy has become a target of immigration in the last three decades. Accordingly, the Italian population is progressively changing, becoming increasingly culturally different. Cultural competences are a fundamental requirement for many industries and, especially, for healthcare organizations. The aim of this paper is to propose an initial Italian validation of the Cultural Competence Assessment Instrument (CCAI) and to propose a refinement of this scale in terms of measured constructs. The CCAI was translated into Italian through a team-based iterative approach and then administered to a sample of 289 nurses with symbolic and realistic threat scale and social dominance orientation scale. An on-line cross-sectional survey questionnaire was used. Confirmatory factor analysis revealed that the original two dimensions of the CCAI can be divided into two other sub-scales, thus leaving us with the following dimensions: cultural awareness, cultural sensitivity, seeking information and active behavior. These dimensions appeared to be sufficiently reliable and independent one from another. Moreover, they showed specific and different correlations with other measured constructs. The Italian version of the CCAI would seem to be a useful instrument for measuring both attitudes and behavioral intention of nurses with respect to intercultural care. Using four dimensions instead of two appears to increase the understanding of professionals' cultural competence and supply a deeper picture of dimensions which compose cultural competence in healthcare settings.


Attendance and diagnostic yield of repeated two-sample faecal immunochemical test screening for colorectal cancer.
Limited data exist on attendance and additional yield of 2-sample faecal immunochemical testing (FIT) screening during multiple rounds. We therefore conducted a population-based colorectal cancer screening trial comparing attendance and yield of repeated 1-sample and 2-sample FIT screenings. Two randomly selected groups of average-risk subjects aged 50-74 years were invited for two rounds of either 1-sample (n=5007) or 2-sample (n=3197) FIT (OC-sensor Micro) screening. The test was considered positive if at least one sample was positive (cut-off 50 ng/mL; 10 µg haemoglobin/g). The cumulative attendance rate was similar for repeated 1-sample and 2-sample FIT screenings (1-sample FIT: 68.1%; 2-sample FIT: 67.1%, p=0.368). The positivity rate in the second round was lower for 1-sample FIT (6.2%, 95% CI 5.4% to 7.2%) than for 2-sample FIT (8.4%, 95% CI 7.1% to 9.8%, p=0.007), whereas the detection rate of advanced neoplasia (AN, 1-sample FIT: 1.9%, 95% CI 1.2% to 2.2%; 2-sample FIT: 1.7%, 95% CI 1.2% to 2.5%, p=0.861) and the positive predictive value (1-sample FIT: 32%, 95% CI 24% to 40%; 2-sample FIT: 21%, 95% CI 15% to 29%, p=0.075) did not differ. After two rounds of screening, the cumulative diagnostic yield of AN for 1-sample FIT was 29.3 per 1000 invitees, compared with 34.0 for 2-sample FIT (p=0.241). Using 2-sample FIT instead of 1-sample FIT does not result in a higher detection rate of AN in the second round of repeated FIT screening. Furthermore, both strategies lead to a similar yield of AN over two rounds. These findings imply that 1-sample FIT screening is preferred over 2-sample FIT screening.


Association between duration of electronic screen use for non-educational purposes and depression symptoms among middle and high school students: a cross-sectional study in Zhejiang Province, China.
Existing literature on the association of electronic screen use duration with depression among adolescents is contradictory. The current study aimed to elucidate the association between duration of electronic screen use for non-educational purposes and depression symptoms among middle and high school students in Zhejiang Province, China. A cross-sectional study of 27,070 students in grades 7-12 from 376 middle and high schools was conducted through an anonymous self-administered questionnaire between April and June 2022. Poisson regression was utilized to examine the association between electronic screen use duration for non-educational purposes and depression symptoms. Of the 27,006 eligible students, 51.6% (13932) were boys and the mean (SD) age was 15.6(1.7) years. The overall prevalence of symptoms of depression was 22.4% (95%CI 21.4-23.4); girls (27.6%, 26.2-29.0) had a higher prevalence than boys (17.7%, 16.7-18.8). After adjustment for socio-demographic status, lifestyle factors, self-perceived health, academic performance, loneliness and sadness, compared to those who did not use electronic screens for non-educational purposes, the prevalence ratios (PRs) for depression symptoms were 1.03 (95% CI 1.02-1.04) for those exposed to electronic screens for <1 h/day, 1.07 (1.05-1.09) for 1.0-1.9 h/day, 1.10 (1.07-1.13) for 2.0-2.9 h/day, 1.14 (1.10-1.18) for 3.0-3.9 h/day, 1.18 (1.12-1.23) for 4.0-4.9 h/day, and 1.21 (1.15-1.29) for ≥5 h/day. Duration of electronic screen use for non-educational purposes was positively associated with symptoms of depression among middle and high school students, even with a relatively short daily duration of use.


High prevalence of central nervous system medications in community-dwelling older adults with dementia over a three-year period.
Few recent studies have investigated the prevalence and outcomes for central nervous system (CNS)-active medication use in older persons with dementia (PWD) who live in the community. Thus, the purpose of this study was to describe the health outcomes and patterns of use of CNS-active drugs in PWD living in the community. Using a retrospective study design from a southeastern managed care organization (MCO), claims data were collected for three years on all identified cases with dementia and included age, gender, medical diagnoses for each claim (International Classification of Disease [ICD-9 code]) and prescription drugs (National Drug Code [NDC]). Individuals (N = 960) were selected who were continuously enrolled and had prescription drug coverage. Over 79% of PWD in this sample were on a CNS-active medication during the three-year period and 35% were on a benzodiazepine. The highest number of drug-related problems (DRPs) within 45 days after receiving a CNS drug prescription were for syncope, fatigue, altered level of consciousness, delirium, constipation, falls and fractures. This study illustrates the need to further examine inappropriate CNS-active medication use in PWD and to test non-pharmacologic therapies for the clinical problems that initiate their use in PWD.


Diagnosis and treatment of myocardial fibrosis in hypertensive heart disease.
Although hypertensive heart disease (HHD) is clinically characterized by development of left ventricular hypertrophy in the absence of a cause other than arterial hypertension, changes in the composition of myocardial tissue also develop in arterial hypertension, leading to structural remodeling of the myocardium (eg, fibrosis). Myocardial fibrosis is the major determinant of diastolic dysfunction/failure in patients with HHD. Recent available data on the determination of serum concentrations of collagen-derived serum peptides, as well as quantitative analysis of echoreflectivity to address the presence of fibrosis in the myocardium of hypertensive patients, are promising. In addition, preliminary data suggest that the goal of reducing myocardial fibrosis is achievable using specific pharmacological agents in patients with HHD.


Solid-spaced filters: an alternative for narrow-bandpass applications.
Solid-spaced filters are composed of one or several thin wafers of excellent optical quality acting as Fabry-Perot spacer layers. We study the different steps of the design and the manufacture of filters following dense-wavelength-division-multiplexing specifications. The design method of such filters requires a tight synergy between numerical simulations and experimental characterizations to correct possible thickness errors. Experimental results of the manufacture and characterization of a three-cavity narrow-bandpass filter and of an interleaver filter are given.


Motivation for Critical Thinking in Nursing Students in Chile.
Nursing education includes a constant challenge regarding the competencies that nurses must possess to provide high-quality nursing care. One of these competencies is critical thinking (CT), and motivation for critical thinking (MCT) has been identified as an element necessary to develop CT. Expectancy and value are important components of the MCT. This study aimed to analyze sociodemographic and academic factors related to MCT in Chilean nursing students. This study is an analytical-correlational, cross-sectional study conducted with 478 nursing students in Chile. The data were collected using an online questionnaire. The Critical Thinking Motivation Scale was used to measure MCT. Pearson's square, Chi-square, means comparison test, and binomial logistic regression were used to analyze the data. The expectancy component of the MCT had the lowest scores on the scale. Age was associated with the cost subcomponent (p < .03), and high school was associated with the interest subcomponent (p < .01). Academic factors contributing to MCT were problem-based learning, professor motivation, and the inclusion of MCT in nursing disciplinary subjects. Strategies that improve CT and MCT should be implemented and evaluated periodically from the beginning of a nursing career. Faculty should be trained to motivate students to think critically.


Speech-Evoked Auditory Brainstem Response in Individuals with Diabetes Mellitus Type 2.
Diabetes is the most common glucose level dependent metabolic disorder and studies have shown that hearing impairment can be a long-term subclinical complication. Studies to investigate auditory system involvement in diabetes has focused majorly on the auditory brainstem response (ABR), otoacoustic emission, and basic audiological measures. Hence in the current study, we used speech-evoked ABR (S-ABR) as a tool to see the effect of diabetes on both a transient and sustained response of the auditory brainstem to a conventionally used consonant-vowel (CV) stimuli /da/. This preliminary investigation was done on 22 individuals in the age range of 40-55 years. 11 individuals were diabetics for a minimum period of five years. The S-ABR was recorded for all the participants with speech stimuli /da/ of 40 ms duration. Latency analysis of the waves V, A, D, E, F, and O were carried out. The statistical analysis included descriptive measures, paired t tests, and MANOVA. The findings of the current study suggest that middle-aged individuals with diabetes have a significant deficiency in auditory processing at the brainstem level. Both transient (wave V (p=0.00), A (p=0.00), and O (p=0.00)) and sustained responses (wave D (p=0.001), E (p=0.00), and F (p=0.00)) of the S-ABR were found to be affected in diabetic individuals compared to age-matched non-diabetic individuals. Considering diabetes is a common metabolic disorder in the middle-aged Indian population, the findings of the present study can have significant clinical implication.


Cognitive Recovery After Stroke: Memory.
Memory impairment occurs in over a third of patients after symptomatic stroke. Memory deficits rarely occur in isolation but are an important component of the poststroke cognitive syndrome because of the strong relationship with the risk of poststroke dementia. In this review, we summarize available data on impairment of episodic memory, with a particular emphasis on the natural history of memory impairment after stroke and the factors influencing trajectory informed by an updated systematic review. We next discuss the pathophysiology of memory impairment and mechanisms of both decline and recovery of function. We then turn to the practical issue of measurement of memory deficits after stroke, emerging biomarkers, and therapeutic approaches. Our review identifies critical gaps, particularly in studies of the natural history that properly map the long-term trajectory of memory and the associations with factors that modulate prognosis. Few studies have used advanced neuroimaging and this, in conjunction with other biomarker approaches, has the potential to provide a much richer understanding of the mechanisms at play and promising therapeutic avenues.


Discriminant brain connectivity patterns of performance monitoring at average and single-trial levels.
Electrophysiological and neuroimaging evidence suggest the existence of common mechanisms for monitoring erroneous events, independent of the source of errors. Previous works have described modulations of theta activity in the medial frontal cortex elicited by either self-generated errors or erroneous feedback. In turn, similar patterns have recently been reported to appear after the observation of external errors. We report cross-regional interactions after observation of errors at both average and single-trial levels. We recorded scalp electroencephalography (EEG) signals from 15 subjects while monitoring the movement of a cursor on a computer screen. Connectivity patterns, estimated using multivariate auto-regressive models, show increased error-related modulations of the information transfer in the theta and alpha bands between frontocentral and frontolateral areas. Conversely, a decrease of connectivity in the beta band is also observed. These network patterns are similar to those elicited by self-generated errors. However, since no motor response is required, they appear to be related to intrinsic mechanisms of error processing, instead of being linked to co-activation of motor areas. Noticeably, we demonstrate that cross-regional interaction patterns can be estimated on a trial-by-trial basis. These trial-specific patterns, consistent with the multi-trial analysis, convey discriminant information on whether a trial was elicited by observation of an erroneous action. Overall, our study supports the role of frequency-specific modulations in the medial frontal cortex in coordinating cross-regional activity during cognitive monitoring at a single-trial basis.


Immediate provisionalization of NanoTite implants in support of single-tooth and unilateral restorations: one-year interim report of a prospective, multicenter study.
Clinical studies reporting immediate loading of endosseous implants for edentulous cases and for fixed partial restorations have been well documented with satisfactory survival rates. Implants with a recently developed, nanometer-scale surface topography (NanoTite, BIOMET 3i, Palm Beach Gardens, FL, USA), created by discrete crystalline depositions (DCD) of calcium phosphate nano-crystals onto a dual acid-etched (DAE) surface, show enhanced early fixation in preclinical studies when compared with DAE-surfaced implants. These outcomes suggest DCD-surfaced implants may be advantageous for immediate loading approaches. The aim of this prospective, multicenter, observational study is to report clinical outcomes for DCD-surfaced implants placed in immediate functional support of single- and multi-unit restorations according to an immediate loading protocol. One hundred eighty-five patients enrolled at 15 international study centers received a total of 335 implants supporting 216 immediate provisionalizations consisting of 128 single-tooth restorations and 88 fixed restorations. Of the 335 implants, 77% are located in posterior and 23% in anterior regions with 55.5% of the total in mandibles and 44.5% in maxillae. Patients were evaluated for implant mobility, gingival health, symptomatology, and radiographic outcomes. At the time of this 1-year interim report, a total of 17 failures have been observed in 11 patients, yielding a cumulative survival rate of 94.9%. Relative to other prospective, multicenter studies of immediately loaded implants with various surface enhancements, NanoTite implants perform comparatively well when immediately provisionalized with single-tooth and fixed restorations.


Plasma microRNAs as biomarkers of pancreatic cancer risk in a prospective cohort study.
Noninvasive biomarkers for early pancreatic ductal adenocarcinoma (PDAC) diagnosis and disease risk stratification are greatly needed. We conducted a nested case-control study within the Prospective Investigation into Cancer and Nutrition (EPIC) cohort to evaluate prediagnostic microRNAs (miRs) as biomarkers of subsequent PDAC risk. A panel of eight miRs (miR-10a, -10b, -21-3p, -21-5p, -30c, -106b, -155 and -212) based on previous evidence from our group was evaluated in 225 microscopically confirmed PDAC cases and 225 controls matched on center, sex, fasting status and age/date/time of blood collection. MiR levels in prediagnostic plasma samples were determined by quantitative RT-PCR. Logistic regression was used to model levels and PDAC risk, adjusting for covariates and to estimate area under the receiver operating characteristic curves (AUC). Plasma miR-10b, -21-5p, -30c and -106b levels were significantly higher in cases diagnosed within 2 years of blood collection compared to matched controls (all p-values <0.04). Based on adjusted logistic regression models, levels for six miRs (miR-10a, -10b, -21-5p, -30c, -155 and -212) overall, and for four miRs (-10a, -10b, -21-5p and -30c) at shorter follow-up time between blood collection and diagnosis (≤5 yr, ≤2 yr), were statistically significantly associated with risk. A score based on the panel showed a linear dose-response trend with risk (p-value = 0.0006). For shorter follow-up (≤5 yr), AUC for the score was 0.73, and for individual miRs ranged from 0.73 (miR-212) to 0.79 (miR-21-5p).


Does concomitant cholecystectomy at time of Roux-en-Y gastric bypass impact adverse operative outcomes?
We hypothesized that patients undergoing Roux-en-Y gastric bypass (RYGB) with concomitant cholecystectomy (RYGB + C) would be at greater risk for adverse events compared to patients undergoing RYGB alone. Patients who underwent a RYGB were identified in the 2005-2009 American College of Surgeons National Surgical Quality Improvement Program Database. Multivariate logistic regression with adjustment for confounding variables was utilized to identify risk factors for mortality at 30 days, major adverse events, and prolonged length of stay (PLOS). We identified 32,946 patients who underwent RYGB; of these, 1,731 (5.2%) underwent RYGB + C. Overall, RYGB + C was a risk factor for predicting major adverse events following laparoscopic but not open procedures. Regardless of approach, PLOS was more common among RYGB + C patients following adjustment. Overall mortality at 30 days was low and did not vary with concomitant cholecystectomy following adjustment. The risk for major adverse events is significantly greater for RYGB + C patients following laparoscopic procedures, and the risk for PLOS is greater for RYGB + C patients following both open and laparoscopic procedures. The short-term risks identified in this study can assist in decision-making when considering concomitant cholecystectomy at the time of RYGB.


Parks promoting physical activity: synthesis of findings from interventions in seven national parks.
We synthesized the results of 7 National Park Service pilot interventions designed to increase awareness of the health benefits from participation in recreation at national parks and to increase physical activity by park visitors. A content analysis was conducted of the final evaluation reports of the 7 participating parks. Pooled data were also analyzed from a standardized trail-intercept survey administered in 3 parks. The theme of new and diverse partnerships was the most common benefit reported across the 7 sites. The 2 parks that focused on youth showed evidence of an increase in awareness of the benefits of physical activity. Many of the other sites found high levels of awareness at baseline (approaching 90%), suggesting little room for improvement. Five of the 7 projects showed evidence of an increase in physical activity that was associated with the intervention activities. Multivariate analyses suggested that the media exposure contributed to a small but significant increase in awareness of the importance of physical activity (6%) and number of active visits (7%). Enhancements and replication of these programs represents a promising opportunity for improving partnerships between public health and recreation to increase physical activity.


Characterization of three ammonium transporters of the glomeromycotan fungus Geosiphon pyriformis.
Members of the Glomeromycota form the arbuscular mycorrhiza (AM) symbiosis. They supply plants with inorganic nutrients, including nitrogen, from the soil. To gain insight into transporters potentially facilitating nitrogen transport processes, ammonium transporters (AMTs) of Geosiphon pyriformis, a glomeromycotan fungus forming a symbiosis with cyanobacteria, were studied. Three AMT genes were identified, and all three were expressed in the symbiotic stage. The localization and functional characterization of the proteins in a heterologous yeast system revealed distinct characteristics for each of them. AMT1 of G. pyriformis (GpAMT1) and GpAMT2 were both plasma membrane localized, but only GpAMT1 transported ammonium. Neither protein transported the ammonium analogue methylammonium. Unexpectedly, GpAMT3 was localized in the vacuolar membrane, and it has as-yet-unknown transport characteristics. An unusual cysteine residue in the AMT signature of GpAMT2 and GpAMT3 was identified, and the corresponding residue was demonstrated to play an important role in ammonium transport. Surprisingly, each of the three AMTs of G. pyriformis had very distinct features. The localization of an AMT in the yeast vacuolar membrane is novel, as is the described amino acid residue that clearly influences ammonium transport. The AMT characteristics might reflect adaptations to the lifestyle of glomeromycotan fungi.


Effect of antiviral treatment on serum markers of liver fibrosis in HIV-hepatitis C virus-coinfected patients: the Fibrovic 2 Study - ANRS HC02.
Non-invasive liver fibrosis scores have been proposed as alternatives to liver biopsy (LB) in hepatitis C virus (HCV)-infected patients. Here, we aimed to assess the effect of antiviral treatment on non-invasive serological markers of liver fibrosis in HIV-HCV-coinfected patients. We included 114 HIV-HCV-coinfected patients with LBs performed before and 6 months after the end of treatment (week 72; W72). Fibrotest, the Forn's index, age-platelet ratio index, SHASTA, FIB-4, Hepa-score and Fibrometer scores were assessed. There were 29 (25%) patients who achieved sustained virological response (SVR). At baseline (BL), all non-invasive fibrosis scores except the Forn's index did not show significantly lower values in SVR patients. At W72, all non-invasive scores, except Hepascore, showed a significant decrease in SVR patients (P<0.01). There was a significant difference in fibrosis stages on LBs between BL and W72 in SVR and non-SVR patients. In HIV-HCV-coinfected patients, HCV clearance is associated with a significant reduction in non-invasive fibrosis serological markers, which most likely reflect the histological improvement associated with SVR. If confirmed, such results will reinforce the reliability of these markers in the follow-up after HCV treatment.


Patterns of nucleotide misincorporations during enzymatic amplification and direct large-scale sequencing of ancient DNA.
Whereas evolutionary inferences derived from present-day DNA sequences are by necessity indirect, ancient DNA sequences provide a direct view of past genetic variants. However, base lesions that accumulate in DNA over time may cause nucleotide misincorporations when ancient DNA sequences are replicated. By repeated amplifications of mitochondrial DNA sequences from a large number of ancient wolf remains, we show that C/G-to-T/A transitions are the predominant type of such misincorporations. Using a massively parallel sequencing method that allows large numbers of single DNA strands to be sequenced, we show that modifications of C, as well as to a lesser extent of G, residues cause such misincorporations. Experiments where oligonucleotides containing modified bases are used as templates in amplification reactions suggest that both of these types of misincorporations can be caused by deamination of the template bases. New DNA sequencing methods in conjunction with knowledge of misincorporation processes have now, in principle, opened the way for the determination of complete genomes from organisms that became extinct during and after the last glaciation.


A chemokine-dependent stromal induction mechanism for aberrant lymphocyte accumulation and compromised lymphatic return in rheumatoid arthritis.
According to the current model for tissue-specific homing, specificity is conferred by the selective recruitment of lymphocyte populations from peripheral blood, based on their expression of chemokine and adhesion receptors (endothelial selection). In this study, we provide evidence for an alternative stromal induction mechanism that operates in chronic inflammation. We show that the human rheumatoid synovial microenvironment directly induces functional inflammatory (CCR5 and CXCR3) and constitutive (CCR7 and CXCR4) chemokine receptors on infiltrating CD4(+) T cells. Expression of the corresponding inflammatory chemokine ligands (CCL5 and CXCL11) was confined to stromal areas in the synovium. However, expression of the constitutive ligands (CCL19 and CXCL12) was inappropriately high on both vascular and lymphatic endothelium, suggesting that the vascular to lymphatic chemokine gradient involved in lymphatic recirculation becomes subverted in the rheumatoid synovium. These results challenge the view that leukocyte trafficking is regulated solely by selective recruitment of pre-existing chemokine receptor-positive cells from peripheral blood, by providing an alternative explanation based on aberrant lymphocyte retention and compromised lymphatic return.


Loss of host cell plasma membrane integrity following cell traversal by Plasmodium sporozoites in the skin.
Plasmodium sporozoites are able to migrate through host cells by breaching their plasma membrane and gliding inside their cytoplasm. This migratory activity, called cell traversal (CT), was studied in vivo mainly using mutant sporozoites lacking the ability to wound host cells, and thus to perform CT. However, direct evidence of CT activity in host tissues by wild-type sporozoites remains scarce. Here, we describe a double-wounding assay to dynamically image CT activity in vivo and monitor cell membrane integrity over time. Based on the incorporation kinetics of a first live cell-impermeant dye, propidium iodide, we could determine whether traversed cells repair their wounded membranes or not. A second impermeant dye, SYTOX Green, was used to confirm the transient or the permanent loss of membrane integrity of traversed cells. This assay allowed, for the first time, the direct observation of sporozoites wounding and traversing host skin cells and showed that, while some traversed cells resealed their membrane, most became irreversibly permeable to these live cell-impermeant dyes. In combination with the study of CT-deficient sporozoites and the use of specific host cell markers, this intravital assay will provide the means to identify the nature of the cells traversed by sporozoites and will thus contribute to elucidating the role of CT by apicomplexan parasites in the vertebrate host.


Miniaturized thin film glutamate and glutamine biosensors.
Integrated thin film biosensors were developed for the simultaneous measurement of L-glutamine and L-glutamate in a mu-flow cell. Due to a novel glutaminase with an activity optimum in the neutral pH range, direct monitoring of glutamine in a mammalian cell culture medium could be performed. The glutamine bienzyme sensor was prepared by co-immobilization of glutaminase with glutamate oxidase within a photopatterned poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogel membrane. The sensor response was linear in the concentration range of 50 mumol to 10 mmol glutamine/l. Additionally, a glutamate biosensor was integrated on the sensor chip for difference measurement of possible glutamate interferences. The sensor-chip could be used for at least 300 measurements without any alteration in the performance of its sensors. A new sensor-chip with an integrated flow cell provided the possibility of simultaneous measurement of four different parameters at a cell volume of 1 microliter. In order to complete the microsystem, and in order to obtain a "lab on chip", a battery operated surface mounted device (SMD) potentiostat was developed.


Factors associated with cure after therapy of clinical mastitis caused by Staphylococcus aureus.
One hundred and fifty-nine cases of clinical Staphylococcus aureus mastitis were analyzed to detect factors associated with bacteriological cure after therapy. On 100 Dutch dairy farms, data were collected from four clinical trials with five intramammary treatment regimes designed to treat beta-lactamase-positive pathogens. Infected quarters were treated three times, with a 12-h interval between treatments. Treatment was extended for 2 d if results of the trial treatment were, according to the owner, not satisfactory. The overall bacteriological cure rate was 52%. The bacteriological cure rate of clinical beta-lactamase-negative S. aureus mastitis was significantly higher than that of clinical beta-lactamase-positive S. aureus mastitis. Bacteriological cure was also significantly higher if somatic cell count of the cow was low at the milk recording prior to the onset of the clinical mastitis. The bacteriological cure rate of clinical beta-lactamase-negative S. aureus mastitis was also significantly higher after an extended treatment compared with no extended treatment. The seriousness of the various clinical symptoms and the bacteriological cure rate of clinical S. aureus mastitis were not associated.


Measuring proteins in H2O with 2D-IR spectroscopy.
The amide I infrared band of proteins is highly sensitive to secondary structure, but studies under physiological conditions are prevented by strong, overlapping water absorptions, motivating the widespread use of deuterated solutions. H/D exchange raises fundamental questions regarding the impact of increased mass on protein dynamics, while deuteration is impractical for biomedical or commercial applications of protein IR spectroscopy. We show that 2D-IR spectroscopy can avoid this problem because the 2D-IR amide I signature of proteins dominates that of water even at sub-millimolar protein concentrations. Using equine blood serum as a test system, we investigate the significant implications of being able to measure the spectroscopy and dynamics of proteins in water, demonstrating relevance in areas ranging from fundamental science to the clinic. Measurements of vibrational relaxation dynamics of serum proteins reveals that deuteration slows down the rate of amide I vibrational relaxation by >10%, indicating a dynamic impact of isotopic exchange in some proteins. The unique link between protein secondary structure and 2D-IR amide I lineshape allows differentiation of signals due to albumin and globulin protein fractions in serum leading to measurements of the biomedically-important albumin to globulin ratio (AGR) with an accuracy of ±4% across a clinically-relevant range. Furthermore, we demonstrate that 2D-IR spectroscopy enables differentiation of the structurally similar globulin proteins IgG, IgA and IgM, opening up a straightforward spectroscopic approach to measuring levels of serum proteins that are currently only accessible via biomedical laboratory testing.


Lentiviral gene therapy vector with UCOE stably restores function in iPSC-derived neutrophils of a CDG patient.
A recent gamma-retroviral clinical Chronic Granulomatous Disease (CGD) gene therapy (GT) trial achieved proof-of-concept but was accompanied by activation of oncogenes and transgene silencing. The ubiquitous chromatin opening element (UCOE) comprises the sequences of two divergently oriented house-keeping gene promoters and is known to have anti-silencing properties. In a screen we identified two novel UCOE constructs that prevent adjacent promoter methylation in P19 cells. Experiments were continued with the shorter UCOE constructs in induced pluripotent stem cells (iPSC) derived from a p47phox-deficient CGD patient. The iPSC line was transduced with the lentiviral GT vectors expressing P47 under the constitutively active SFFV promoter with UCOE element (UCOE_SF) and without UCOE element (SF) adjacent to the SFFV promoter. The iPSC were expanded before propagation towards neutrophils. 20 days after transduction the UCOE_SF vector was protected from methylation in iPSC as previously shown in P19 cells, whereas the SF vector was heavily methylated in iPSC. The UCOE_SF vector maintained stable transgene expression in iPSC, macrophages and neutrophils, whereas the SF vector was strongly silenced. The UCOE_SF vector stably restored ROS production in neutrophils, whereas for the SF vector the count of ROS producing cells was marginal. To conclude, we have shown that the prevention of transgene silencing by UCOE is functionally and mechanistically preserved upon terminal neutrophil differentiation.


Efficacy comparison of oral rehydration solutions containing either 90 or 75 millimoles of sodium per liter.
In a randomized trial, 62 infants 2 to 35 months of age with dehydration due to acute watery diarrhea were allocated to one of two groups: group A received solution A (World Health Organization-recommended oral rehydration solution), which contained (mmol/L): Na+ 90, K+ 20, Cl- 80, citrate3- 10, and glucose 110; group B received solution B (Pedialyte RS; Abbott Laboratories, North Chicago), which contained (in mmol/L): Na+ 75, K+ 20, Cl- 65, citrate3- 10, and glucose 139. Oral therapy was given until clinical signs of hydration status were normal. During the 48-hour trial, the following laboratory data were collected: blood gases, serum electrolytes, glucose, urea, and creatinine values and sodium and potassium concentrations in stool and urine; serial weights and clinical signs were also reported. Six of the 62 infants, three in each group, required intravenous fluids because of high stool output. Results of clinical outcome and normalization of altered serum electrolyte values were similar in both groups. During the 48-hour trial, eight patients in group A and four in group B had mild, asymptomatic hypernatremia. Pedialyte RS was found to be a safe glucose/electrolyte solution for oral rehydration therapy.


Bushehr Elderly Health (BEH) Programme, phase I (cardiovascular system).
The main objective of the Bushehr Elderly Health Programme, in its first phase, is to investigate the prevalence of cardiovascular risk factors and their association with major adverse cardiovascular events. Between March 2013 and October 2014, a total of 3000 men and women aged ≥ 60 years, residing in Bushehr, Iran, participated in this prospective cohort study (participation rate=90.2%). Baseline data on risk factors, including demographic and socioeconomic status, smoking and medical history, were collected through a modified WHO MONICA questionnaire. Vital signs and anthropometric measures, including systolic and diastolic blood pressure, weight, height, and waist and hip circumference, were also measured. 12-lead electrocardiography and echocardiography were conducted on all participants, and total of 10 cc venous blood was taken, and sera was separated and stored at -80 °C for possible future use. Preliminary data analyses showed a noticeably higher prevalence of risk factors among older women compared to that in men. Risk factor assessments will be repeated every 5 years, and the participants will be followed during the study to measure the occurrence of major adverse cardiac events. Moreover, the second phase, which includes investigation of bone health and cognition in the elderly, was started in September 2015. Data are available at the Persian Gulf Biomedical Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran, for any collaboration.


Retreatability of root canals filled with Gutta percha and a novel bioceramic sealer: A scanning electron microscopy study.
This study aimed to evaluate the residual filling material and the reestablishment of working length (WL) and apical patency after retreatment of BioRoot™ RCS, versus TotalFill® BC Sealer, and AH26, used in a single cone obturation technique. Sixty single-rooted permanent canines were instrumented up to size 40/.04 (Sendoline, Täby, Sweden). The samples were filled with GP/AH26 (Group 1), GP/TotalFill BC Sealer (Group 2), and GP/BioRoot RCS (Group 3). Eight teeth were used in the positive and negative control groups. Canals were retreated using the Protaper Universal retreatment files (Dentsply Maillefer, Ballaigues, Switzerland) and further instrumented up to size 50/.04 (Sendoline, Täby, Sweden). The ability to reestablish WL and apical patency was recorded. Furthermore, the samples were observed under a scanning electron microscope, in order to evaluate the residual filling material. Data were analyzed statistically with the Fisher's exact and the Kruskal-Wallis tests (P < 0.05). There was no statistically significant difference in the WL and patency recovery among the groups. Neither the intragroup comparison among canal thirds nor the intergroup comparison in each canal third or in total canal area revealed any significant differences in the percentage of the remaining filling material after retreatment. Residual material debris was observed in all samples regardless of the sealer used. All the sealers were removed to a similar extent. The WL and patency were reestablished sufficiently in all groups.


Enzyme-Mediated Modification of Single-Domain Antibodies for Imaging Modalities with Different Characteristics.
Antibodies are currently the fastest-growing class of therapeutics. Although naked antibodies have proven valuable as pharmaceutical agents, they have some limitations, such as low tissue penetration and a long circulatory half-life. They have been conjugated to toxic payloads, PEGs, or radioisotopes to increase and optimize their therapeutic efficacy. Although nonspecific conjugation is suitable for most in vitro applications, it has become evident that site specifically modified antibodies may have advantages for in vivo applications. Herein we describe a novel approach in which the antibody fragment is tagged with two handles: one for the introduction of a fluorophore or (18)F isotope, and the second for further modification of the fragment with a PEG moiety or a second antibody fragment to tune its circulatory half-life or its avidity. Such constructs, which recognize Class II MHC products and CD11b, showed high avidity and specificity. They were used to image cancers and could detect small tumors.


Athletic groin pain: a systematic review of surgical diagnoses, investigations and treatment.
Athletic groin pain requiring surgery remains a diagnostic and therapeutic challenge. This systematic review aims to identify the most common causes of groin pain in athletes requiring surgery. Additionally, it aims to further characterise their susceptible athlete profiles, common physical examination and imaging techniques, and surgical procedures performed. This will enable the orthopaedic sports medicine clinician/surgeon to best treat these patients. The electronic databases MEDLINE, PubMed and EMBASE were searched from database inception to 13 August 2014 for studies in the English language that addressed athletic groin pain necessitating surgery. The search was updated on 4 August 2015 to find any articles published after the original search. The studies were systematically screened and data were abstracted in duplicate, with descriptive data presented. A total of 73 articles were included within our study, with data from 4655 patients abstracted. Overall, intra-articular and extra-articular causes of groin pain in athletes requiring surgery were equal. The top five causes for pain were: femoroacetabular impingement (FAI) (32%), athletic pubalgia (24%), adductor-related pathology (12%), inguinal pathology (10%) and labral pathology (5%), with 35% of this labral pathology specifically attributed to FAI. Given the complex anatomy, equal intra-articular and extra-articular contribution, and potential for overlap of clinical entities causing groin pain leading to surgery in athletes, further studies are required to ascertain the finer details regarding specific exam manoeuvres, imaging views and surgical outcomes to best treat this patient population.


The Real-World Effectiveness and Safety of Vedolizumab for Moderate-Severe Crohn's Disease: Results From the US VICTORY Consortium.
We assessed the real-world effectiveness and safety of vedolizumab (VDZ) in moderate-severe Crohn's disease (CD). Retrospective cohort study of seven medical centers, from May 2014 to December 2015. Adults with moderate-severe CD treated with VDZ, with follow-up after initiation of therapy, were included. Using the multivariable Cox proportional hazard analyses, we identified independent predictors of clinical remission or mucosal healing with VDZ. Rates of serious infection (requiring antibiotics, resulting in discontinuation of VDZ, hospitalization or death) and serious adverse events (discontinuation of VDZ, hospitalization or death) were described quantitatively. We included 212 patients with moderate-severe CD (median age 34 years; 40% male; 90% tumor necrosis factor (TNF)-antagonist exposed) with a median follow-up (IQR) of 39 weeks (25-53). Twelve-month cumulative rates of clinical remission, mucosal healing, and deep remission (clinical remission+mucosal healing) were 35%, 63%, and 26%, respectively. Individuals with prior TNF-antagonist exposure (hazard ratio (HR) 0.40; 95% confidence interval (CI): 0.20-0.81), smoking history (HR 0.47; 95% CI: 0.25-0.89), active perianal disease (HR 0.49; 95% CI: 0.27-0.88), and severe disease activity (HR 0.54; 95% CI: 0.31-0.95) were less likely to achieve clinical remission. Those with prior TNF-antagonist exposure (HR 0.29; 95% CI: 0.12-0.73), and severe disease activity (HR 0.54; 95% CI: 0.31-0.95) were less likely to achieve mucosal healing. During 160 patient years of follow-up (PYF) and 1,433 VDZ infusions, 5 patients developed infusion reactions (3.5 per 1,000 infusions), 21 developed serious infections (13 per 100 PYF), and 17 developed serious adverse events (10 per 100 PYF). A minority of adverse events required discontinuation of therapy (6 per 100 PYF). VDZ is a safe and effective treatment option for moderate-severe CD in routine practice. Clinical remission and deep remission (clinical remission and mucosal healing) can be achieved in 1/3 of individuals, and a minority of individuals require discontinuation of therapy due to adverse events.


Cope's rule and the dynamics of body mass evolution in North American fossil mammals.
Body mass estimates for 1534 North American fossil mammal species show that new species are on average 9.1% larger than older species in the same genera. This within-lineage effect is not a sampling bias. It persists throughout the Cenozoic, accounting for the gradual overall increase in average mass (Cope's rule). The effect is stronger for larger mammals, being near zero for small mammals. This variation partially explains the unwavering lower size limit and the gradually expanding mid-sized gap, but not the sudden large increase in the upper size limit, at the Cretaceous-Tertiary boundary.


[Differences in cognitive functioning of men and women with a diagnosis of depression].
Every year about 100 million people around the world, manifested symptoms of depression. The prevalence of depressive disorders concern for at least 15% of adult women and 10% of men. The aim of the study was to evaluate the effectiveness of selected cognitive processes in a group of men and women with recurrent depressive disorders. The survey involved 30 people (age: 23-62 years, mean age = 50.46 years, SD = 8.97): 16 women (53.33%; age: 23-59 years, mean age = 51.16 years, SD = 4.74) and 14 men (46.67%, respectively, age: 23-62 years, mean age = 51.41 years, SD = 12.07) hospitalized in Department of Adult Psychiatry, Medical University of Łódź. The study used psychological tests designed to assess the effectiveness of cognitive processes (learning curve Luria, a test plot away, Stroop test, verbal test fluencji by Łucki) and Hamilton depression rating scale (HDRS, consists of 21-items). Observed no statistically significant differences between treatment groups in the trials conducted. Women in the study group obtained better results compared to men in the learning curve Luria, Stroop test and verbal fluency test by Łucki. Men came out better in the test plot of the road. There were no differences between treatment groups in severity of depression measured HDRS. The results showed no statistically significant differences between men and women in the trials conducted evaluating the effectiveness of cognitive functioning. Women in the study group obtained better results compared to men in the trials evaluating the effectiveness of short-and long-term verbal memory, verbal memory and verbal fluidity. Men in the test group proved better than women in tests assessing visual-motor coordination and visual-spatial memory operational.


Effects of morphological patterning on endothelial cell migration.
The migration of vascular endothelial cells (ECs) plays an important role in vascular remodeling. Here we studied the effects of cell morphology on the migration of bovine aortic ECs by culturing cells on micropatterned strips of collagen matrix (60-, 30-, and 15-microm wide). The spreading areas of the cells on 15- and 30-microm wide strips were 30% lower than those on 60-microm wide strips and unpatterned collagen. The cells on 15-microm wide strips completely aligned in the direction of the strip, and had significantly lower shape index than those in all other groups. On strips of all widths, ECs tended to migrate in the direction of strips. ECs on 15-microm wide strips had highest speed, particularly in the direction of the strip. Vinculin staining showed that the leading edge of ECs on 15-microm wide strips had focal adhesions that were oriented with their lamellipodial protrusion and the direction of cell migration; this arrangement of the focal adhesions may promote EC migration. The present study provides direct evidence on the role of cell morphology in EC migration, and will help us to understand the mechanisms of EC migration during angiogenesis and wound healing.


TU-E-BRB-02: A Decision Support Tool for SBRT Planning Using a Searchable DVH Database.
Develop a decision support tool that aids dosimetrists, physicians, and physicists in assessing and improving plan quality through comparison to plans previously used in similar clinical situations. Software was developed to capture and store DVHs and other clinically relevant treatment plan characteristics in a database. In addition to the plan DVH, the database contains a total of 24 plan characteristics including fractionation, prescribed dose, treatment volume, prior surgery, tumor position, and smoking history. DVH and other plan data was captured from the treatment planning system via exported dicom RT files. Structures in the plan were automatically matched by name to a list of standard structures using a system of regular expressions. Additional fields were entered manually using a simple java interface. As a support tool, a plan under development can be quickly compared to similar plans in the database based on selected plan characteristics. A plot displaying the current and historical DVHs provides an easy visual comparison. Our interface also provides statistics for comparison for each dose/volume level such as average, minimum, maximum and standard deviation. DVHs from 111 lung SBRT plans treated from 2009-2011 were imported in accordance with an approved IRB protocol. As an example of data comparisons that can be easily performed to guide plan evaluation, we examined plans prescribing 5400cGy in 3 fractions and found that tumors >7.5cc (n=34) had an average PTV coverage of 94.2% (range: 73.5-95.0%), and tumors =7.5cc (n=35) had an average PTV coverage of 94.9% (range: 81.6-99.6%). A searchable DVH database was constructed to provide planners, physicists, and physicians with a straightforward means of comparing plans against historic distributions of DVHs. In the future, outcome data will be included in the database to strengthen its functionality as a decision support and research tool.


Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry.
Aberrant glycosylation is associated with most of the diseases. Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Brain sections from a C57BL/6 mouse were imaged using this technique at a resolution of 100 μm. Forty-two N-glycans were analyzed from the mouse brain section. The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. Furthermore, glioblastoma tumor xenografts in a NOD/SCID mouse were imaged. Several glycans with differential expression in tumor versus normal brain tissues were identified. The MSI technique allows for imaging of N-glycans directly from FFPE sections. This method can potentially identify tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures.


Environmental Enrichment Elicits a Transient Rise of Bioactive C-Type Natriuretic Peptide in Young but Not Aged Rats.
Beneficial molecular and neuroplastic changes have been demonstrated in response to environmental enrichment (EE) in laboratory animals across the lifespan. Here, we investigated whether these effects extend to C-type Natriuretic Peptide (CNP), a widely expressed neuropeptide with putative involvement in neuroprotection, neuroplasticity, anxiety, and learning and memory. We determined the CNP response in 36 young (8-9 months) and 36 aged (22-23 months) male PVGc hooded rats that were rehoused with new cage mates in either standard laboratory cages or EE for periods of 14 or 28 days. Tissues were rapidly excised from four brain regions associated with memory formation (dorsal hippocampus, retrosplenial cortex, medial prefrontal cortex, and mammillary bodies) plus the occipital cortex and hypothalamus, and immediately frozen. Radioimmunoassay was used to measure bioactive CNP and the amino-terminal fragment of proCNP, NTproCNP. Because CNP but not NTproCNP is rapidly degraded at source, NTproCNP reflects CNP production whereas the ratio NTproCNP:CNP is a biomarker of CNP's local degradation rate. EE increased CNP at 14 days in all brain regions in young, but not old rats; this effect in young rats was lost at 28 days in all regions of interest. NTproCNP:CNP ratio, but not NTproCNP, was reduced in all regions by EE at 14 days in young rats, but not in old rats, which suggests a period of reduced degradation or receptor mediated clearance, rather than increased production of CNP in these young EE rats. Aged rats tended to show reduced NTproCNP:CNP ratios but this did not occur in dorsal hippocampus or mammillary bodies. This is the first study demonstrating modulation of CNP protein concentrations, and the effect of age, in response to environmental stimulation. Furthermore, it is the first to show that changes in degradation rate in vivo may be an important component in determining CNP bioactivity in neural tissues.


In silico modeling and evaluation of Gordonia alkanivorans for biodesulfurization.
The genus Gordonia is well known for its catabolic diversity and ability to transform several compounds including the various recalcitrant polyaromatic sulfur heterocycles (PASHs) found in the fossil fuels. In fact, some strains offer the unique ability to desulfurize even benzothiophene (BT) and other thiophenic compounds, which most of the commonly studied rhodococci strains cannot. In this work, we present the first genome scale metabolic model for G. alkanivorans, a desulfurizing strain, to enable a holistic study of its metabolism and comparison with R. erythropolis. Our model consists of 881 unique metabolites and 922 reactions associated with 568 ORFs/genes and 544 unique enzymes. It successfully predicts the growth rates from experimental studies and quantitatively elucidates the pathways for the desulfurization of the commonly studied sulfur compounds, namely dibenzothiophene (DBT) and benzothiophene (BT). Using our model, we identify the minimal media for G. alkanivorans, and show the significant effect of carbon sources on desulfurization with ethanol as the best source. Our model shows that the sulfur-containing amino acids such as cysteine and methionine decrease desulfurization activity, and G. alkanivorans prefers BT over DBT as a sulfur source. It also suggests that this preference may be driven by the lower NADH requirements for BT metabolism rather than the higher affinity of the transport system for BT. Our in silico comparison of R. erythropolis and G. alkanivorans suggests the latter to be a better desulfurizing strain due to its versatility for both BT and DBT, higher desulfurization activity, and higher growth rate.


The role of l-arabinose metabolism for Escherichia coli O157:H7 in edible plants.
Arabinose is a major plant aldopentose in the form of arabinans complexed in cell wall polysaccharides or glycoproteins (AGP), but comparatively rare as a monosaccharide. l-arabinose is an important bacterial metabolite, accessed by pectolytic micro-organisms such as Pectobacterium atrosepticum via pectin and hemicellulose degrading enzymes. However, not all plant-associated microbes encode cell-wall-degrading enzymes, yet can metabolize l-arabinose, raising questions about their use of and access to the glycan in plants. Therefore, we examined l-arabinose metabolism in the food-borne pathogen Escherichia coli O157:H7 (isolate Sakai) during its colonization of plants. l-arabinose metabolism (araBA) and transport (araF) genes were activated at 18 °C in vitro by l-arabinose and expressed over prolonged periods in planta. Although deletion of araBAD did not impact the colonization ability of E. coli O157:H7 (Sakai) on spinach and lettuce plants (both associated with STEC outbreaks), araA was induced on exposure to spinach cell-wall polysaccharides. Furthermore, debranched and arabinan oligosaccharides induced ara metabolism gene expression in vitro, and stimulated modest proliferation, while immobilized pectin did not. Thus, E. coli O157:H7 (Sakai) can utilize pectin/AGP-derived l-arabinose as a metabolite. Furthermore, it differs fundamentally in ara gene organization, transport and regulation from the related pectinolytic species P. atrosepticum, reflective of distinct plant-associated lifestyles.


Toll-like receptor (TLR) 2 and TLR4 differentially regulate doxorubicin induced cardiomyopathy in mice.
Recent evidence indicates that toll-like receptor (TLR) 2 and 4 are involved in the pathogenesis of dilated cardiomyopathy (DCM), but the exact mechanisms of their actions have not been elucidated. We explored the therapeutic potential of blocking TLRs in mice with established cardiomyopathy. Cardiomyopathy was generated by a single intraperitoneal injection of doxorubicin (10 mg/kg). Two weeks later, the mice were treated with TLR2 or TLR4 neutralizing antibody. Blocking TLR2, but not TLR4, activity not only reduced mortality, but also attenuated doxorubicin-induced cardiac dysfunction by 20% and inhibited myocardial fibrosis. To determine the differential effects of blocking TLR2 and TLR4 in chronic cardiomyopathy, mice were injected with doxorubicin (3.5 mg/kg) once a week for 8 weeks, followed by treatment with TLR2 or TLR4 neutralizing antibody for 40 days. Blocking TLR2 activity blunted cardiac dysfunction by 13% and inhibited cardiac fibrosis, which was associated with a significant suppression of myocardial inflammation. The underlying mechanism involved interrupting the interaction of TLR2 with its endogenous ligands, resulting in attenuation of inflammation and fibrosis. In contrast, blocking TLR4 exacerbated cardiac dysfunction and fibrosis by amplifying inflammation and suppressing autophagy. Our studies demonstrate that TLR2 and TLR4 play distinct roles in the progression of doxorubicin-induced DCM. TLR4 activity is crucial for the resolution of inflammation and cardiac fibrosis, while blocking TLR2 activity has therapeutic potential for the treatment of DCM.


The inhibitory effect of natural microflora of food on growth of Listeria monocytogenes in enrichment broths.
The aims of this study were to (i) compare the inhibitory effects of the natural microflora of different foods on the growth of Listeria monocytogenes during enrichment in selective and non-selective broths; (ii) to isolate and identify components of the microflora of the most inhibitory food; and (iii) to determine which of these components was most inhibitory to growth of L. monocytogenes in co-culture studies. Growth of an antibiotic-resistant marker strain of L. monocytogenes was examined during enrichment of a range of different foods in Tryptone Soya Broth (TSB), Half Fraser Broth (HFB) and Oxoid Novel Enrichment (ONE) Broth. Inhibition of L. monocytogenes was greatest in the presence of minced beef, salami and soft cheese and least with prepared fresh salad and chicken pâté. For any particular food the numbers of L. monocytogenes present after 24h enrichment in different broths increased in the order: TSB, HFB and ONE Broth. Numbers of L. monocytogenes recovered after enrichment in TSB were inversely related to the initial aerobic plate count (APC) in the food but with only a moderate coefficient of determination (R(2)) of 0.51 implying that microbial numbers and the composition of the microflora both influenced the degree of inhibition of L. monocytogenes. In HFB and ONE Broth the relationship between APC and final L. monocytogenes counts was weaker. The microflora of TSB after 24h enrichment of minced beef consisted of lactic acid bacteria, Brochothrix thermosphacta, Pseudomonas spp., Enterobacteriaceae, and enterococci. In co-culture studies of L. monocytogenes with different components of the microflora in TSB, the lactic acid bacteria were the most inhibitory followed by the Enterobacteriaceae. The least inhibitory organisms were Pseudomonas sp., enterococci and B. thermosphacta. In HFB and ONE Broth the growth of Gram-negative organisms was inhibited but lactic acid bacteria still reached high numbers after 24h. A more detailed study of the growth of low numbers of L. monocytogenes during enrichment of minced beef in TSB revealed that growth of L. monocytogenes ceased at a cell concentration of about 10(2)cfu/ml when lactic acid bacteria entered stationary phase. However in ONE Broth growth of lactic acid bacteria was slower than in TSB with a longer lag time allowing L. monocytogenes to achieve much higher numbers before lactic acid bacteria reached stationary phase. This work has identified the relative inhibitory effects of different components of a natural food microflora and shown that the ability of low numbers of L. monocytogenes to achieve high cell concentrations is highly dependent on the extent to which enrichment media are able to inhibit or delay growth of the more effective competitors.


Urachal carcinoma: surgical and chemotherapeutic options.
The urachal ligament is an embryologic remnant connecting the dome of the bladder to the umbilicus via the ligamentum commune. Autopsy series suggest that in approximately a third of subjects, the urachal remnant may persist with tubular or cystic structures. However, tumors of this site are extremely rare. Patients usually present with hematuria and upon imaging, have evidence of a cystic or solid structure in the bladder dome or in the bladder midline. If a biopsy confirms adenocarcinoma, these tumors should be considered an urachal cancer until proven otherwise. Although there are no prospective clinical trials reported to date, large single-institution reports suggest surgical resection with a partial cystectomy and en bloc resection of the urachal ligament with umbilicus as the treatment of choice in the setting of localized disease. Although there is currently no definitive role for neoadjuvant or adjuvant chemotherapy in this tumor, risk factors predicting progression may allow for the selection of patients at higher relapse risk for prospective studies. Unfortunately, there are many patients who present with metastatic disease that currently is not likely to be curable. There is no standard chemotherapy regimen for these patients; however, there is new-found hope with a currently accruing clinical trial exploring a 5-fluorouracil-based chemotherapy combination in this patient population.


Wood smoke particles from different combustion phases induce similar pro-inflammatory effects in a co-culture of monocyte and pneumocyte cell lines.
Exposure to particulate matter (PM) has been linked to several adverse cardiopulmonary effects, probably via biological mechanisms involving inflammation. The pro-inflammatory potential of PM depends on the particles' physical and chemical characteristics, which again depend on the emitting source. Wood combustion is a major source of ambient air pollution in Northern countries during the winter season. The overall aim of this study was therefore to investigate cellular responses to wood smoke particles (WSPs) collected from different phases of the combustion cycle, and from combustion at different temperatures. WSPs from different phases of the combustion cycle induced very similar effects on pro-inflammatory mediator release, cytotoxicity and cell number, whereas WSPs from medium-temperature combustion were more cytotoxic than WSPs from high-temperature incomplete combustion. Furthermore, comparisons of effects induced by native WSPs with the corresponding organic extracts and washed particles revealed that the organic fraction was the most important determinant for the WSP-induced effects. However, the responses induced by the organic fraction could generally not be linked to the content of the measured polycyclic aromatic hydrocarbons (PAHs), suggesting that also other organic compounds were involved. The toxicity of WSPs seems to a large extent to be determined by stove type and combustion conditions, rather than the phase of the combustion cycle. Notably, this toxicity seems to strongly depend on the organic fraction, and it is probably associated with organic components other than the commonly measured unsubstituted PAHs.


Presynaptic and postsynaptic ion channel expression in vestibular nuclei neurons after unilateral vestibular deafferentation.
Vestibular compensation refers to the recovery of function occurring after unilateral vestibular deafferentation, but some patients remain uncompensated. Similarly, more than half of the operated chickens compensate three days after unilateral vestibular ganglionectomy (UVG), but the rest remain uncompensated. This review focuses on the studies performed on the principal cells of the chick tangential nucleus after UVG. The tangential nucleus is a major avian vestibular nucleus whose principal cells are all second-order, vestibular reflex projection neurons participating in the vestibuloocular and vestibulocollic reflexes controlling posture, balance, and eye movements. Using whole-cell patch-clamp approach in brain slice preparations, spontaneous spike firing, ionic conductances, and spontaneous excitatory postsynaptic currents (sEPSCs) are recorded in principal cells from controls and operated chickens three days after UVG. In compensated chickens, the proportion of spontaneous spike firing principal cells and their spike discharge rate are symmetric on the lesion and intact sides, with the rates increased over controls. However, in the uncompensated chickens, the spike discharge rate increases on the lesion side, but not on the intact side, where only silent principal cells are recorded. In all the experimental groups, including controls, silent principal cells are distinguished from spontaneous spiking cells by smaller persistent sodium conductances and higher activation thresholds for the fast sodium channel. In addition, silent principal cells on the intact side of uncompensated chickens have larger dendrotoxin-sensitive potassium conductances, with a higher ratio of immunolabeling for surface/cytoplasmic expression of a dendrotoxin-sensitive, potassium channel subunit, Kv1.1. Finally, in compensated chickens, sEPSC frequency is symmetric bilaterally, but in uncompensated chickens sEPSC frequency increased only on the lesion side, where the expression of Kv1.2 decreased in synaptotagmin-labeled terminal profiles on the principal cell bodies. Altogether, the specific sodium and potassium channels important for the development of spike firing pattern and/or presynaptic glutamate release on vestibular reflex projection neurons may be critically involved in changing postsynaptic neuron excitability after vestibular deafferentation.


Frictional adhesion: A new angle on gecko attachment.
Directional arrays of branched microscopic setae constitute a dry adhesive on the toes of pad-bearing geckos, nature's supreme climbers. Geckos are easily and rapidly able to detach their toes as they climb. There are two known mechanisms of detachment: (1) on the microscale, the seta detaches when the shaft reaches a critical angle with the substrate, and (2) on the macroscale, geckos hyperextend their toes, apparently peeling like tape. This raises the question of how geckos prevent detachment while inverted on the ceiling, where body weight should cause toes to peel and setal angles to increase. Geckos use opposing feet and toes while inverted, possibly to maintain shear forces that prevent detachment of setae or peeling of toes. If detachment occurs by macroscale peeling of toes, the peel angle should monotonically decrease with applied force. In contrast, if adhesive force is limited by microscale detachment of setae at a critical angle, the toe detachment angle should be independent of applied force. We tested the hypothesis that adhesion is increased by shear force in isolated setal arrays and live gecko toes. We also tested the corollary hypotheses that (1) adhesion in toes and arrays is limited as on the microscale by a critical angle, or (2) on the macroscale by adhesive strength as predicted for adhesive tapes. We found that adhesion depended directly on shear force, and was independent of detachment angle. Therefore we reject the hypothesis that gecko toes peel like tape. The linear relation between adhesion and shear force is consistent with a critical angle of release in live gecko toes and isolated setal arrays, and also with our prior observations of single setae. We introduced a new model, frictional adhesion, for gecko pad attachment and compared it to existing models of adhesive contacts. In an analysis of clinging stability of a gecko on an inclined plane each adhesive model predicted a different force control strategy. The frictional adhesion model provides an explanation for the very low detachment forces observed in climbing geckos that does not depend on toe peeling.


Using a Delphi process to determine optimal care for patients with pancreatic cancer.
Overall 5-year survival for pancreatic cancer is ∼5%. Optimizing the care that pancreatic cancer patients receive may be one way of improving outcomes. The objective of this study was to establish components of care which Australian health professionals believe important to optimally manage patients with pancreatic cancer. Using a Delphi process, a multidisciplinary panel of 250 health professionals were invited to provide a list of factors they considered important for optimal care of pancreatic cancer patients. They were then asked to score and then rescore (from one [no importance/disagree] to 10 [very important/agree]) the factors. The mean and coefficient of variation scores were calculated and categorized into three levels of importance. Overall, 63 (66% of those sent the final questionnaire; 25% of those initially invited) health professionals from nine disciplines completed the final scoring of 55 statements/factors encompassing themes of presentation/staging, surgery and biliary obstruction, multidisciplinary team details and oncology. Mean scores ranged from 3.7 to 9.7 with the highest related to communication and patient assessment. There was substantial intra- and interdisciplinary variation in views about MDT membership and roles. Overall, the opinions of Australian health professionals reflect international guideline recommended care; however, they identified a number of additional factors focusing on where patients should be treated, the importance of clear communication and the need for multidisciplinary care which were not included in current clinical practice guidelines. Differences in priorities between specialty groups were also identified.


Cationic P-S-X cages (X=Br, I).
The first condensed-phase preparation of ternary P-Ch-X cations (Ch=O-Te, X=F-I) is reported: [P5S3X2]+, [P5S2X2]+, and [P4S4X]+ (X=Br, I). [P5S3X2]+ is formed from the reaction of the Ag+/PX3 reagent with P4S3. The [P5S3X2]+ ions have a structure that is related to P4S5 by replacing P=S by P+--X and S in the four-membered ring by P(X). We provide evidence that the active ingredient of the Ag+/PX3 reagent is the (H2CCl2)Ag-X-PX2+ cation. The latter likely reacts with the HOMO of P4S3 in a concerted HOMO-LUMO addition to give the P5S3X2+ ion as the first species visible in situ in the low-temperature 31P NMR spectrum. The [P5S3X2]+ ions are metastable at -78 degrees C and disproportionate at slightly higher temperatures to give [P5S2X2]+ and [P4S4X]+, probably with the extrusion of 1/n (PX)n (X=Br, I). All six new cage compounds have been characterized by multinuclear NMR spectroscopy and, in part, by IR or Raman spectroscopy. The [P5S2X2]+ salts have a nortricyclane skeleton and were also characterized by X-ray crystallography. The structure of the [P4S4X]+ ion is related to that of P4S5 in that the exo-cage P=S bond is replaced by an isoelectronic P+--X moiety.