A Highly Effective Component Vaccine against Nontyphoidal Salmonella enterica Infections.
Nontyphoidal Salmonella enterica (NTS) infections are a major burden to global public health, as they lead to diseases ranging from gastroenteritis to systemic infections and there is currently no vaccine available. Here, we describe a highly effective component vaccine against S. enterica serovar Typhimurium in both gastroenteritis and systemic murine infection models. We devised an approach to generate supernatants of S. enterica serovar Typhimurium, an organism that is highly abundant in virulence factors. Immunization of mice with this supernatant resulted in dramatic protection against a challenge with serovar Typhimurium, showing increased survival in the systemic model and decreased intestinal pathology in the gastrointestinal model. Protection correlated with specific IgA and IgG levels in the serum and specific secretory IgA levels in the feces of immunized mice. Initial characterization of the protective antigens in the bacterial culture supernatants revealed a subset of antigens that exhibited remarkable stability, a highly desirable characteristic of an effective vaccine to be used under suboptimal environmental conditions in developing countries. We were able to purify a subset of the peptides present in the supernatants and show their potential for immunization of mice against serovar Typhimurium resulting in a decreased level of colonization. This component vaccine shows promise with regard to protecting against NTS, and further work should significantly help to establish vaccines against these prevalent infections. Salmonella enterica infections other than typhoid and paratyphoid fever are a major global health burden, as they cause high morbidity and mortality worldwide. Strategies that prevent Salmonella-related diseases are greatly needed, and there is a significant push for the development of vaccines against nontyphoidal Salmonella enterica serovars. In this work, we describe an S. Typhimurium supernatant-derived vaccine that is effective in reducing bacterial colonization in mouse models of gastroenteritis as well as invasive disease. This is a component vaccine that shows high stability to heat, a feature that is important for use under suboptimal conditions, such as those found in sub-Saharan Africa.


Physiological and biochemical basis of clinical liver function tests: a review.
To review the literature on the most clinically relevant and novel liver function tests used for the assessment of hepatic function before liver surgery. Postoperative liver failure is the major cause of mortality and morbidity after partial liver resection and develops as a result of insufficient remnant liver function. Therefore, accurate preoperative assessment of the future remnant liver function is mandatory in the selection of candidates for safe partial liver resection. A MEDLINE search was performed using the key words "liver function tests," "functional studies in the liver," "compromised liver," "physiological basis," and "mechanistic background," with and without Boolean operators. Passive liver function tests, including biochemical parameters and clinical grading systems, are not accurate enough in predicting outcome after liver surgery. Dynamic quantitative liver function tests, such as the indocyanine green test and galactose elimination capacity, are more accurate as they measure the elimination process of a substance that is cleared and/or metabolized almost exclusively by the liver. However, these tests only measure global liver function. Nuclear imaging techniques ((99m)Tc-galactosyl serum albumin scintigraphy and (99m)Tc-mebrofenin hepatobiliary scintigraphy) can measure both total and future remnant liver function and potentially identify patients at risk for postresectional liver failure. Because of the complexity of liver function, one single test does not represent overall liver function. In addition to computed tomography volumetry, quantitative liver function tests should be used to determine whether a safe resection can be performed. Presently, (99m)Tc-mebrofenin hepatobiliary scintigraphy seems to be the most valuable quantitative liver function test, as it can measure multiple aspects of liver function in, specifically, the future remnant liver.


Solid-state NMR of quadrupolar nuclei for investigations into supported organometallic catalysts: scope and frontiers.
The rational development of heterogeneous catalysts is a prerequisite for improving current catalytic technologies. One of the most successful approaches to achieve this goal is the grafting of organometallic complexes onto a given support, via surface organometallic chemistry (SOMC). In such cases, the immobilization of (transition) metal centres onto an inorganic support affords single-site catalysts with precise tuning of the coordination sphere and thus target-specific catalytic reactivity. In this context, the precise characterization of the surface sites by several spectroscopic techniques in parallel is of paramount importance. Solid-state NMR, in particular for half-integer quadrupolar nuclei, is a method that has increased its scope in the field of catalysis, especially due to recent methodological breakthroughs, occasionally combined with DFT calculations. In this tutorial review, we provide an overview of the most robust and efficient NMR experiments for half-integer quadrupolar nuclei, from simple one-dimensional acquisition under static and MAS conditions up to high-resolution and heteronuclear correlation two-dimensional spectra, as illustrated by selected examples.


Polymer swelling, drug mobilization and drug recrystallization in hydrating solid dispersion tablets studied by multinuclear NMR microimaging and spectroscopy.
Despite the advantages offered by solid dispersions, the marketed products based on this technology are few. The most frequent concern is the stability of the amorphous drug. The state of the drug in solid dispersions is, in general, poorly characterized as the number of characterization techniques available to monitor nanometer-sized drug particles embedded in a matrix are limited. Here we present a combination of localized NMR spectroscopic and NMR imaging techniques which allow in situ monitoring of the state of the drug during tablet disintegration and dissolution. (19)F NMR relaxation is shown to be sensitive to both the crystalline/amorphous state and the size of the model nanoparticles made of the drug substance flutamide. The time course of drug mobilization and recrystallization is detected with spatial resolution within swelling solid dispersion tablets. Comparing results from spatially resolved (19)F, (2)H and (1)H NMR experiments, recrystallization is related to its enabling factors such as local hydration level and local mobility of the polymer matrix. The initially amorphous drug may recrystallize either by nanoparticle coalescence or by ripening of crystalline grains.


Differences in Central Corneal Thickness between Spectral Domain-Optical Coherence Tomography and Ultrasound Pachymetry in Patients with Dry Eye Disease.
Purpose. To compare central corneal thickness (CCT) values via Spectral Domain-Optical Coherence Tomography (SD-OCT) and ultrasonic pachymetry in patients with severe dry eye disease (DED) to determine the level of agreement between these 2 methods. Methods. The paired samples t-test was used to compare CCT values in severe DED patients. Matching analysis between methods was performed using intraclass correlation coefficient (ICC). Intrasession reliability of the measurement methods was calculated via the concordance correlation coefficient (CCC), variation equivalent, and Pearson's correlation coefficient. The Bland-Altman procedure was used to graphically represent the differences between CCT values. Results. The study included 56 eyes of 24 female and 4 male patients. Mean age of the patients was 50.9 ± 11.3 years. Mean CCT via Cirrus SD-OCT was 523.82 ± 30.98 μm versus 530.050 ± 31.85 μm via ultrasonic pachymetry (paired samples t-test, P < 0.001). The Bland-Altman plot showed good agreement between the examiners. The ICC for repeatability was 0.974. The CCC between the 2 methods' CCT values was 0.973. The variation equivalent was 0.976 and Pearson's correlation coefficient was 99.3%, which also indicated high correlation between the 2 methods' measurements. Conclusions. The present findings show that in patients with severe DED Cirrus SD-OCT provides reliable intraobserver CCT values.


Backscattering of circularly polarized pulses.
Using numerical simulations of vector radiative transport, we examine time-resolved backscattering of circularly polarized plane waves normally incident upon a slab containing a random distribution of latex spheres in water. For large spheres the effect of polarization memory occurs a short time after first-order scattering and before depolarization. It is the result of successive near-forward-scattering events that maintain the incident wave's helicity. For moderately large scatterers it exhibits a simple dependence on the anisotropy factor. For larger spheres or those with higher refractive indices, it also depends on complicated angular and polarization characteristics of backscattering given by Mie theory.


The joint influence of competition and mutualism on the biodiversity of mutualistic ecosystems.
In the past years, there have been many advances -but also many debates- around mutualistic communities, whose structural features appear to facilitate mutually beneficial interactions and increase biodiversity, under some given population dynamics. However, most approaches neglect the structure of inter-species competition by adopting a mean-field perspective that does not deal with competitive interactions properly. Here, we build up a multilayer network that naturally accounts for mutualism and competition and show, through a dynamical population model and numerical simulations, that there is an intricate relation between competition and mutualism. Specifically, the multilayer structure is coupled to a dynamical model in which the intra-guild competitive terms are weighted by the abundance of shared mutualistic relations. We find that mutualism does not have the same consequences on the evolution of specialist and generalist species, and that there is a non-trivial profile of biodiversity in the parameter space of competition and mutualism. Our findings emphasize how the simultaneous consideration of positive and negative interactions derived from the real networks is key to understand the delicate trade-off between topology and biodiversity in ecosystems and call for the need to incorporate more realistic interaction patterns when modeling the structural and dynamical stability of mutualistic systems.


Evidence that unrejoined DNA double-strand breaks are not predominantly responsible for chromosomal radiosensitivity of AT fibroblasts.
To examine more fully the nature of chromosomal radiosensitivity in ataxia telangiectasia (AT) cells, we employed 24-color combinatorial painting to visualize 137Cs gamma-ray-induced chromosome-type aberrations in cells of two AT and one normal primary human fibroblast strains irradiated in log-phase growth. As a measure of misrejoined radiation-induced DSBs, we quantified exchange breakpoints associated with both simple and complex exchanges. As a measure of unrejoined DSBs, we quantified breakpoints from terminal deletions as well as deletions associated with incomplete exchange. For each of these end points, the frequency of damage per unit dose was markedly higher in AT cells compared to normal cells, although the proportion of total breaks that remained unrejoined was rather similar. The majority of breakpoints in both cell types were involved in exchanges. AT cells had a much higher frequency of complex exchanges compared to normal cells given the same dose, but for doses that resulted in approximately the same level of total breakpoints, the relative contribution from complex damage was also similar. We conclude that although terminal deletions and incomplete exchanges contribute to AT cell radiosensitivity, their relative abundance does not-in apparent contrast to the situation in lymphoblastoid cells-overwhelmingly account for the increased damage we observed in cycling AT fibroblasts. Thus, from a cytogenetic perspective, a higher level of unrepaired DSBs does not provide a universal explanation for the radiation-sensitive AT phenotype.


Evaluation of New Calibrated Pulse-Wave Analysis (VolumeViewTM/EV1000TM) for Cardiac Output Monitoring Undergoing Living Donor Liver Transplantation.
Intrapulmonary thermodilution technique using a pulmonary artery catheter is widely used for measuring cardiac output (CO) in patients undergoing liver transplantation. However, its invasiveness and associated complications have led to an interest in less invasive modalities. Thus, we aimed to evaluate whether the new calibrated pulse-wave analysis method monitoring (VolumeViewTM/EV1000TM) is interchangeable with intrapulmonary thermodilution technique. Twenty-eight patients undergoing living donor liver transplantation were enrolled in this prospective observational study. COs were recorded automatically by the two devices and compared simultaneously at 10-minute intervals. The agreement of absolute CO values and the tracking ability of CO changes trends were compared. A Bland-Altman analysis with percentage errors and concordance rate for trend analysis using both a 4-quadrant plot and a polar plot were performed on the data. A total of 375 paired datasets from 25 patients were included in analysis. COs measured by intrapulmonary thermodilution ranged from 3.8-13.7 L/min. The mean CO difference between the two techniques was 0.57 L/min, and the 95% limits of agreement were -0.98 L/min to 2.12 L/min with a percentage error of 42.3%. The percentage errors in the dissection, anhepatic, and reperfusion phase were 30.5%, 31.7%, and 27.4%, respectively. The concordance rate between the two techniques was 78.4%. The calibrated pulse-wave analysis and intrapulmonary thermodilution failed to show acceptable interchangeability in terms of both estimating CO and tracking CO changes during living donor liver transplantation.


T and Small Protrusion (TAP) vs Double-Kissing Crush Technique: Insights From In Vitro Models.
Percutaneous coronary interventions on complex bifurcation lesions may require implantation of two stents to appropriately treat diffuse side-branch (SB) disease. Comparisons among different bifurcation stenting techniques are continuously attempted by various study designs (bench tests, computer simulations, clinical studies). Among different techniques, double kissing crush (DKC) represents the last evolution for "crushing" while T and small Protrusion (TAP) represents the evolution of "T stenting". Both techniques are actually gaining popularity, but head-to-head comparisons are lacking. Two last generation drug-eluting stents (Synergy™, Boston Scientific, MA, USA and Ultimaster™,Terumo Corp., Japan) were implanted in left main bifurcation bench models using TAP (n = 6 sets) and DKC (n = 6 sets) techniques. A peristaltic pump with fresh porcine blood was used to perfuse the blood through the silicone model at a flow rate of 200 ml/min for 4 min. Optical coherence tomography (OCT) was used to assess stent struts geometry and thrombus formation. SB cross sectional area as well as SB obstruction did not significantly differ between the two techniques. Numerical (but not statistically significant) differences were found in terms of malapposed struts (fewer with TAP) and floating struts (fewer with DKC). Thrombus formation after blood perfusion was similar between TAP and DKC technique (1.53 ± 1.12 vs. 1.20 ± 1.01 mm2, p = 0.6). The result of the present in-vitro study shows the absence of significant difference between TAP and DKC in terms of stent struts apposition and acute thrombus formation potential. Despite the completely different technical steps required, both techniques have similar performance according to such articulated pre-clinical evaluation. WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT?: Due to its limited complexity, T and Protrusion (TAP) technique is considered the to-go technique for bifurcation lesions with good long-term results. Recently, double kissing crush (DKC) technique has been gaining popularity and demonstrated promising results in randomized clinical trials. WHAT DOES THIS STUDY ADD?: This in-vitro bench test study provides a unique detailed OCT comparison and local hemodynamic environment analysis of the two techniques. HOW MIGHT THIS IMPACT ON CLINICAL PRACTICE?: New insights of acute thrombogenicity and computational flow model simulation may guide percutaneous therapeutic strategies of bifurcation lesions.


Studies on the production of low density lipoproteins by perfused livers from nonhuman primates. Effect of dietary cholesterol.
Nonhuman primates consuming diets containing cholesterol develop coronary artery atherosclerosis that we have found to be highly correlated with an increase in the size and cholesteryl ester content of plasma low density lipoproteins (LDL). The present studies were designed to determine whether the enlarged plasma LDL are produced directly by the liver of cholesterol-fed monkeys. African green monkeys were fed a diet containing 40% of calories as butter fat and either 0.16 mg cholesterol/kcal (control diet) or 0.78 mg cholesterol/kcal (test diet). The livers of these monkeys were perfused by recirculation with a lipoprotein-free medium for 4 h. The rate of accumulation of perfusate cholesterol was linear and greater in liver perfusates from test diet-fed vs. control diet-fed monkeys and was positively correlated with both the plasma cholesterol concentration and LDL size in the donor animal. All perfusate d less than 1.063 g/ml lipoprotein subfractions from livers of test diet-fed monkeys were enriched in cholesteryl ester severalfold over the corresponding subfractions from control diet-fed monkeys and contained only the larger form of apolipoprotein B typical of plasma LDL. However, the perfusate lipoproteins in the LDL density range did not have an average size or composition typical of LDL from plasma. Rather, they were relatively enriched in phospholipid and unesterified cholesterol and were deficient in cholesteryl esters. In addition, perfusate high density lipoproteins were discoidal particles. These data show that the enzyme lecithin:cholesterol acyltransferase (LCAT) was essentially inactive in these perfusates and, as a result, the dietary cholesterol-induced enrichment of perfusate d less than 1.063 g/ml lipoproteins with cholesteryl esters probably resulted from increased hepatic secretion of cholesteryl esters and not from modification of lipoproteins by LCAT during recirculating perfusion. In spite of this increase, enlarged cholesteryl ester-rich LDL were not found in the perfusate, suggesting that large molecular weight plasma LDL are not directly secreted by the liver but instead probably result from further intravascular metabolism of cholesteryl ester-enriched hepatic precursor lipoproteins.


High-brightness diode laser arrays integrated with a phase shifter designed for single-lobe far-field pattern.
High-brightness, edge-emitting diode laser arrays integrated with a phase shifter have been designed and fabricated at a wavelength of about 910 nm. Stable out-of-phase mode is generated through coupling evanescently and converted to be nearly in-phase by the phase modulation from the phase shifter. With a very simple manufacture process, stable single-lobe far-field pattern is achieved in the slow axis when the continuous wave output power exceeds 460 mW/facet, and the divergence angle is only 2.7 times the diffraction-limited value. Such device shows a promising future for high-brightness application with low cost and easy fabrication.


Desensitization of endothelial P2Y1 receptors by PKC-dependent mechanisms in pressurized rat small mesenteric arteries.
Extracellular nucleotides play a crucial role in the regulation of vascular tone and blood flow. Stimulation of endothelial cell P2Y1 receptors evokes concentration-dependent full dilatation of resistance arteries. However, this GPCR can desensitize upon prolonged exposure to the agonist. Our aim was to determine the extent and nature of P2Y1 desensitization in isolated and pressurized rat small mesenteric arteries. The non-hydrolyzable selective P2Y1 agonist ADPbetaS (3 microM) was perfused through the lumen of arteries pressurized to 70 mmHg. Changes in arterial diameter and endothelial cell [Ca(2+)](i) were obtained in the presence and absence of inhibitors of protein kinase C (PKC). ADPbetaS evoked rapid dilatation to the maximum arterial diameter but faded over time to a much-reduced plateau closer to 35% dilatation. This appeared to be due to desensitization of the P2Y1 receptor, as subsequent endothelium-dependent dilatation to acetylcholine (1 microM) remained unaffected. Luminal treatment with the PKC inhibitors BIS-I (1 microM) or BIS-VIII (1 microM) tended to augment concentration-dependent dilatation to ADPbetaS (0.1-3 microM) and prevented desensitization. Another PKC inhibitor, Gö 6976 (1 microM), was less effective in preventing desensitization. Measurements of endothelial cell [Ca(2+)](i) in pressurized arteries confirmed the P2Y1 receptor but not M(3) muscarinic receptor desensitization. These data demonstrate for the first time the involvement of PKC in the desensitization of endothelial P2Y1 receptors in pressurized rat mesenteric arteries, which may have important implications in the control of blood flow by circulating nucleotides.


Efficient 1 kHz femtosecond optical parametric amplification in BiB(3)O(6) pumped at 800 nm.
We demonstrate efficient operation of a tunable femtosecond optical parametric amplifier based on BiB(3)O(6) pumped at 800 nm by a 1 kHz Ti:sapphire regenerative amplifier. The idler wavelength coverage extends to beyond 3 mum and the pulse duration at this wavelength is of the order of 110 fs. This new nonlinear borate crystal offers exceptionally high nonlinearity, making it a very promising candidate for power scaling of such frequency converters in the near-IR.


Anti-Pseudomonas aeruginosa activity of 1,10-phenanthroline-based drugs against both planktonic- and biofilm-growing cells.
The beneficial antimicrobial properties of 1,10-phenanthroline (phen)-based drugs, together with the imperative need to develop new chemotherapeutic options for prevention/treatment of infections caused by MDR Gram-negative bacteria, led us to evaluate the effects of phen, 1,10-phenanthroline-5,6-dione (phendione), [Ag(phendione)2]ClO4 and [Cu(phendione)3](ClO4)2·4H2O on planktonic- and biofilm-growing Pseudomonas aeruginosa. Thirty-two non-duplicated Brazilian clinical isolates of P. aeruginosa with distinct genetic backgrounds were used in all experiments. The effect of test compounds on planktonic bacterial proliferation was determined as recommended by CLSI protocol. The effect on biofilm formation was evaluated by crystal violet incorporation (biomass determination) and XTT (viability assay). Mature biofilm disorganization was evidenced by staining with crystal violet. Phen-based compounds presented anti-P. aeruginosa activity, but with different potencies concerning the geometric mean MIC: [Cu(phendione)3](2+) (7.76 μM) > [Ag(phendione)2](+) (14.05 μM) > phendione (31.15 μM) > phen (579.28 μM). MICs of each compound were similar irrespective of whether the P. aeruginosa isolates were susceptible or resistant to classical antimicrobials (ceftazidime, meropenem and imipenem). The pretreatment of bacteria with phen, phendione and phendione's metal derivatives at 0.5 × MIC value inhibited biofilm formation, particularly the use of [Cu(phendione)3](2+) and [Ag(phendione)2](+), which significantly reduced both biomass (48% and 44%, respectively) and viability (78% and 77%, respectively). The compounds studied also disrupted mature biofilm in a dose-dependent manner, especially [Ag(phendione)2](+) and [Cu(phendione)3](2+) (IC50, 9.39 and 10.16 μM, respectively). Coordination of phendione to Ag(+) and Cu(2+) represents a new promising group of anti-infective agents, which revealed a potent anti-P. aeruginosa action against both planktonic- and biofilm-growing cells.


Seventy-five years later, insulin remains Canada's major medical-research coup.
The world observes the 75th anniversary of the landmark discovery of insulin this year, a medical miracle that bears a large "made-in-Canada" stamp. CMAJ played a part in this story by publishing preliminary results of the first clinical trial on a human in March 1922. November is Diabetes Month, and to mark World Diabetes Day on Nov. 14 CMAJ once again heralds the accomplishment of the four discoverers of insulin.


Specialty Recertification.
The recognition of specialty boards started with the National Board of Medical Examiners and eventually gave rise to the Liaison Committee for Specialty Boards. The most appealing feature of any organization is its ability to provide quality of care. Because the timeframe for recertification may vary greatly among specialties, an approach that encourages physicians to participate in ongoing education between the 6- to 10-year certification deadlines is encouraged. Recertification demonstrates the physician's knowledge of new, innovative practices true competency, however, should encompass a physician's overall knowledge and ability to provide care that is both appropriate and effective. The standardization of healthcare is more evident now with healthcare reform underway, and with changes in the system. A physician's services need to be acceptable, and certification is a step in assuring that a standard of care is being met.


Speech and language impairment and oromotor dyspraxia due to deletion of 7q31 that involves FOXP2.
We report detailed clinical, cytogenetic, and molecular findings in a girl with a deletion of chromosome 7q31-q32. This child has a severe communication disorder with evidence of oromotor dyspraxia, dysmorphic features, and mild developmental delay. She is unable to cough, sneeze, or laugh spontaneously. Her deletion is on the paternally inherited chromosome and includes the FOXP2 gene, which has recently been associated with speech and language impairment and a similar form of oromotor dyspraxia in at least three other published cases. We hypothesize that our patient's communication disorder and oromotor deficiency are due to haploinsufficiency for FOXP2 and that her dysmorphism and developmental delay are a consequence of the absence of the other genes involved in the microdeletion. We propose that this patient, together with others reported in the literature, may define a new contiguous gene deletion syndrome encompassing the 7q31-FOXP2 region. Cytogenetic and molecular analysis of this region should be considered for other individuals displaying similar characteristics.


Sec1p binds to SNARE complexes and concentrates at sites of secretion.
Proteins of the Sec1 family have been shown to interact with target-membrane t-SNAREs that are homologous to the neuronal protein syntaxin. We demonstrate that yeast Sec1p coprecipitates not only the syntaxin homologue Ssop, but also the other two exocytic SNAREs (Sec9p and Sncp) in amounts and in proportions characteristic of SNARE complexes in yeast lysates. The interaction between Sec1p and Ssop is limited by the abundance of SNARE complexes present in sec mutants that are defective in either SNARE complex assembly or disassembly. Furthermore, the localization of green fluorescent protein (GFP)-tagged Sec1p coincides with sites of vesicle docking and fusion where SNARE complexes are believed to assemble and function. The proposal that SNARE complexes act as receptors for Sec1p is supported by the mislocalization of GFP-Sec1p in a mutant defective for SNARE complex assembly and by the robust localization of GFP-Sec1p in a mutant that fails to disassemble SNARE complexes. The results presented here place yeast Sec1p at the core of the exocytic fusion machinery, bound to SNARE complexes and localized to sites of secretion.


Upper gastrointestinal bleed associated with cholinesterase inhibitor use.
An 86-year-old man was admitted with a 3-day history of melaena and syncope. He was haemodynamically compromised and anaemic on presentation. His only medical history was mild Alzheimer's disease diagnosed 6 months prior. For this, he was on donepezil, a cholinesterase inhibitor (ChEI), with a recent dose increase 3 months earlier. After fluid resuscitation with packed red cells, an endoscopy was performed, which showed an acute duodenal ulcer. This was treated with a high-dose proton pump inhibitor. The patient recovered well and was discharged on donepezil with the addition of a gastro-protective proton pump inhibitor. In view of other absent risk factors of upper gastrointestinal haemorrhage, donepezil was the likely causative agent. ChEIs are associated with frequent side effects and increased hospitalisation due to central and peripheral increase in acetylcholine. With this case report, we review the literature of side effects related to ChEIs, where the mechanisms of action, complications and appropriate management are discussed.


Evidence for in vitro falsely-primed cDNAs that prevent specific detection of virus negative strand RNAs in dengue-infected cells: improvement by tagged RT-PCR.
The identification of cell types replicating dengue viruses is an important step towards the understanding of the pathophysiology of dengue severe forms. Since the detection of negative strand viral RNAs is the more reliable marker of active replication for single-strand positive sense RNA viruses, we reassessed the specificity of RT-PCR assays already developed to detect dengue negative strand RNAs. Studying mammalian Vero cells infected by a dengue-2 strain, it was shown that falsely-primed cDNAs are generated in vitro during the reverse transcription step and are amplified subsequently by PCR. Since this may compromise the specificity of existing RT-PCR systems, we developed a tagged RT-PCR assay and addressed the role of some critical factors in such a system. Optimization of the negative strand-specific tagged RT-PCR allowed to resolve the problems due to the PCR amplification of falsely-primed cDNAs. Using this assay it was possible to detect specifically negative strand RNAs as soon as 3h after Vero cells have been exposed to the dengue-2 strain and we showed that this system is highly specific. Thus, the present dengue negative strand-specific tagged RT-PCR assay may help to reassess viral replication in the context of dengue pathophysiology.


Repeated evolution of asymmetric genitalia and right-sided mating behavior in the Drosophila nannoptera species group.
Male genitals have repeatedly evolved left-right asymmetries, and the causes of such evolution remain unclear. The Drosophila nannoptera group contains four species, among which three exhibit left-right asymmetries of distinct genital organs. In the most studied species, Drosophila pachea, males display asymmetric genital lobes and they mate right-sided on top of the female. Copulation position of the other species is unknown. To assess whether the evolution of genital asymmetry could be linked to the evolution of one-sided mating, we examined phallus morphology and copulation position in D. pachea and closely related species. The phallus was found to be symmetric in all investigated species except D. pachea, which displays an asymmetric phallus with a right-sided gonopore, and D. acanthoptera, which harbors an asymmetrically bent phallus. In all examined species, males were found to position themselves symmetrically on top of the female, except in D. pachea and D. nannoptera, where males mated right-sided, in distinctive, species-specific positions. In addition, the copulation duration was found to be increased in the nannoptera group species compared to closely related outgroup species. Our study shows that gains, and possibly losses, of asymmetry in genital morphology and mating position have evolved repeatedly in the nannoptera group. Current data does not allow us to conclude whether genital asymmetry has evolved in response to changes in mating position, or vice versa.


Visual and auditory stimuli associated with swallowing activate mirror neurons: a magnetoencephalography study.
In the present study, we evaluated activated areas of the cerebral cortex with regard to the mirror neuron system during swallowing. To identify the activated areas, we used magnetoencephalography. Subjects were ten consenting volunteers. Swallowing-related stimuli comprised an animated image of the left profile of a person swallowing water with laryngeal elevation as a visual swallowing trigger stimulus and a swallowing sound as an auditory swallowing trigger stimulus. As control stimuli, a still frame image of the left profile without an additional trigger was shown, and an artificial sound as a false auditory trigger was provided. Triggers were presented at 3,000 ms after the start of image presentation. The stimuli were combined and presented and the areas activated were identified for each stimulus. With animation and still-frame stimuli, the visual association area (Brodmann area (BA) 18) was activated at the start of image presentation, while with the swallowing sound and artificial sound stimuli, the auditory areas BA 41 and BA 42 were activated at the time of trigger presentation. However, with animation stimuli (animation stimulus, animation + swallowing sound stimuli, and animation + artificial sound stimuli), activation in BA 6 and BA 40, corresponding to mirror neurons, was observed between 620 and 720 ms before the trigger. Besides, there were also significant differences in latency time and peak intensity between animation stimulus and animation + swallowing sound stimuli. Our results suggest that mirror neurons are activated by swallowing-related visual and auditory stimuli.


Synergistic antibacterial activity between Thymus vulgaris and Pimpinella anisum essential oils and methanol extracts.
Essential oils (EOs) and methanol extracts obtained from aerial parts of Thymus vulgaris and Pimpinella anisum seeds were evaluated for their single and combined antibacterial activities against nine Gram-positive and Gram-negative pathogenic bacteria: Staphylococcus aureus, Bacillus cereus, Escherichia coli, Proteus vulgaris, Proteus mirabilis, Salmonella typhi, Salmonella typhimurium, Klebsiella pneumoniae and Pseudomonas aeruginosa. The essential oils and methanol extracts revealed promising antibacterial activities against most pathogens using broth microdilution method. Maximum activity of Thymus vulgaris and Pimpinella anisum essential oils and methanol extracts (MIC 15.6 and 62.5mug/ml) were observed against Staphylococcus aureus, Bacillus cereus and Proteus vulgaris. Combinations of essential oils and methanol extracts showed an additive action against most tested pathogens especially Pseudomonas aeruginosa.


Provider-patient communication about prescription drug abuse: A qualitative analysis of the perspective of prescribers.
Background: Provider-patient communication underpins many initiatives aimed at reducing the public health burden associated with prescription drug abuse in the United States. The purpose of this qualitative analysis was to examine the characteristics of provider-patient communication about prescription drug abuse from the perspective of prescribers. Methods: From 2014 to 2015, 10 semi-structured interviews were conducted with a purposive sample of prescribers from multiple professions and medical fields in Central and South Central Appalachia. The interviews were conducted using a guide informed by Social Cognitive Theory and community theory research, audio-recorded, and transcribed verbatim. Thematic analysis, facilitated by NVivo 10 software, was used to generate themes. Results: Prescribers described 3 primary communication patterns with patients related to prescription drug abuse-informative, counteractive, and supportive. Prescribers also reported multiple factors-personal (e.g., education, experiences, and feelings of tension) and environmental (e.g., relationship with a patient, clinical resources, and policies on controlled prescription drugs)-that affect provider-patient communication and, by association, delivery of patient care related to prescription drug abuse. Conclusions: The findings suggest that provider-patient communication about prescription drug abuse is multidimensional and dynamic, characterized by multiple communication patterns and contributory factors. They have implications for (1) research aimed at advancing theoretical understanding of prescriber prescription drug abuse communication behaviors with patients and (2) interventions aimed at strengthening prescriber prescription drug abuse communication behaviors with patients.


Production of ectomycorrhizal mycelium peaks during canopy closure in Norway spruce forests.
*Here, species composition and biomass production of actively growing ectomycorrhizal (EM) mycelia were studied over the rotation period of managed Norway spruce (Picea abies) stands in south-western Sweden. *The EM mycelia were collected using ingrowth mesh bags incubated in the forest soil during one growing season. Fungal biomass was estimated by ergosterol analysis and the EM species were identified by 454 sequencing of internal transcribed spacer (ITS) amplicons. Nutrient availability and the fungal biomass in soil samples were also estimated. *Biomass production peaked in young stands (10-30 yr old) before the first thinning phase. Tylospora fibrillosa dominated the EM community, especially in these young stands, where it constituted 80% of the EM amplicons derived from the mesh bags. Species richness increased in older stands. *The establishment of EM mycelial networks in young Norway spruce stands requires large amounts of carbon, while much less is needed to sustain the EM community in older stands. The variation in EM biomass production over the rotation period has implications for carbon sequestration rates in forest soils.


Demonstration of methionine synthetase in intestinal mucosal cells of the rat.
Methionine synthetase was measured in the mucosal cells of the rat duodenum, jejunum and ileum by a previously employed method for mucosal cell isolation. No activity was found in these cells. When a dual buffer system for the isolation of villous and crypt cell population was substituted, however, methionine synthetase was found to be active in the duodenum, jejunum and ileum, both in the villous and crypt cell populations. The activity was significantly higher in the crypt cells than in the villous cells throughout the intestine, and higher levels were found in the ileum than in the duodenum or jejunum. As had been previously reported for the rat liver, nitrous oxide in vivo reduced the enzyme activity in both the villous and crypt cell populations, suggesting a role in vivo for the enzyme. We conclude that methionine synthetase is both present and active in the small intestinal mucosal cells of the rat.


A meta-analysis of responses of soil biota to global change.
Global environmental changes are expected to impact the abundance of plants and animals aboveground, but comparably little is known about the responses of belowground organisms. Using meta-analysis, we synthesized results from over 75 manipulative experiments in order to test for patterns in the effects of elevated CO(2), warming, and altered precipitation on the abundance of soil biota related to taxonomy, body size, feeding habits, ecosystem type, local climate, treatment magnitude and duration, and greenhouse CO(2) enrichment. We found that the positive effect size of elevated CO(2) on the abundance of soil biota diminished with time, whereas the negative effect size of warming and positive effect size of precipitation intensified with time. Trophic group, body size, and experimental approaches best explained the responses of soil biota to elevated CO(2), whereas local climate and ecosystem type best explained responses to warming and altered precipitation. The abundance of microflora and microfauna, and particularly detritivores, increased with elevated CO(2), indicative of microbial C limitation under ambient CO(2). However, the effects of CO(2) were smaller in field studies than in greenhouse studies and were not significant for higher trophic levels. Effects of warming did not depend on taxon or body size, but reduced abundances were more likely to occur at the colder and drier sites. Precipitation limited all taxa and trophic groups, particularly in forest ecosystems. Our meta-analysis suggests that the responses of soil biota to global change are predictable and unique for each global change factor.


Pharmacokinetics of orbifloxacin and its concentration in body fluids and in endometrial tissues of mares.
Pharmacokinetics and distribution of orbifloxacin into body fluids and endometrium was studied in 6 mares after intragastric (IG) administration at a single dose rate of 7.5 mg/kg body weight. Orbifloxacin concentrations were serially measured in serum, synovial fluid, peritoneal fluid, urine, cerebrospinal fluid, and endometrial tissues over 24 hours. Minimum inhibitory concentrations of orbifloxacin were determined for 120 equine pathogens over an 11-month period. The mean peak serum concentration (Cmax) was 2.41+/-0.30 microg/mL at 1.5 hours after administration and decreased to 0.17+/-0.01 microg/mL (Cmin) at 24 hours. The mean elimination half-life (t1/2) was 9.06+/-1.33 hours and area under the serum concentration vs time curve (AUC) was 20.54+/-1.70 mg h/L. Highest mean peritoneal fluid concentration was 2.15+/-0.49 microg/mL at 2 hours. Highest mean synovial fluid concentration was 1.17+/-0.28 microg/mL at 4 hours. Highest mean urine concentration was 536.67+/-244.79 microg/mL at 2 hours. Highest mean endometrial concentration was 0.72+/-0.23 microg/g at 1.5 hours. Mean CSF concentration was 0.46+/-0.55 microg/mL at 3 hours. The minimum inhibitory concentration of orbifloxacin required to inhibit 90% of isolates (MIC90) ranged from < or = 0.12 to > 8.0 microg/mL, with gram-negative organisms being more sensitive than gram-positive organisms. Orbifloxacin was uniformly absorbed in the 6 mares and was well distributed into body fluids and endometrial tissue. At a dosage of 7.5 mg/kg once a day, many gram-negative pathogens, such as Actinobacillus equuli, Escherichia coli, Pasteurella spp., and Salmonella spp. would be expected to be susceptible to orbifloxacin.


Identification and characterization of a mitogen-activated S6 kinase.
Treatment of Swiss mouse 3T3 cells with epidermal growth factor, orthovanadate, or serum results in the activation of a kinase that phosphorylates protein S6 of the 40S ribosomal subunit in vitro. This kinase is eluted as a single peak of activity from either a Mono Q anion-exchange column at 0.34 M NaCl or a Mono S cation-exchange column at 0.20 M NaCl. Treatment of the peak fraction from the Mono S column with phosphatase 2A completely abolishes the activity of the enzyme. The kinase appears to be distinct from protein kinase C, cAMP-dependent protein kinase, and two protease-activated kinases, PAK II and H4P. The kinase has been purified to apparent homogeneity and migrates as a single band at Mr 70,000 in NaDodSO4/polyacrylamide gels. The kinase exhibits the ability to autophosphorylate, and this activity directly parallels S6 phosphorylation activity on the final step of purification. In vitro, the kinase incorporates up to 5 mol of phosphate into S6, and the tryptic phosphopeptide maps obtained are equivalent to those from S6 phosphorylated in vivo. Most important, treatment of the purified kinase with phosphatase 2A results in complete inactivation of the enzyme, arguing that the activity of the kinase is directly controlled by phosphorylation.


CT abnormalities in schizophrenia. A preliminary study of their correlations with P300/P200 electrophysiological features and positive/negative symptoms.
Computed tomographic scans were scored blindly for the size of cerebrospinal fluid spaces in a group of nine medicated schizophrenics and a group of nine age-matched normal volunteers without psychiatric or medical problems. Overall, ten of the 18 computed tomography (CT) features measured were significantly enlarged in the schizophrenic group. These abnormal CT features were then correlated with electrophysiological and clinical measurements performed on the schizophrenic patients. Left sylvian fissure enlargement, thought to reflect temporal lobe tissue loss, was highly correlated with a left temporal scalp region feature of the auditory P300 measure (T3 electrode) that differentiated schizophrenics and normals, and both the left sylvian fissure enlargement and the P300 measure were highly correlated with positive symptoms (total score on the Scale for the Assessment of Positive Symptoms). Frontal superficial (cortical) sulcal enlargement was prominent in the schizophrenic group and was highly correlated with another electrophysiological measure, auditory P200, at left central scalp locations. There was no significant correlation between left sylvian fissure and frontal sulcal enlargement within the schizophrenic group, and intercorrelations between CT variables in the schizophrenic group were, in general, less significant than in the control group. Although we should be cautious about generalizability because of the small number of patients, these data are compatible with the hypothesis that different subgroups of schizophrenic pathological features are characterized by different CT, electrophysiological, and clinical presentations.


Reliable long-lasting depression interacts with variable short-term facilitation to determine corticostriatal paired-pulse plasticity in young rats.
Synaptic plasticity at corticostraital synapses is proposed to fine tune movment and improve motor skills. We found paired-pulse plasticity at corticostriatal synapses reflected variably expressed short-term facilitation blended with a consistent background of longer-lasting depression. Presynaptic modulation via neuotransmitter receptor activation was ruled out as a mechanism for long-lasting paired-pulse depression by examining the effect of selective receptor antagonists. EPSC amplitude and paired-pulse plasticity, however, was influenced by block of D2 dopamine receptors. Block of glutamate transport with l-transdicarboxylic acid (PDC) reduced EPSCs, possibly through a mechanism of AMPA receptor desensitization. Removal of AMPA receptor desensitization with cyclothiazide reduced the paired-pulse depression at long-duration interstimulus intervals (ISIs), indicating that AMPA receptor desensitization participates in corticostriatal paired-pulse plasticity. The low-affinity glutamate receptor antagonist cis-2,3-piperidine dicarboxylic acid (PDA) increased paired-pulse depression, suggesting that a presynaptic component also exists for long-lasting paired-pulse depression. Low Ca(2+)-high Mg(2+) or BAPTA-AM dramatically reduced the amplitude of corticostriatal EPSCs and both manipulations increased the expression of facilitation and, to a lesser extent, they reduced long-lasting paired-pulse depression. EGTA-AM produced a smaller reduction in EPSC amplitude and it did not alter paired-pulse facilitation, but in contrast to low Ca(2+) and BAPTA-AM, EGTA-AM increased long-lasting paired-pulse depression. These experiments suggest that facilitation and depression are sensitive to vesicle depletion, which is dependent upon changes in peak Ca(2+) (i.e. low Ca(2+)-high Mg(2+) or BAPTA-AM). In addition, the action of EGTA-AM suggests that basal Ca(2+) regulates the recovery from long-lasting paired-pulse depression, possibly thourgh a Ca(2+)-sensitive process of vesicle delivery.


Leaf and fruit essential oil compositions of Pimenta guatemalensis (Myrtaceae) from Costa Rica.
Pitnenta is a genus of flowering plants in the Myrtaceae family, which has about 15 species, mostly found in the Caribbean region of the Americas. Commonly used for culinary and medicinal purposes, the best known commercial species are allspice, P. dioica (P. officinalis) and bay rum, P. racemosa, but there is little information concerning P. guatemalensis. The aim of the present study was to identify the chemical composition of the leaf and fruit essential oils ofP. guatemalensis. The extraction of essential oils of P. guatemalensis growing wild in Costa Rica was carried out by the hydrodistillation method at atmospheric pressure, using a modified Clevenger type apparatus. The chemical composition of the oils was analyzed by capillary gas chromatographyflame ionization detector (GC/FID) and gas chromatography-mass spectrometry (GC/MS) using the retention indices on DB-5 type capillary column. A total of 103 and 63 compounds were identified in the leaf and fruit oils, respectively, corresponding to 96.8% and 86.1% of the total amount of the oils. The leaf oil consisted mainly of eugenol (72.8%), and mono- and sesquiterpene hydrocarbons (18.2%). Among terpenes the major components were beta-caryophyllene (8.2%) and terpinolene (3.0%). The fruit oil also consisted mainly of eugenol (74.7%) and minor amounts of oxygenated mono- and sesquiterpenes (7.3%), mainly caryophyllene oxide (3.3%). This is the first report of the chemical composition of the essential oils obtained from this plant species.


Fasting increases the extent of muscle necrosis in the mdx mouse.
1. The effects of fasting for 48 h were investigated in C57BL/10 (wild type) and age-matched C57BL/10 dystrophin-deficient (mdx) mice. 2. Fasting resulted in an increased percentage of necrotic fibres in muscles from the hindlimb and lumbar regions of mdx mice. The percentage of necrotic fibres of forelimb and chest muscles of mdx mice was unaltered by fasting. In wild-type mice, very few necrotic fibres were observed after fasting. 3. The necrotic changes in fasted mdx muscle were not accompanied by altered energy status as evaluated by muscle ATP and phosphocreatine concentrations. 4. A significantly decreased rectal temperature was observed in mdx but not in wild-type mice after fasting. 5. Fasting would normally be expected to cause a reduction in muscle fibre size. The high prevalence of necrosis in fasted mdx mice is therefore an unusual response that may be related to disturbance of the mechanisms which, in the fed state, compensate for the dystrophin deficiency in these animals.


Chemical- and Mechanical-Induced Lubrication Mechanisms during Hot Rolling of Titanium Alloys Using a Mixed Graphene-Incorporating Lubricant.
Hot rolling of titanium alloy currently is carried out without lubrication because of the surface defects. In order to explore an effective lubrication scheme to reduce friction and wear during hot rolling of titanium alloy, a mixed graphene-incorporating lubricant has been proposed to study its lubrication performance and mechanism. The tribological experiments were carried out by ball-disk friction and wear tester under hot-rolling parameters. Scanning electron microscopy (SEM), X-ray energy spectrum analyzer (EDS), X-ray powder diffractometer (XRD) and Raman analysis were used to analyse the surface and cross-section of the wear marks on the samples after the tribological experiments. The results show that the friction coefficient decreases up to about 35% compared with tests under dry and lubricated conditions. The surface quality of the wear marks is improved significantly after applying the proposed lubricant. The graphene which is embedded in the phosphate film can be effectively applied as a lubricating material to strengthen the lubricating film with less combustion loss at high temperatures. A chemical- and mechanical-induced lubrication mechanism for the hot rolling of titanium sheets has been proposed due to the synergistic lubrication effect of the graphene, ZrO2 nano particles and phosphate. It is of great significance and potential value to apply this proposed lubricant as an effective way to reduce the wear, friction and oxidation during the hot-rolling process of titanium alloy.


Assessment of the early risks of chorionic villus sampling.
A study was carried out to develop proficiency in performing chorionic villus sampling (CVS) and to determine whether the risk for miscarriage was so high as to preclude a randomized clinical trial comparing CVS with amniocentesis. A total of 202 women who had decided to have induced abortions volunteered to either undergo CVS (101 women) or be a control (101). There were no differences between the two groups in factors that may affect the rate of miscarriage. CVS was performed an average of 9.8 days before abortion. The rate of fetal loss was significantly higher in the CVS group (p = 0.009). An analysis of the results as a function of the physician's experience over time showed that there were distinct learning phases. It may take longer than is generally recognized to acquire the expertise necessary to perform CVS with the lowest risk possible. Caution should be exercised before diagnostic CVS is offered to women who plan to continue their pregnancies.


Long Noncoding RNA: Genomics and Relevance to Physiology.
The mammalian cell expresses thousands of long noncoding RNAs (lncRNAs) that are longer than 200 nucleotides but do not encode any protein. lncRNAs can change the expression of protein-coding genes through both cis and trans mechanisms, including imprinting and other types of transcriptional regulation, and posttranscriptional regulation including serving as molecular sponges. Deep sequencing, coupled with analysis of sequence characteristics, is the primary method used to identify lncRNAs. Physiological roles of specific lncRNAs can be examined using genetic targeting or knockdown with modified oligonucleotides. Identification of nucleic acids or proteins with which an lncRNA interacts is essential for understanding the molecular mechanism underlying its physiological role. lncRNAs have been reported to contribute to the regulation of physiological functions and disease development in several organ systems, including the cardiovascular, renal, muscular, endocrine, digestive, nervous, respiratory, and reproductive systems. The physiological role of the majority of lncRNAs, many of which are species and tissue specific, remains to be determined. © 2019 American Physiological Society. Compr Physiol 9:933-946, 2019.


Melioidosis in Far North Queensland. A clinical and epidemiological review of twenty cases.
During the last 4 years, 20 cases of clinical melioidosis were diagnosed in the geographical area between Tully and Thursday Island. Sixteen were diagnosed by culture of Pseudomonas pseudomallei, and four by positive serology with appropriate clinical features. Most cases occurred during or after a heavy wet season. All patients were adult, and males predominated. Farmers and stockmen represented predisposed populations due to their prolonged soil contact. Ten patients were white Australians, six were Aborigines and four were Torres Strait Islanders. Twelve cases were first diagnosed by positive blood culture and four by sputum culture. The primary site of infection was pulmonary in 14 cases, genitourinary tract in one case, subcutaneous tissues in one case, and joints in two cases. In cases of fulminating infection metastatic abscesses were commonly found in many organs; typically lungs, liver, kidneys and spleen. Six patients had acute fulminating disease and died. Fourteen patients successfully responded to appropriate therapy, but relapse occurred in three, all of whom had an alcohol problem and showed poor drug compliance. The presence of diabetes mellitus in six patients confirmed the important known association of these two diseases. In three fulminating and four subacute infections the serology was negative at the time of diagnosis by culture. Antibiotic therapy for the different forms of this disease is reviewed, and a laboratory protocol for the rapid reporting of positive culture results is included.


The incidence of infectious complications of central venous catheters at the subclavian, internal jugular, and femoral sites in an intensive care unit population.
The objective was to assess the risk of central venous catheter infection with respect to the site of insertion in an intensive care unit population. The subclavian, internal jugular, and femoral sites were studied. An epidemiologic, prospective, observational study. The setting is a well-functioning intensive care unit under a unified critical care medicine division in a university teaching hospital. Critical care medicine attendings and fellows covered on site 17 and 24 hrs per day, respectively. Patients were critically ill. All patients were triaged into the intensive care unit by on-site critical care medicine fellows. None. In an intensive care unit population, we prospectively studied the incidence of central venous catheter infection and colonization at the subclavian, internal jugular, and femoral sites. The optimal insertion site for each individual patient was selected by experienced intensive care physicians (critical care medicine attendings and fellows). All of the operators were proficient in inserting catheters at all three sites. Confounding factors were eliminated; there were a limited number of experienced operators inserting the catheters, a uniform protocol stressing strict sterile insertion was enforced, and standardized continuous catheter care was provided by dedicated intensive care nurses proficient in all aspects of central venous catheter care. Two groups of patients were analyzed. Group 1 was patients with one catheter at one site, and group 2 was patients with catheters at multiple sites. Group 1 was the primary analysis, whereas group 2 was supporting.A total of 831 central venous catheters and 4,735 catheter days in 657 patients were studied. The incidence of catheter infection (4.01/1,000 catheter days, 2.29% catheters) and colonization (5.07/1,000 catheter days, 2.89% catheters) was low overall. In group 1, the incidence of infection was subclavian: 0.881 infections/1,000 catheter days (0.45%), internal jugular: 0/1,000 (0%), and femoral: 2.98/1,000 (1.44%; p = .2635). The incidence of colonization was subclavian: 0.881 colonization/1,000 catheter days (0.45%), internal jugular: 2.00/1,000 (1.05%), and femoral: 5.96/1,000 (2.88%, p = .1338). There was no statistically significant difference in the incidence of infection and colonization or duration of catheters (p = .8907) among the insertion sites. In group 2, there was also no statistically significant difference in the incidence of infection and colonization among the three insertion sites. In an intensive care unit population, the incidence of central venous catheter infection and colonization is low overall and, clinically and statistically, is not different at all three sites when optimal insertion sites are selected, experienced operators insert the catheters, strict sterile technique is present, and trained intensive care unit nursing staff perform catheter care.


Donor-derived CD19-targeted T cells cause regression of malignancy persisting after allogeneic hematopoietic stem cell transplantation.
New treatments are needed for B-cell malignancies persisting after allogeneic hematopoietic stem cell transplantation (alloHSCT). We conducted a clinical trial of allogeneic T cells genetically modified to express a chimeric antigen receptor (CAR) targeting the B-cell antigen CD19. T cells for genetic modification were obtained from each patient's alloHSCT donor. All patients had malignancy that persisted after alloHSCT and standard donor lymphocyte infusions (DLIs). Patients did not receive chemotherapy prior to the CAR T-cell infusions and were not lymphocyte depleted at the time of the infusions. The 10 treated patients received a single infusion of allogeneic anti-CD19-CAR T cells. Three patients had regressions of their malignancies. One patient with chronic lymphocytic leukemia (CLL) obtained an ongoing complete remission after treatment with allogeneic anti-CD19-CAR T cells, another CLL patient had tumor lysis syndrome as his leukemia dramatically regressed, and a patient with mantle cell lymphoma obtained an ongoing partial remission. None of the 10 patients developed graft-versus-host disease (GVHD). Toxicities included transient hypotension and fever. We detected cells containing the anti-CD19-CAR gene in the blood of 8 of 10 patients. These results show for the first time that donor-derived allogeneic anti-CD19-CAR T cells can cause regression of B-cell malignancies resistant to standard DLIs without causing GVHD.


PIK3CA, BRAF, and PTEN status and benefit from cetuximab in the treatment of advanced colorectal cancer--results from NCIC CTG/AGITG CO.17.
Cetuximab improves survival in patients with K-ras wild-type advanced colorectal cancer. We examined the predictive and prognostic significance of additional biomarkers in this setting, in particular BRAF, PIK3CA, and PTEN. Available colorectal tumor samples were analyzed from the CO.17 study. BRAF mutations were identified in tumor-derived DNA by direct sequencing and PIK3CA mutations were identified using a high-resolution melting screen with confirmation by sequencing. PTEN expression by immunohistochemistry (IHC) was performed on tissue microarrays. For each biomarker, prognostic and predictive effects were examined using a Cox model with tests for treatment-biomarker interaction. A total of 572 patients with pretreated colorectal cancer were randomly assigned to receive cetuximab or best supportive care (BSC). Of 401 patients assessed for BRAF status, 13 (3.2%) had mutations. Of 407 patients assessed for PIK3CA status, 61 (15%) had mutations. Of 205 patients assessed for PTEN, 148 (72%) were negative for IHC expression. None of BRAF, PIK3CA, or PTEN was prognostic for overall or progression-free survival in the BSC arm. None was predictive of benefit from cetuximab, either in the whole study population or the K-ras wild-type subset. In the K-ras wild-type subgroup, the overall survival adjusted HR according to BRAF mutation status was 1.39 (interaction P = 0.69), PIK3CA mutation status HR = 0.79 (interaction P = 0.63), and PTEN expression HR = 0.75 (interaction P = 0.61). In chemotherapy-refractory colorectal cancer, neither PIK3CA mutation status nor PTEN expression were prognostic, nor were they predictive of benefit from cetuximab. Evaluation of predictive significance of BRAF mutations requires a larger sample size.


The economic burden of inpatient paediatric care in Kenya: household and provider costs for treatment of pneumonia, malaria and meningitis.
Knowledge of treatment cost is essential in assessing cost effectiveness in healthcare. Evidence of the potential impact of implementing available interventions against childhood illnesses in developing countries challenges us to define the costs of treating these diseases. The purpose of this study is to describe the total costs associated with treatment of pneumonia, malaria and meningitis in children less than five years in seven Kenyan hospitals. Patient resource use data were obtained from largely prospective evaluation of medical records and household expenditure during illness was collected from interviews with caretakers. The estimates for costs per bed day were based on published data. A sensitivity analysis was conducted using WHO-CHOICE values for costs per bed day. Treatment costs for 572 children (pneumonia = 205, malaria = 211, meningitis = 102 and mixed diagnoses = 54) and household expenditure for 390 households were analysed. From the provider perspective the mean cost per admission at the national hospital was US $95.58 for malaria, US $177.14 for pneumonia and US $284.64 for meningitis. In the public regional or district hospitals the mean cost per child treated ranged from US $47.19 to US $81.84 for malaria and US $54.06 to US $99.26 for pneumonia. The corresponding treatment costs in the mission hospitals were between US $43.23 to US $88.18 for malaria and US $ 43.36 to US $142.22 for pneumonia. Meningitis was treated for US $ 189.41 at the regional hospital and US $ 201.59 at one mission hospital. The total treatment cost estimates were sensitive to changes in the source of bed day costs. The median treatment related household payments within quintiles defined by total household expenditure differed by type of facility visited. Public hospitals recovered up to 40% of provider costs through user charges while mission facilities recovered 44% to 100% of costs. Treatments cost for inpatient malaria, pneumonia and meningitis vary by facility type, with mission and tertiary referral facilities being more expensive compared to primary referral. Households of sick children contribute significantly towards provider cost through payment of user fees. These findings could be used in cost effectiveness analysis of health interventions.


A cluster randomised controlled trial of an occupational therapy intervention for residents with stroke living in UK care homes (OTCH): study protocol.
The occupational therapy (OT) in care homes study (OTCH) aims to investigate the effect of a targeted course of individual OT (with task training, provision of adaptive equipment, minor environmental adaptations and staff education) for stroke survivors living in care homes, compared to usual care. A cluster randomised controlled trial of United Kingdom (UK) care homes (n = 90) with residents (n = 900) who have suffered a stroke or transient ischaemic attack (TIA), and who are not receiving end-of-life care. Homes will be stratified by centre and by type of care provided and randomised (50:50) using computer generated blocked randomisation within strata to receive either the OT intervention (3 months intervention from an occupational therapist) or control (usual care). Staff training on facilitating independence and mobility and the use of adaptive equipment, will be delivered to every home, with control homes receiving this after the 12 month follow-up.Allocation will be concealed from the independent assessors, but the treating therapists, and residents will not be masked to the intervention. Measurements are taken at baseline prior to randomisation and at 3, 6 and 12 months post randomisation. The primary outcome measure is independence in self-care activities of daily living (Barthel Activities of Daily Living Index). Secondary outcome measures are mobility (Rivermead Mobility Index), mood (Geriatric Depression Scale), preference based quality of life measured from EQ-5D and costs associated with each intervention group. Quality adjusted life years (QALYs) will be derived based on the EQ-5D scores. Cost effectiveness analysis will be estimated and measured by incremental cost effectiveness ratio. Adverse events will be recorded. This study will be the largest cluster randomised controlled trial of OT in care homes to date and will clarify the currently inconclusive literature on the efficacy of OT for stroke and TIA survivors residing in care homes. ISRCTN00757750.


A longitudinal study integrating population, care and social services data. The Swedish National study on Aging and Care (SNAC).
A large, national, long-term, longitudinal, multi-purpose study has been launched in Sweden--the Swedish National study on Aging and Care (SNAC). The study involves four research centers collecting data in four different areas of Sweden. The study consists of two parts: the population part and the care and services part. In the population part, a large, representative panel of elders in different age cohorts is followed over time to record and describe the aging process from different aspects. In the care and services part, a systematic, longitudinal, individually-based collection of data is performed concerning provision of care and services together with functional ability, specific health care problems, and living conditions of the recipients living in the area. The data collection in the population part of the SNAC is not yet completed. In the present article, some preliminary results are reported from the care and services part. These pertain to comparisons between the participating areas with respect to the prevalence of disability among those receiving care and social services in their ordinary homes and those receiving care in special accommodation. A comparison is also presented with regard to the amount of home help provided to subjects with a given disability. This project has several advantages. It is expected to generate a rich data base relevant for future research on aging and care and to have a direct impact on the future Swedish system of care and services for the elderly.


Sulforaphane inhibits extracellular, intracellular, and antibiotic-resistant strains of Helicobacter pylori and prevents benzo[a]pyrene-induced stomach tumors.
Gastric infection with Helicobacter pylori is a cosmopolitan problem, and is especially common in developing regions where there is also a high prevalence of gastric cancer. These infections are known to cause gastritis and peptic ulcers, and dramatically enhance the risk of gastric cancer. Eradication of this organism is an important medical goal that is complicated by the development of resistance to conventional antimicrobial agents and by the persistence of a low level reservoir of H. pylori within gastric epithelial cells. Moreover, economic and practical problems preclude widespread and intensive use of antibiotics in most developing regions. We have found that sulforaphane [(-)-1-isothiocyanato-(4R)-(methylsulfinyl)butane], an isothiocyanate abundant as its glucosinolate precursor in certain varieties of broccoli and broccoli sprouts, is a potent bacteriostatic agent against 3 reference strains and 45 clinical isolates of H. pylori [minimal inhibitory concentration (MIC) for 90% of the strains is <or=4 microg/ml], irrespective of their resistance to conventional antibiotics. Further, brief exposure to sulforaphane was bactericidal, and eliminated intracellular H. pylori from a human epithelial cell line (HEp-2). In complementary experiments, sulforaphane blocked benzo[a]pyrene-evoked forestomach tumors in ICR mice. This protection resulted from induction of phase 2 detoxication and antioxidant enzymes, and was abrogated in mice lacking the nrf2 gene, which regulates phase 2 enzymes. Thus, the dual actions of sulforaphane in inhibiting Helicobacter infections and blocking gastric tumor formation offer hope that these mechanisms might function synergistically to provide diet-based protection against gastric cancer in humans.


Testing for dependence on tree structures.
Tree structures, showing hierarchical relationships and the latent structures between samples, are ubiquitous in genomic and biomedical sciences. A common question in many studies is whether there is an association between a response variable measured on each sample and the latent group structure represented by some given tree. Currently, this is addressed on an ad hoc basis, usually requiring the user to decide on an appropriate number of clusters to prune out of the tree to be tested against the response variable. Here, we present a statistical method with statistical guarantees that tests for association between the response variable and a fixed tree structure across all levels of the tree hierarchy with high power while accounting for the overall false positive error rate. This enhances the robustness and reproducibility of such findings.


Potential for escape of live boll weevils (Coleoptera: Curculionidae) into cottonseed, motes, and cleaned lint at the cotton gin.
Reintroductions of the boll weevil, Anthonomus grandis grandis Boheman, into areas of the United States where it has been eradicated or suppressed are very expensive to mitigate. There is concern that a cotton gin in an eradication zone may serve as a site of boll weevil reintroductions when processing cotton harvested in a neighboring infested zone. Similarly, there is a question whether weevil-free areas can safely import gin products, such as cottonseed and baled lint, from infested areas without risking an introduction. Many countries require fumigation of imported U.S. cotton bales to protect against boll weevil introductions, costing the U.S. cotton industry millions of dollars annually. In previously reported experiments, we quantified the potential for boll weevils to survive passage through precleaning machinery in the gin. In this study, we quantified survival potential of boll weevils passing through the gin stand and segregating into the cottonseed, mote, or lint fractions. We also examined boll weevil survival when passed with ginned lint through a lint cleaner. We present a flow chart of experimentally determined survival potentials of boll weevils passing through the various subprocesses of the gin, from which one can calculate the risk of a live boll weevil reaching any point in the process. Our data show that there is virtually no chance of a boll weevil being segregated alive into the cottonseed or of one surviving in the lint to approach the bale press. Therefore, quarantine or fumigation of cottonseed and cotton bales to guard against boll weevil introductions is unnecessary.


Agreement among RTOG sarcoma radiation oncologists in contouring suspicious peritumoral edema for preoperative radiation therapy of soft tissue sarcoma of the extremity.
Peritumoral edema may harbor sarcoma cells. The extent of suspicious edema (SE) included in the treatment volume is subject to clinical judgment, balancing the risk of missing tumor cells with excess toxicity. Our goal was to determine variability in SE delineation by sarcoma radiation oncologists (RO). Twelve expert ROs were provided with T1 gadolinium and T2-weighted MR images of 10 patients with high-grade extremity soft-tissue sarcoma. Gross tumor volume, clinical target volume (CTV)3cm (3 cm longitudinal and 1.5 cm radial margin), and CTV2cm (2 cm longitudinal and 1 cm radial margin) were contoured by a single observer. Suspicious peritumoral edema, defined as abnormal signal on T2 images, was independently delineated by all 12 ROs. Contouring agreement was analyzed using the simultaneous truth and performance level estimation (STAPLE) algorithm and kappa statistics. The mean volumes of GTV, CTV2cm, and CTV3cm were, respectively, 130 cm(3) (7-413 cm(3)), 280 cm(3) and 360 cm(3). The mean consensus volume computed using the STAPLE algorithm at 95% confidence interval was 188 cm(3) (24-565 cm(3)) with a substantial overall agreement corrected for chance (mean kappa = 0.71; range: 0.32-0.87). The minimum, maximum, and mean volume of SE (excluding the GTV) were 4, 182, and 58 cm(3) (representing a median of 29% of the GTV volume). The median volume of SE not included in the CTV2cm and in the CTV3cm was 5 and 0.3 cm(3), respectively. There were 3 large tumors with >30 cm(3) of SE not included in the CTV3cm volume. Despite the fact that SE would empirically seem to be a more subjective volume, a substantial or near-perfect interobserver agreement was observed in SE delineation in most cases with high-grade soft-tissue sarcomas of the extremity. A median of 97% of the consensus SE is within the CTV2cm (99.8% within the CTV3cm). In a minority of cases, however, significant expansion of the CTVs is required to cover SE.


Mechanistic basis of immunotherapies for type 1 diabetes mellitus.
Type 1 diabetes (T1D) is an autoimmune disease for which there is no cure. The pancreatic beta cells are the source of insulin that keeps blood glucose normal. When susceptible individuals develop T1D, their beta cells are destroyed by autoimmune T lymphocytes and no longer produce insulin. T1D patients therefore depend on daily insulin injections for survival. Gene therapy in T1D aims at the induction of new islets to replace those that have been destroyed by autoimmunity. A major goal of T1D research is to restore functional beta cell mass while eliminating diabetogenic T cells in the hope of achieving insulin independence. Multiple therapeutic strategies for the generation of new beta cells have been under intense investigations. However, newly formed beta cells would be immediately destroyed by diabetogenic T cells. Therefore, successful islet induction therapy must be supported by potent immunotherapy that will protect the newly formed beta cells. Herein, we will summarize the current information on immunotherapies that aim at modifying T cell response to beta cells. We will first outline the immune mechanisms that underlie T1D development and progression and review the scientific background and rationale for specific modes of immunotherapy. Numerous clinical trials using antigen-specific strategies and immune-modifying drugs have been published, though most have proved too toxic or have failed to provide long-term beta cell protection. To develop an effective immunotherapy, there must be a continued effort on defining the molecular basis that underlies T cell response to pancreatic islet antigens in T1D.


Role for iron in reactive oxygen species-mediated cytotoxicity to cultured rat gastric mucosal cells.
The gastric epithelium is exposed to oxygen species that are generated within the lumen. Reactive oxygen species, enzymatically generated, cause injury to cultured rat gastric mucosal cells. Much interest has been focused on the role of iron in producing oxidant-mediated injury to the gastric mucosa, because iron is a catalyst that promotes the production of .OH possibly from O2-. and H2O2 (Haber-Weiss reaction) or from H2O2 alone (Fenton reaction). With the use of an iron chelator and an iron binding protein, we examined the role of iron in producing oxidant-mediated injury to cultured gastric mucosal cells. Reactive oxygen species and H2O2 were generated by hypoxanthine-xanthine oxidase and glucose-glucose oxidase, respectively, in buffer without iron. Pretreatment with deferoxamine diminished hypoxanthine-xanthine oxidase-induced 51Cr release from prelabeled cells, dose dependently. Furthermore, addition of deferoxamine to the reactive oxygen species-generating system also protected against the injury. However, apotransferrin (which binds extracellular iron) failed to protect cells. Pretreatment with .OH scavengers was partially protective. Depletion of glutathione with diethyl maleate enhanced reactive oxygen species-mediated cytolysis; such cytolysis was inhibited by deferoxamine. Deferoxamine also decreased 51Cr release induced by glucose-glucose oxidase. We conclude that intracellular iron plays a crucial role in mediating oxygen radical damage to gastric mucosal cells. The .OH, produced from H2O2 by the iron-catalyzed Fenton reaction, seems to be the main mediator of oxidant-induced cytotoxicity to gastric mucosal cells in vitro.


Repetitive transcranial magnetic stimulation (rTMS) in the treatment of obsessive-compulsive disorder (OCD) and Tourette's syndrome (TS).
There is evidence that motor and premotor cortex are hyperexcitable in obsessive-compulsive disorder (OCD) and Tourette's syndrome (TS). We tested whether low-frequency repetitive transcranial magnetic stimulation (rTMS) could normalize overactive motor cortical regions and thereby improve symptoms. Subjects with OCD or TS were treated with active rTMS to the supplementary motor area (SMA) for 10 daily sessions at 1 Hz, 100% of motor threshold, 1200 stimuli/day. Suggestions of clinical improvement were apparent as early as the first week of rTMS. At the second week of treatment, statistically significant reductions were seen in the YBOCS, YGTSS, CGI, HARS, HDRS, SAD, BDI, SCL-90, and SASS. Symptoms improvement was correlated with a significant increase of the right resting motor threshold and was stable at 3 months follow-up. Slow rTMS to SMA resulted in a significant clinical improvement and a normalization of the right hemisphere hyperexcitability, thereby restoring hemispheric symmetry in motor threshold.


Impact of chest CT on the clinical management of immunocompetent emergency department patients with chest radiographic findings of pneumonia.
The purpose of this study is to assess the impact on clinical decision making of chest computed tomography (CT) in immunocompetent emergency department (ED) patients with chest radiographic (CXR) findings of pneumonia. We retrospectively identified 1,373 patients from our ED who underwent chest CT between 7/05 and 6/06. Report of CXR within 24 h before CT were reviewed to identify patients with findings of pneumonia. The following were the exclusion criteria: recommendation of CT on CXR report and immunocompromised status on chart review. Fifty-one patients met the inclusion criteria: 26 women and 25 men, with a mean age of 60 (range 29-103) years. Age- and sex-matched controls from the ED with CXR findings of pneumonia who did not undergo CT were identified. Charts were reviewed for clinical presentation, management, and follow-up. Patient and control groups were compared using Fisher exact and paired Student's t tests. The patients were sicker than the controls with more signs and symptoms including auscultation abnormalities, 64 (33 of 51) vs 47% (24 of 51), abnormal sputum 32 (16 of 51) vs 0%, hypoxemia 22 (11 of 51) vs 2% (1 of 51), weight loss, 20 (10 of 51) vs 4% (2 of 51), and night sweats, 16 (8 of 51) vs 2% (1 of 51; p < 0.05 each). Clinical management, (based on CT findings in 31% [16 of 51]), was more extensive for patients than controls: antibiotics initiated 82 (41 of 51) vs 47% (24 of 51), antibiotics changed 29 (15 of 31) vs 0%, procedures performed 24 (12 of 51) vs 0%, and mean length of stay was 8 days vs less than 1 (p < 0.05, each). Sixteen percent (8 of 51) of the patients had alternative/additional diagnosis based on CT: pulmonary embolism, lung cancer, hypersensitivity pneumonitis, multiple myeloma, renal cell carcinoma, small bowel obstruction, lung nodule, and endobronchial mass (n = 1, each). Eight percent (4 of 51) of the patients and no controls were diagnosed with tuberculosis (p = 0.06). Immunocompetent ED patients with CXR findings of pneumonia who underwent chest CT were sicker than those who were not imaged with CT. Chest CT was often useful in guiding therapy or providing an alternative diagnosis.


The effects of exercise reminder software program on office workers' perceived pain level, work performance and quality of life.
In direct proportion to current technological developments, both the computer usage in the workplaces is increased and requirement of leaving the desk for an office worker in order to photocopy a document, send or receive an e-mail is decreased. Therefore, office workers stay in the same postures accompanied by long periods of keyboard usage. In recent years, with intent to reduce the incidence of work related musculoskeletal disorders several exercise reminder software programs have been developed. The purpose of this study is to evaluate the effectiveness of the exercise reminder software program on office workers' perceived pain level, work performance and quality of life. 39 healthy office workers accepted to attend the study. Participants were randomly split in to two groups, control group (n = 19) and intervention group (n = 20). Visual Analogue Scale to evaluate the perceived pain was administered all of the participants in the beginning and at the end of the study. The intervention group used the program for 10 weeks. Findings showed that the control group VAS scores remained the same, but the intervention group VAS scores decreased in a statistically significant way (p < 0.01). Results support that such exercise reminder software programs may help to reduce perceived pain among office workers. Further long term studies with more subjects are needed to describe the effects of these programs and the mechanism under these effects.


Facioscapulohumeral muscular dystrophy presenting with hypertrophic cardiomyopathy: a case study.
Only three facioscapulohumeral muscular dystrophy (FSHD) patients have been reported to have cardiomyopathy. An asymptomatic 38-year-old man was incidentally found to have electrocardiographic abnormalities. His echocardiogram demonstrated mild dilatation of the left ventricle and poor contractility. Cardiac histopathology indicated hypertrophic cardiomyopathy. Later he developed muscle weakness in the right arm. Scapular winging and asymmetrical facial weakness were evident. Muscle biopsy at the age of 44 years showed myopathic changes consistent with FSHD. His daughter had symptoms of infantile FSHD, which was genetically confirmed. This is the first report of an FSHD patient with biopsy-proven cardiomyopathy.


Using video feedback to improve early father-infant interaction: a pilot study.
Preventive interventions with parents of infants have tended to focus on mothers. Recent research focused on fathers suggests that their involvement in interventions might enhance effectiveness. One effective approach with mothers is the brief, home-based Video-feedback Intervention to promote Positive Parenting (VIPP). This paper is a report of a pilot study of VIPP with fathers to assess its feasibility. Five fathers were recruited from an existing longitudinal study of parents. The primary outcome was acceptability, assessed using a semi-structured questionnaire after completion of the intervention. All fathers completed all sessions of the intervention. Fathers rated the intervention as having had a significant impact on their understanding of their child's thoughts and feelings, and as having improved their communication and relationship with their baby. Fathers' feedback was generally positive. The flexibility to conduct sessions at home (or at fathers' places of work) and the flexible timing of sessions were identified as fundamental to successful delivery. The results of this pilot study are encouraging, as VIPP with fathers was feasible. In light of the modest sample size, and the use of a non-clinical sample, the intervention must be evaluated with larger, clinical samples to evaluate its efficacy with fathers.


Impact of New Chemotherapeutic agents on the outcome of Egyptian patients with Advanced Malignant Pleural Mesothelioma.
Patients with malignant pleural mesothelioma (MPM) are well known to have poor response to chemotherapy. Aim of this work was to evaluate the efficacy and safety of new chemotherapeutic agents for the treatment of Egyptian MPM patients. The first study was a non-randomized, open-label trial. It included 34 eligible patients who were assigned to receive either cisplatin/ pemetrexed or pemetrexed alone if cisplatin was contraindicated for a maximum of 8 cycles. In the second trial, 21 chemo-naïve patients with histologically proven advanced MPM were included. They received cisplatin and raltitrexed for a maximum of 6 cycles. In the first trial, the median age was 43.5 years (range 25-69), partial response (PR) was achieved in 37.5%, stable disease (SD) in 50%. Median time to progression (TTP) and overall survival (OS) were 7 and 14 months respectively. Survival at 1 year was 64.7%. No toxicity was observed in 17.6% of patients, grade 3-4 toxicity was evident in 11.8% (neutropenia), 8.8% (anemia), and 2.9% (vomiting and diarrhea). In the second trial, median age was 46 years (range 19- 71), PR was achieved in 23.2%, one complete remission (CR) was reported. SD was noticed in 61.9%. The median TTP and OS were 6 and 12 months respectively. Survival at 1 year was 51.6%. Both cisplatin/pemetrexed and cisplatin/ raltitrexed are effective and safe regimens in the treatment of MPM.


Accurate iron quantification in colloids and nanocomposites by a simple UV-Vis protocol.
The selection and comparative study is reported of calibration curves to quantify iron by a simple UV-Vis protocol based on the formation of iron (III) chloride complexes. The reliability of each calibration curve was evaluated using statistical and analytical parameters. The robustness of each calibration curve using superparamagnetic iron oxide nanoparticles (SPIONs) of different sizes and surface functionalization is demonstrated . We have also evaluated the effect of the particle coating and estimated the minimum time to ensure the full oxidation of iron (II) to (III) in sample solutions. Results from UV-Vis are comparable with those obtained from ICP-OES and from other spectroscopic techniques to quantify the iron. We advocate the proposed protocol as a simple and non-expensive route to determine accurately the iron content in colloidal and nanocomposite iron-based materials. Graphical abstract.


Salient motion features for video quality assessment.
Design of algorithms that are able to estimate video quality as perceived by human observers is of interest for a number of applications. Depending on the video content, the artifacts introduced by the coding process can be more or less pronounced and diversely affect the quality of videos, as estimated by humans. While it is well understood that motion affects both human attention and coding quality, this relationship has only recently started gaining attention among the research community, when video quality assessment (VQA) is concerned. In this paper, the effect of calculating several objective measure features, related to video coding artifacts, separately for salient motion and other regions of the frames of the sequence is examined. In addition, we propose a new scheme for quality assessment of coded video streams, which takes into account salient motion. Standardized procedure has been used to calculate the Mean Opinion Score (MOS), based on experiments conducted with a group of non-expert observers viewing standard definition (SD) sequences. MOS measurements were taken for nine different SD sequences, coded using MPEG-2 at five different bit-rates. Eighteen different published approaches related to measuring the amount of coding artifacts objectively on a single-frame basis were implemented. Additional features describing the intensity of salient motion in the frames, as well as the intensity of coding artifacts in the salient motion regions were proposed. Automatic feature selection was performed to determine the subset of features most correlated to video quality. The results show that salient-motion-related features enhance prediction and indicate that the presence of blocking effect artifacts and blurring in the salient regions and variance and intensity of temporal changes in non-salient regions influence the perceived video quality.


Oxidative damage in DNA bases revealed by UV resonant Raman spectroscopy.
We report on the use of the UV Raman technique to monitor the oxidative damage of deoxynucleotide triphosphates (dATP, dGTP, dCTP and dTTP) and DNA (plasmid vector) solutions. Nucleotide and DNA aqueous solutions were exposed to hydrogen peroxide (H2O2) and iron containing carbon nanotubes (CNTs) to produce Fenton's reaction and induce oxidative damage. UV Raman spectroscopy is shown to be maximally efficient to reveal changes in the nitrogenous bases during the oxidative mechanisms occurring on these molecules. The analysis of Raman spectra, supported by numerical computations, revealed that the Fenton's reaction causes an oxidation of the nitrogenous bases in dATP, dGTP and dCTP solutions leading to the production of 2-hydroxyadenine, 8-hydroxyguanine and 5-hydroxycytosine. No thymine change was revealed in the dTTP solution under the same conditions. Compared to single nucleotide solutions, plasmid DNA oxidation has resulted in more radical damage that causes the breaking of the adenine and guanine aromatic rings. Our study demonstrates the advantage of using UV Raman spectroscopy for rapidly monitoring the oxidation changes in DNA aqueous solutions that can be assigned to specific nitrogenous bases.


The value of cytological examination of the urine in the staging of invasive carcinoma of the cervix. A prospective study.
During the period July 1976 - December 1978 midstream urine specimens were collected from all new patients with invasive cervical carcinoma admitted to Tygerberg Hospital, Parowvallei, CP. All specimens were cytologically screened to try and establish any correlation between the cytological result an the clinical staging. A very high proportion (24.2%) of specimens unsuitable for screening and unacceptable false-negative and false-positive rates forced us to abandon this prospective study. From January 1979 to July 1980 the study was repeated on catheter specimens of urine. The final analysis of the second series revealed that a catheter specimen had definite advantages, but a detection rate of only 37.5% and a false-negative rate of 62.5% forced us to conclude that urinary cytological examination is at present too inaccurate to be of any value in the staging of invasive cervical carcinoma.


The microbiome regulates amygdala-dependent fear recall.
The amygdala is a key brain region that is critically involved in the processing and expression of anxiety and fear-related signals. In parallel, a growing number of preclinical and human studies have implicated the microbiome-gut-brain in regulating anxiety and stress-related responses. However, the role of the microbiome in fear-related behaviours is unclear. To this end we investigated the importance of the host microbiome on amygdala-dependent behavioural readouts using the cued fear conditioning paradigm. We also assessed changes in neuronal transcription and post-transcriptional regulation in the amygdala of naive and stimulated germ-free (GF) mice, using a genome-wide transcriptome profiling approach. Our results reveal that GF mice display reduced freezing during the cued memory retention test. Moreover, we demonstrate that under baseline conditions, GF mice display altered transcriptional profile with a marked increase in immediate-early genes (for example, Fos, Egr2, Fosb, Arc) as well as genes implicated in neural activity, synaptic transmission and nervous system development. We also found a predicted interaction between mRNA and specific microRNAs that are differentially regulated in GF mice. Interestingly, colonized GF mice (ex-GF) were behaviourally comparable to conventionally raised (CON) mice. Together, our data demonstrates a unique transcriptional response in GF animals, likely because of already elevated levels of immediate-early gene expression and the potentially underlying neuronal hyperactivity that in turn primes the amygdala for a different transcriptional response. Thus, we demonstrate for what is to our knowledge the first time that the presence of the host microbiome is crucial for the appropriate behavioural response during amygdala-dependent memory retention.


Pictorial Warning Labels and Memory for Cigarette Health-risk Information Over Time.
Pictorial cigarette warning labels are thought to increase risk knowledge, but experimental research has not examined longer-term effects on memory for health risks named in text. To investigate memory-consolidation predictions that high- versus low-emotion warnings would support better long-term memory for named cigarette health risks and to test a mediational model of warning-label effects through memory on risk perceptions and quit intentions. A combined sample of U.S.-representative adult smokers, U.S.-representative teen smokers/vulnerable smokers, and Appalachian-representative adult smokers were randomly assigned to a warning-label condition (High-emotion pictorial, Low-emotion pictorial, Text-only) in which they were exposed four times to nine warning labels and reported emotional reactions and elaboration. Memory of warning-label risk information, smoking risk perceptions, and quit intentions were assessed immediately after exposures or 6 weeks later. Recall of warning-label text was low across the samples and supported memory-consolidation predictions. Specifically, immediate recall was highest for Low-emotion warnings that elicited the least emotion, but recall also declined the most over time in this condition, leaving its 6-week recall lowest; 6-week recall was similar for High-emotion and Text-only warnings. Greater recall was associated with higher risk perceptions and greater quit intentions and mediated part of warning-label effects on these important smoking outcomes. High-emotion warnings had additional non-memory-related effects on risk perceptions and quit intentions that were superior to text-only warnings. High- but not Low-emotion pictorial warning labels may support the Food and Drug Administration's primary goal to "effectively convey the negative health consequences of smoking." NCT03375840.