Genome-Scale Phylogeny and Evolutionary Analysis of Ross River Virus Reveals Periodic Sweeps of Lineage Dominance in Western Australia, 1977-2014.
Ross River virus (RRV), an alphavirus of the Togaviridae family, is the most medically significant mosquito-borne virus of Australia. Past RRV phylogenetic and evolutionary analyses have been based on partial genome analyses only. Three geographically distinct RRV lineages, the Eastern, the Western, and the supposedly extinct North-Eastern lineage, were classified previously. We sought to expand on past phylogenies through robust genome-scale phylogeny to better understand RRV genetic diversity and evolutionary dynamics. We analyzed 106 RRV complete coding sequences, which included 13 genomes available on NCBI and 94 novel sequences derived for this study, sampled throughout Western Australia (1977-2014) and during the substantial Pacific Islands RRV epidemic (1979-1980). Our final data set comprised isolates sampled over 59 years (1959-2018) from a range of locations. Four distinct genotypes were defined, with the newly described genotype 4 (G4) found to be the contemporary lineage circulating in Western Australia. The prior geographical classification of RRV lineages was not supported by our findings, with evidence of geographical and temporal cocirculation of distinct genetic groups. Bayesian Markov chain Monte Carlo (MCMC) analysis revealed that RRV lineages diverged from a common ancestor approximately 94 years ago, with distinct lineages emerging roughly every 10 years over the past 50 years in periodic bursts of genetic diversity. Our study has enabled a more robust analysis of RRV evolutionary history and resolved greater genetic diversity that had been previously defined by partial E2 gene analysis.IMPORTANCE Ross River virus (RRV) causes the most common mosquito-borne infection in Australia and causes a significant burden of suffering to infected individuals as well as being a large burden to the Australian economy. The genetic diversity of RRV and its evolutionary history have so far only been studied using partial E2 gene analysis with a limited number of isolates. Robust whole-genome analysis has not yet been conducted. This study generated 94 novel near-whole-genome sequences to investigate the evolutionary history of RRV to better understand its genetic diversity through comprehensive whole-genome phylogeny. A better understanding of RRV genetic diversity will enable better diagnostics, surveillance, and potential future vaccine design.


Let the Diagram Speak: Compass Arcs and Visual Auxiliaries in Printed Diagrams of Euclid's Elements.
The printed Elements in the sixteenth century presented more practical and functional diagrams than those of previous manuscripts. Whereas conventional diagrams were limited to implementing the description of the text, the new diagrams introduced more concise constructions and visual auxiliaries. This change toward more practical and functional diagrams reflects the increased emphasis on the pedagogical value of the diagram. As is evident from the compass arcs upon the diagram, readers of the Elements were invited to draw their own diagrams, deviating from tradition and also from the text. Also, the increased visual auxiliaries such as correspondence markers, dotted lines, and stereoscopic presentations enabled the reader to read diagrams more easily. This backdrop of increased engagement with the diagram made it easier for mathematical novices (tyrones) to learn the Elements. These tool-based and auxiliaries-laden diagrams were more effective for teaching beginners than the earlier, less-functional diagrams. This paper explores the function of these new visual vocabularies and how they were circulated. This survey comprises a brief history of how diagrams began to have their say.


Gender-equitable attitudes, bystander behavior, and recent abuse perpetration against heterosexual dating partners of male high school athletes.
We assessed the relationship between gender attitudes, identified as a critical component of violence prevention, and abuse toward dating partners among adolescent male athletes. Our sample comprised 1699 athletes from 16 high schools in northern California who were surveyed between December 2009 and October 2010 in the larger Coaching Boys Into Men trial. We used logistic regression to assess the association between gender-equitable attitudes, bystander behavior, and recent abuse incidents. Athletes with more gender-equitable attitudes and greater intention to intervene were less likely (adjusted odds ratio [AOR] = 0.36; 95% confidence interval [CI] = 0.28, 0.46; and AOR = 0.60; 95% CI = 0.48, 0.75, respectively) and athletes who engaged in negative bystander behavior were more likely (AOR = 1.22, 95% CI = 1.10, 1.35) to perpetrate abuse against their female dating partners. Despite the shift among bystander intervention programs toward gender neutrality, our findings suggest a strong association between gender attitudes and dating violence. Programs designed for adolescents should include discussion of gender attitudes and target bystander behavior, because these components may operate on related but distinct pathways to reduce abuse.


Allelic and ectopic interactions in recombination-defective yeast strains.
Ectopic recombination in the yeast Saccharomyces cerevisiae has been investigated by examining the effects of mutations known to alter allelic recombination frequencies. A haploid yeast strain disomic for chromosome III was constructed in which allelic recombination can be monitored using leu2 heteroalleles on chromosome III and ectopic recombination can be monitored using ura3 heteroalleles on chromosomes V and II. This strain contains the spo13-1 mutation which permits haploid strains to successfully complete meiosis and which rescues many recombination-defective mutants from the associated meiotic lethality. Mutations in the genes RAD50, SPO11 and HOP1 were introduced individually into this disomic strain using transformation procedures. Mitotic and meiotic comparisons of each mutant strain with the wild-type parental strain has shown that the mutation in question has comparable effects on ectopic and allelic recombination. Similar results have been obtained using diploid strains constructed by mating MATa and MAT alpha haploid derivatives of each of the disomic strains. These data demonstrate that ectopic and allelic recombination are affected by the same gene products and suggest that the two types of recombination are mechanistically similar. In addition, the comparison of disomic and diploid strains indicates that the presence of a chromosome pairing partner during meiosis does not affect the frequency of ectopic recombination events involving nonhomologous chromosomes.


Experiencing affective music in eyes-closed and eyes-open states: an electroencephalography study.
In real life, listening to music may be associated with an eyes-closed or eyes-open state. The effect of eye state on listeners' reaction to music has attracted some attention, but its influence on brain activity has not been fully investigated. The present study aimed to evaluate the electroencephalographic (EEG) markers for the emotional valence of music in different eye states. Thirty participants listened to musical excerpts with different emotional content in the eyes-closed and eyes-open states. The results showed that participants rated the music as more pleasant or with more positive valence under an eyes-open state. In addition, we found that the alpha asymmetry indices calculated on the parietal and temporal sites reflected emotion valence in the eyes-closed and eyes-open states, respectively. The theta power in the frontal area significantly increased while listening to emotional-positive music compared to emotional-negative music under the eyes-closed condition. These effects of eye states on EEG markers are discussed in terms of brain mechanisms underlying attention and emotion.


Intraoperative spinal ultrasonography in two dogs with spinal disease.
Ultrasonography was performed during spinal surgery on two dogs that were suspected of having intramedullary lesions by myelography and magnetic resonance imaging. Ultrasonographically, the pathologic conditions of the spinal canal or cord were adequately imaged during surgery in both dogs. On the basis of ultrasonographic findings, a biopsy was obtained in Patient 1 and removal of the lesion was accomplished in Patient 2. Histopathologic diagnosis was myelomalacia in Patient 1 and spinal nephroblastoma in Patient 2. Intraoperative ultrasonography was demonstrated to be suitable for detecting intradural conditions, and, thus, is valuable for increasing the accuracy of biopsies or completeness of resections of intramedullary lesions.


Role of tumor necrosis factor-alpha in wound repair in human vocal fold fibroblasts.
Tumor necrosis factor-alpha (TNF-alpha) is an inflammatory cytokine and apoptotic molecule that appears to be a mediator in inflammation and fibrosis. The objective of this investigation was to examine the effects of TNF-alpha on 3D Carbylan-GSX in vitro cultured human vocal fold fibroblasts (hVFFs), to provide insight into the mechanism responsible for the improved vocal fold wound healing that has been previous reported with Carbylan-GSX treatment. In vitro cell culture. hVFF were cultured in 3D Carbylan-GSX and on polystyrene with different dosages of TNF-alpha (0, 0.1, 1, 10, and 100 ng/mL) with and without 10% fetal bovine serum (FBS). hVFF response to TNF-alpha was characterized by morphology, proliferation rates, and gene transcript levels for matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 2 (MMP2), tissue inhibitor of metalloproteinase 3 (TIMP3), collagen I, collagen III, fibronectin, and TNF-alpha receptor. In 3D Carbylan-GSX, TNF-alpha inhibited hVFF proliferation in a dose-dependent manner. TNF-alpha (0.1-100 ng/mL) was shown to significantly downregulate TIMP3 and extracellular matrix-related mRNA transcript levels for collagen III and fibronectin and to upregulate MMP1 and MMP2 expression, resulting in increased MMP/TIMP3 ratios. TNF-alpha receptor expression was significantly upregulated in Carbylan-GSX compared to control polystyrene. Responses were more marked in 10% FBS culture. After vocal fold injury, locally injected Carbylan-GSX can enhance the role of TNF-alpha in remodeling the lamina propria layer of the vocal fold, accelerating wound healing. Carbylan-GSX has potential as a new therapeutic approach that may lead to better treatment of vocal fold wound healing.


Phylogenetic diversity of bacteria in an earth-cave in Guizhou province, southwest of China.
The objective of this study was to analyze the phylogenetic composition of bacterial community in the soil of an earth-cave (Niu Cave) using a culture-independent molecular approach. 16S rRNA genes were amplified directly from soil DNA with universally conserved and Bacteria-specific rRNA gene primers and cloned. The clone library was screened by restriction fragment length polymorphism (RFLP), and representative rRNA gene sequences were determined. A total of 115 bacterial sequence types were found in 190 analyzed clones. Phylogenetic sequence analyses revealed novel 16S rRNA gene sequence types and a high diversity of putative bacterial community. Members of these bacteria included Proteobacteria (42.6%), Acidobacteria (18.6%), Planctomycetes (9.0%), Chloroflexi (Green nonsulfur bacteria, 7.5%), Bacteroidetes (2.1%), Gemmatimonadetes (2.7%), Nitrospirae (8.0%), Actinobacteria (High G+C Gram-positive bacteria, 6.4%) and candidate divisions (including the OP3, GN08, and SBR1093, 3.2%). Thirty-five clones were affiliated with bacteria that were related to nitrogen, sulfur, iron or manganese cycles. The comparison of the present data with the data obtained previously from caves based on 16S rRNA gene analysis revealed similarities in the bacterial community components, especially in the high abundance of Proteobacteria and Acidobacteria. Furthermore, this study provided the novel evidence for presence of Gemmatimonadetes, Nitrosomonadales, Oceanospirillales, and Rubrobacterales in a karstic hypogean environment.


Gaps of free-space optics beams with the Beer-Lambert law.
Lasers used in free-space optics propagate a beam within a truncated cone. Because of this shape, the intensity cannot follow the Beer-Lambert law. In the case of a homogeneous atmosphere, we calculate the gap from the cylinder case. We will see that the gap exists but is generally very weak and, therefore, that the use of the Beer-Lambert law is a justified approximation.


Loss of heterozygosity in 11q13-14 regions in gastric neuroendocrine tumors not associated with multiple endocrine neoplasia type 1 syndrome.
Loss of heterozygosity (LOH) at the MEN1 gene locus at 11q13 is commonly found in type II gastric carcinoid tumors, which are associated with multiple endocrine neoplasia type 1 (MEN-1). In contrast, information is scanty or absent for other types of gastric neuroendocrine tumors, represented by type I carcinoids (associated with chronic atrophic gastritis), type III (sporadic) carcinoids, and neuroendocrine carcinomas. Moreover, LOH analysis of the allelic region distal to the MEN1 gene, which is postulated to contain an additional tumor suppressor gene effective in MEN-1-associated and sporadic endocrine tumors, has never been performed. To clarify these issues, DNA extracted from archival tissue from 25 type I carcinoids, 4 type III carcinoids, and 2 neuroendocrine carcinomas was amplified by PCR, using primers for six polymorphic markers located on chromosome 11q13 (PYGM, D11S4946, and D11S913) and 11q14 (D11S916, D11S901, and D11S1365), for analysis of LOH. Allelic losses in the 11q13-14 region with at least two polymorphic markers were found in 12 of 25 (48%) type I carcinoids. When LOH was found in the 11q13 region, it was large and continuous and extended to the most telomeric marker investigated. In one tumor, retention of heterozygosity for markers in the MEN1 region and LOH for distal markers were observed. No LOH was found in three of four type III carcinoids. Large deletions in both the 11q13 and 11q14 regions were observed in both neuroendocrine carcinomas investigated. In conclusion, LOH in the 11q13-14 regions is frequently found in type I carcinoids and neuroendocrine carcinomas of the stomach, suggesting the involvement of the MEN1 gene and/or a more telomeric tumor suppressor gene in the pathogenesis of these non-MEN-1-associated neuroendocrine tumors. The low rate of LOH at 11q13-14 suggests the predominance of different genetic mechanisms in type III carcinoids, which also differ from other types of gastric carcinoids in the lack of a promoter role for gastrin.


Antifungals susceptibility pattern of Candida spp. isolated from female genital tract at the Yaoundé Bethesda Hospital in Cameroon.
Vaginal candidiasis is considered as an important public health problem worldwide and its incidence has increased nowadays. In recent years, inappropriate and disproportionate use of antifungal drugs, automedication as well as non compliance have caused drug resistance. This study aimed at determining the in vitro antifungal susceptibility patterns of Candida speciesisolated from female genital tract at Yaoundé Bethesda Hospital in Cameroon. Two hundred and fourthy five women (age range: 15 years to 49 years) attending the hospital were recruited between January and June 2014 in this cross sectional study. Vaginal smears were collected using sterile swabs from each participant and cultured on sabouraud dextrose agar supplemented with chloramphenico l 0.5%; identification of Candida spp. was performed following standard methods. The disk diffusion method was used for antifungal susceptibility testing. Out of the 245 vaginal smears collected, 94 (38.4%) strains of yeast were isolates among which 43 (45.7%) were Candida albicans and 51 (54.3%) were non albicans. The highest susceptibility of the isolates was seen for nystatin 62 (83.78%), ketoconazole 61 (82.43%) and fluconazole 60 (81.08%). Despite the noticeable resistance of Candida spp. isolates to miconazole and itraconazole, the results indicate that nystatin, ketoconazole and fluconazole are the drugs of choice for the therapy of vaginal candidiasis in this region.


Psychiatric symptoms associated with brief detention of adult asylum seekers in Canada.
To examine the association between brief detention and psychiatric symptom levels among adult asylum seekers. The Harvard Trauma Questionnaire and the Hopkins Symptoms Checklist-25 were used to assess psychiatric symptoms and premigration trauma exposure in 122 detained and 66 nondetained adult asylum seekers in Montreal and Toronto. After a mean detention of 31 days, the proportion of asylum seekers scoring above clinical cutpoints was significantly higher in the detained than the nondetained group for posttraumatic stress (χ² = 4.117, df = 1, P = 0.04), depression (χ² = 13.813, df = 1, P < 0.001), and anxiety (χ² = 4.567, df = 1, P = 0.03) symptoms. Hierarchical multiple regression models showed that posttraumatic stress, depression, and anxiety symptom levels were significantly higher among detained asylum seekers than among the nondetained comparison group, taking into account previous trauma and demographics. Incremental F was significant for the addition of detention status for all 3 models, indicating that detention contributed to increased symptom levels. For asylum seekers, even brief detention is associated with increased psychiatric symptoms. Governments should consider the many viable alternatives to incarceration of asylum seekers, such as temporary placement in a supervised residential facility, to minimize the risks of psychological harm to this vulnerable population.


The mouse cortical meninges are the site of immune responses to many different pathogens, and are accessible to intravital imaging.
A wide range of viral and microbial infections are known to cause meningitis, and there is evidence that the meninges are the gateway to pathogenic invasion of the brain parenchyma. Hence observation of these regions has wide application to understanding host-pathogen interactions. Interactions between pathogens and cells of the immune response can be modified by changes in their environment, such as suppression of the flow of blood and lymph, and, particularly in the case of the meninges, with their unsupported membranes, invasive dissection can alter the tissue architecture. For these reasons, intravital imaging through the unperforated skull is the method of choice. We give a protocol for a simple method of two-photon microscopy through the thinned cortical skull of the anesthetized mouse to enable real-time imaging with sub-micron resolution through the meninges and into the superficial brain parenchyma. In reporter mice in which selected cell types express fluorescent proteins, imaging after infection with fluorescent pathogens (lymphocytic choriomeningitis virus, Trypanosoma brucei or Plasmodium berghei) has shown strong recruitment to the cortical meninges of immune cells, including neutrophils, T cells, and putative dendritic cells and macrophages. Without special labeling, the boundaries between the dura mater, the leptomeninx, and the parenchyma are not directly visualized in intravital two-photon microscopy, but other landmarks and characteristics, which we illustrate, allow the researcher to identify the compartment being imaged. While most infectious meningitides are localized mainly in the dura mater, others involve recruitment of immune cells to the leptomeninx.


1-Year Results in Patients Undergoing Transcatheter Aortic Valve Replacement With Failed Surgical Bioprostheses.
This study evaluated the safety and effectiveness of self-expanding transcatheter aortic valve replacement (TAVR) in patients with surgical valve failure (SVF). Self-expanding TAVR is superior to medical therapy for patients with severe native aortic valve stenosis at increased surgical risk. The CoreValve U.S. Expanded Use Study was a prospective, nonrandomized study that enrolled 233 patients with symptomatic SVF who were deemed unsuitable for reoperation. Patients were treated with self-expanding TAVR and evaluated for 30-day and 1-year outcomes after the procedure. An independent core laboratory was used to evaluate serial echocardiograms for valve hemodynamics and aortic regurgitation. SVF occurred through stenosis (56.4%), regurgitation (22.0%), or a combination (21.6%). A total of 227 patients underwent attempted TAVR and successful TAVR was achieved in 225 (99.1%) patients. Patients were elderly (76.7 ± 10.8 years), had a Society of Thoracic Surgeons Predicted Risk of Mortality score of 9.0 ± 6.7%, and were severely symptomatic (86.8% New York Heart Association functional class III or IV). The all-cause mortality rate was 2.2% at 30 days and 14.6% at 1 year; major stroke rate was 0.4% at 30 days and 1.8% at 1 year. Moderate aortic regurgitation occurred in 3.5% of patients at 30 days and 7.4% of patients at 1 year, with no severe aortic regurgitation. The rate of new permanent pacemaker implantation was 8.1% at 30 days and 11.0% at 1 year. The mean valve gradient was 17.0 ± 8.8 mm Hg at 30 days and 16.6 ± 8.9 mm Hg at 1 year. Factors significantly associated with higher discharge mean aortic gradients were surgical valve size, stenosis as modality of SVF, and presence of surgical valve prosthesis patient mismatch (all p < 0.001). Self-expanding TAVR in patients with SVF at increased risk for surgery was associated with a low 1-year mortality and major stroke rate, significantly improved aortic valve hemodynamics, and low rates of moderate and no severe residual aortic regurgitation, with improved quality of life.


Chylothorax and pseudochylothorax.
Chylothorax is the occurrence of chylus (lymph) in the pleura due to damage to the thoracic duct. There is a high content of triglycerides, and chylomicrons can be seen. It is usually right-sided, since most of the duct is within the right hemithorax. With damage at the level of the aorta, the chyle will appear on the left. Traumatic rupture occurs after accidents or surgery. Of nontraumatic causes, the most common is a malignant lymphoma. Computed tomography (CT) scan of the thorax and upper abdomen should be performed. Lymphography can show where the leakage or blockage is situated. With repeated drains, large amounts of fat, proteins, and lymphocytes are lost. Treatment is with low-fat diet or parenteral nutrition to decrease the amount of chyle, but chemical pleurodesis or ligation of the thoracic duct, usually via thoracoscopy, is often necessary. Pseudochylothorax (cholesterol pleurisy) occurs with long-standing fluid in a fibrotic pleura. The fluid has a high content of cholesterol but no triglycerides or chylomicrons. In both conditions, the pleural fluid is thick, opalescent, whitish or the colour of cafe-au-lait.


Surfactin from Bacillus subtilis attenuates ambient air particulate matter-promoted human oral cancer cells metastatic potential.
Recently, many studies have indicated that ambient air particulate matter (PM) can increase the risk of oral cancer. The most common malignant tumor in the oral cavity is oral squamous cell carcinoma (OSCC). Usually, cancer cell migration/invasion is the most important cause of cancer mortality. Matrix metalloproteinase-2 (MMP-2) and MMP-9 have been shown to play important roles in regulating metastasis and the tumor microenvironment. Here, we studied the anti-cancer effects of surfactin, a cyclic lipopeptide generated by Bacillus subtilis, on cancer cell migration and invasion. Surfactin suppressed PM-promoted cell migration and invasion and colony formation of SCC4 and SCC25 human oral squamous cell carcinoma cell lines. We observed that PM induced MMP-2 and MMP-9 expression, which was inhibited by surfactin. Transfection with p65, p50, c-Jun, c-Fos, p85, p110, Akt, mammalian target of rapamycin (mTOR), or interleukin-6 (IL-6) siRNA markedly inhibited PM-induced MMP-2 and MMP-9 expression. Moreover, surfactin could reduce Akt, mTOR, p65, and c-Jun activation and IL-6 secretion induced by PM. Finally, we proved that transfection with Akt, p65, or c-Jun siRNA significantly inhibited PM-induced IL-6 release. Taken together, these results suggest that surfactin functions as a suppressor of PM-induced MMP2/9-dependent oral cancer cell migration and invasion by inhibiting the activation of phosphoinositide 3-kinase (PI3K)/Akt/mTOR and PI3K/Akt/nuclear factor-κB (NF-κB) and activator protein-1 (AP-1)/IL-6 signaling pathways.


Beverage Intake During Pregnancy and Childhood Adiposity.
To examine associations of sugar sweetened beverages (SSBs) and other beverage intake during pregnancy with adiposity in midchildhood (median age of 7.7 years). We studied 1078 mother-child pairs in Project Viva, a prospective prebirth cohort study. Exposures were sugary and nonsugary beverage intake assessed in the first and second trimesters of pregnancy by using a food frequency questionnaire. Main outcome measures were offspring overall adiposity (BMI z score, fat mass index [FMI, kg/m2] from dual-energy radiograph absorptiometry, and sum of subscapular [SS] and triceps [TR] skinfold thicknesses) and central adiposity (SS:TR ratio and waist circumference). In the second trimester, mean (SD) SSB intake was 0.6 (0.9) servings per day. Thirty-two percent of mothers were multiracial or people of color, 68% were college graduates, and 10% smoked during pregnancy. In midchildhood, mean (SD) BMI z score was 0.38 (1.00), and the FMI was 4.4 (1.9) kg/m2. In multivariable models adjusted for multiple maternal and child covariates, each additional serving per day of SSB was associated with higher BMI z scores (0.07 U; 95% confidence interval [CI]: -0.01 to 0.15), FMI (0.15 kg/m2; 95% CI: -0.01 to 0.30), SS + TR (0.85 mm; 95% CI: 0.06 to 1.64), and waist circumference (0.65 cm; 95% CI: 0.01 to 1.28). Stratified models suggested that the associations were due primarily to maternal, not child, SSB intake and to sugary soda rather than fruit drinks or juice. We did not find differences between boys and girls, nor did we observe the effects of sugar-free soda or of first-trimester beverage intake. Higher SSB intake during the second trimester of pregnancy was associated with greater adiposity in midchildhood.


Competitive and cooperative functioning of the anterior and posterior promoter elements of an Alu family repeat.
Similar to tRNA genes and the VAI gene, the Alu family repeats are transcribed by RNA polymerase III and contain a split intragenic promoter. Results of our previous studies have shown that when the anterior, box A-containing promoter element (5'-Pu-Pu-Py-N-N-Pu-Pu-Py-G-G-3' in which Pu is any purine, Py is any pyrimidine, and N is any nucleotide) of a human Alu family repeat is deleted, the remaining box B-containing promoter element (5'-G-A/T-T-C-Pu-A-N-N-C-3') is still capable of directing weak transcriptional initiation at approximately 70 base pairs (bp) upstream from the box B sequence. This is different from the tRNA genes in which the box A-containing promoter element plays the major role in the positioning of the transcriptional initiation site(s). To account for this difference, we first carried out competition experiments in which we show that the posterior element of the Alu repeat competes with the VAI gene effectively for the transcription factor C in HeLa cell extracts. We then constructed a series of contraction and expansion mutants of the Alu repeat promoter in which the spacing between boxes A and B was systematically varied by molecular cloning. In vitro transcription of these clones in HeLa cell extracts was analyzed by RNA gel electrophoresis and primer extension mapping. We show that when the box A and box B promoter sequences are separated by 47 to 298 bp, the transcriptional initiation sites remain 4 to 5 bp upstream from box A. However, this positioning function by the box A-containing promoter element was lost when the spacing was shortened to only 26 bp or increased to longer than 600 bp. Instead, transcriptional initiation occurred approximately 70 bp upstream from box B, similar to that in the clones containing only the box B promoter element. All the mutant clones were transcribed less efficiently than was the wild type. An increase in the distance between boxes A and B also activated a second box A-like element within the Alu family repeat. We compare these results with the results of tRNA gene studies. We also discuss this comparison in terms of the positioning function of the split class III promoter elements and the evolutionary conservation of the spacing between the two promoter elements for optimum transcriptional efficiency.


Analysis and improvement of fitness exploitation in XCS: bounding models, tournament selection, and bilateral accuracy.
The evolutionary learning mechanism in XCS strongly depends on its accuracy-based fitness approach. The approach is meant to result in an evolutionary drive from classifiers of low accuracy to those of high accuracy. Since, given inaccuracy, lower specificity often corresponds to lower accuracy, fitness pressure most often also results in a pressure towards higher specificity. Moreover, fitness pressure should cause the evolutionary process to be innovative in that it combines low-order building blocks of lower accurate classifiers, to higher-order building blocks with higher accuracy. This paper investigates how, when, and where accuracy-based fitness results in successful rule evolution in XCS. Along the way, a weakness in the current proportionate selection method in XCS is identified. Several problem bounds are derived that need to be obeyed to enable proper evolutionary pressure. Moreover, a fitness dilemma is identified that causes accuracy-based fitness to be misleading. Improvements are introduced to XCS to make fitness pressure more robust and overcome the fitness dilemma. Specifically, (1) tournament selection results in a much better fitness-bias exploitation, and (2) bilateral accuracy prevents the fitness dilemma. While the improvements stand for themselves, we believe they also contribute to the ultimate goal of an evolutionary learning system that is able to solve decomposable machine-learning problems quickly, accurately,and reliably. The paper also contributes to the further understanding of XCS in general and the fitness approach in XCS in particular.


Quality and safety in the transitional care of the elderly (phase 2): the study protocol of a quasi-experimental intervention study for a cross-level educational programme.
Transitional care and patient handover are important areas to ensure quality and safety in elderly healthcare services. Previous studies showed that healthcare professionals have little knowledge of the setting they are transferring patients to and a limited understanding of roles and functions; these constitute barriers to effective communication and shared care responsibilities across levels of care. The main objective is to implement a cross-level education-based intervention programme with healthcare professionals aimed at (1) increasing professionals' awareness and competencies about quality and safety in the transitional care of the elderly; (2) creating a discussion platform for knowledge exchange and learning across levels and units of care and (3) improving patient safety culture, in particular, in transitional care. A quasi-experimental control group study design with an intervention group and a control group; this includes a pretest, post-test and 1-year follow-up test assessment of patient safety culture. Qualitative data will be collected during the intervention programme and between the measurements. The study design will be beneficial for addressing the effects of the cross-level educational intervention programme on reports of patient safety culture and for addressing the feasibility of the intervention measures. The study has been approved by the Regional Committees for Medical and Health Research Ethics in Norway, Ref. No. 2011/1978. The study is based on informed written consent; informants can withdraw from the study at any point in time. The results will be disseminated at research conferences, in peer review journals and through public presentations outside the scientific community.


Observation of Correlated Azimuthal Anisotropy Fourier Harmonics in pp and p+Pb Collisions at the LHC.
The azimuthal anisotropy Fourier coefficients (v_{n}) in 8.16 TeV p+Pb data are extracted via long-range two-particle correlations as a function of the event multiplicity and compared to corresponding results in pp and PbPb collisions. Using a four-particle cumulant technique, v_{n} correlations are measured for the first time in pp and p+Pb collisions. The v_{2} and v_{4} coefficients are found to be positively correlated in all collision systems. For high-multiplicity p+Pb collisions, an anticorrelation of v_{2} and v_{3} is observed, with a similar correlation strength as in PbPb data at the same multiplicity. The new correlation results strengthen the case for a common origin of the collectivity seen in p+Pb and PbPb collisions in the measured multiplicity range.


Marked cytoreduction of a lymphocyte-rich mediastinal thymoma with neoadjuvant chemotherapy in a cat.
A 15-year-old neutered female domestic shorthair cat presented with lethargy and acute-onset dyspnoea. Thoracic computed tomography (CT) revealed a large, cranial mediastinal mass with an estimated volume of 180.7 cm3. Chemotherapy consisting of dexamethasone followed by L-asparaginase, prednisolone, vincristine and doxorubicin was commenced owing to the severity of disease and initial possibility of lymphoma. A diagnosis of lymphocyte-rich thymoma was made based upon histological examination, positive pancytokeratin staining, variable lymphocyte CD3 expression and T cell receptor gamma polyclonality. Thoracic CT performed 35 days after the commencement of chemotherapy showed a marked reduction in the size of the mass, with an estimated volume of 9.4 cm3. A median sternotomy and thymectomy were performed. No clinical signs have recurred 34 months after surgery. The response to chemotherapy in this case was unusual, and is likely associated with the high non-neoplastic lymphoid component of the mass. The case demonstrates that preoperative chemotherapy can be used to reduce thymoma volume prior to surgery, potentially decreasing anaesthetic risk.


First Report of Aspergillus niger Causing Postharvest Fruit Rot of Cherry in the Prefectures of Imathia and Pella, Northern Greece.
In June 2011, symptoms of postharvest rot were observed on approximately 3% of all cherries collected from commercial orchards of cultivars Lapen and Ferrovia in the prefectures of Imathia and Pella (northern Greece). Fruit were harvested in a timely manner to avoid overripeness. No wounds or other predisposing injuries were observed on the infected fruits. Lesions enlarged rapidly and separated easily from healthy tissue when pressure was applied. Infected tissues were pale and water soaked and the associated fungal spores were dark and powdery and easily liberated when mature. The fungus grew rapidly and produced black colonies on acidified potato dextrose agar (2.5 ml of 85% lactic acid per liter of nutrient medium) after 5 days at 24°C. Identification of the pathogen was based on morphological characteristics (1). The conidial head was radiate, vesicles were nearly spherical and covered with metulae and phialides (biseriate). Conidia were globose (3 to 5 μm in diameter) and usually very rough with irregular ridges, bars, and verrucae. Koch's postulates were completed in the laboratory by inoculating mature cherry fruits (cv. Lapen). The fruits were surface sterilized by dipping in 10% chloride bleach solution, allowed to dry in a laminar flow hood, and wounded with a sharp glass rod that was 2 mm in diameter. A 40-μl drop of a suspension containing 20,000 conidia per ml of water was placed on each wound. There were 20 inoculated and 20 control fruits (similarly wounded and inoculated with a 40-μl drop of sterile distilled water) in a randomized design and incubated at 24 to 26°C for 6 days. Koch's postulates were satisfied after reisolating the fungus from inoculated fruit that developed symptoms similar to those observed on fruit collected from orchards. Control fruits did not show any symptom of the disease. To our knowledge, this is the first report of the occurrence of Aspergillus niger as the causal agent of postharvest rots of cherries in Greece. Postharvest fruit rots caused by A. niger have been reported in cherry orchards of other countries around the world (2). Because this disease causes postharvest rots of cherry fruits, measures may need to be implemented to manage the pathogen. References: (1) M. A. Klich. Page 12 in: Identification of Common Aspergillus Species. Centraalbureau Voor Schimmelcultures, Utrecht, the Netherlands, 2002. (2) A. Valiuskaite et al. Phytopathol. Pol. 35:197, 2005.


Direct-Contact Microelectrical Measurement of the Electrical Resistivity of a Solid Electrolyte Interface.
Because of its effectiveness in blocking electrons, the solid electrolyte interface (SEI) suppresses decomposition reactions of the electrolyte and contributes to the stability and reversibility of batteries. Despite the critical role of SEI in determining the properties of batteries, the electrical properties of SEI layers have never been measured directly. In this paper, we present the first experimental results of the electrical resistivity of a LiF-rich SEI layer measured using a direct-contact microelectrical device mounted in an electron microscope. Measurements show that the SEI layer exhibits high electrical resistivity (2.3 × 105 Ω·m), which is comparable with those of typical insulating materials. Furthermore, a combined technique of advanced analyses and first-principles calculations show that the SEI layer is mainly composed of amorphous LiF and a minute nanocrystalline Li2CO3 compound. The electronic origin responsible for the high resistivity of the SEI layer is elucidated by calculating the band structures of various Li xF compounds and interpreting their effects on the resistivity. This study explains why SEI can prevent the degradation of electrode materials and consumption of Li ions in the electrolyte and thus can be viewed as a stepping stone for developing highly stable and reversible batteries.


Second consensus statement on the diagnosis of multiple system atrophy.
A consensus conference on multiple system atrophy (MSA) in 1998 established criteria for diagnosis that have been accepted widely. Since then, clinical, laboratory, neuropathologic, and imaging studies have advanced the field, requiring a fresh evaluation of diagnostic criteria. We held a second consensus conference in 2007 and present the results here. Experts in the clinical, neuropathologic, and imaging aspects of MSA were invited to participate in a 2-day consensus conference. Participants were divided into five groups, consisting of specialists in the parkinsonian, cerebellar, autonomic, neuropathologic, and imaging aspects of the disorder. Each group independently wrote diagnostic criteria for its area of expertise in advance of the meeting. These criteria were discussed and reconciled during the meeting using consensus methodology. The new criteria retain the diagnostic categories of MSA with predominant parkinsonism and MSA with predominant cerebellar ataxia to designate the predominant motor features and also retain the designations of definite, probable, and possible MSA. Definite MSA requires neuropathologic demonstration of CNS alpha-synuclein-positive glial cytoplasmic inclusions with neurodegenerative changes in striatonigral or olivopontocerebellar structures. Probable MSA requires a sporadic, progressive adult-onset disorder including rigorously defined autonomic failure and poorly levodopa-responsive parkinsonism or cerebellar ataxia. Possible MSA requires a sporadic, progressive adult-onset disease including parkinsonism or cerebellar ataxia and at least one feature suggesting autonomic dysfunction plus one other feature that may be a clinical or a neuroimaging abnormality. These new criteria have simplified the previous criteria, have incorporated current knowledge, and are expected to enhance future assessments of the disease.


Comprehensive discharge planning with postdischarge support for older patients with congestive heart failure: a meta-analysis.
Comprehensive discharge planning plus postdischarge support may reduce readmission rates for older patients with congestive heart failure (CHF). To evaluate the effect of comprehensive discharge planning plus postdischarge support on the rate of readmission in patients with CHF, all-cause mortality, length of stay (LOS), quality of life (QOL), and medical costs. We searched MEDLINE (1966 to October 2003), the Cochrane Clinical Trials Register (all years), Social Science Citation Index (1992 to October 2003), and other databases for studies that described such an intervention and evaluated its effect in patients with CHF. Where possible we also contacted lead investigators and experts in the field. We selected English-language publications of randomized clinical trials that described interventions to modify hospital discharge for older patients with CHF (mean age > or =55 years), delineated clearly defined inpatient and outpatient components, compared efficacy with usual care, and reported readmission as the primary outcome. Two authors independently reviewed each report, assigned quality scores, and extracted data for primary and secondary outcomes in an unblinded standardized manner. Eighteen studies representing data from 8 countries randomized 3304 older inpatients with CHF to comprehensive discharge planning plus postdischarge support or usual care. During a pooled mean observation period of 8 months (range, 3-12 months), fewer intervention patients were readmitted compared with controls (555/1590 vs 741/1714, number needed to treat = 12; relative risk [RR], 0.75; 95% confidence interval [CI], 0.64-0.88). Analysis of studies reporting secondary outcomes found a trend toward lower all-cause mortality for patients assigned to an intervention compared with usual care (RR, 0.87; 95% CI, 0.73-1.03; n = 14 studies), similar initial LOS (mean [SE]: 8.4 [2.5] vs 8.5 [2.2] days, P =.60; n = 10), greater percentage improvement in QOL scores compared with baseline scores (25.7% [95% CI, 11.0%-40.4%] vs 13.5% [95% CI, 5.1%-22.0%]; n = 6, P =.01), and similar or lower charges for medical care per patient per month for the initial hospital stay, administering the intervention, outpatient care, and readmission (-359 dollars [95% CI, -763 dollars to 45 dollars]; n = 4, P =.10 for non-US trials and -536 dollars [95% CI, -956 dollars to -115 dollars]; n = 4, P =.03, for US trials). Comprehensive discharge planning plus postdischarge support for older patients with CHF significantly reduced readmission rates and may improve health outcomes such as survival and QOL without increasing costs.


Maternal stress in nonverbal learning disorder: a comparison with reading disorder.
Maternal stress was assessed in mothers of children ages 8 to 11 years with learning disorders (LD). Age-, gender-, and IQ-matched children with reading disorders (RD; n = 31), children with nonverbal learning disorders (NVLD; n = 21), and typically developing control participants (n = 23) participated. Mothers of children with LD reported higher levels of stress, although the specific nature of the stress appeared to vary by disorder subtype: Mothers of children with RD reported higher levels of general distress, whereas mothers of children with NVLD reported higher levels of dysfunctional interactions with their child. The severity of the LD was strongly associated with maternal stress in the sample of children with NVLD but not in the sample of children with RD. In the sample of children with NVLD, the best predictors of maternal stress were 2 variables associated with the child: The lower the child's Performance IQ, the higher the level of maternal stress. Furthermore, the level of the child's internalizing symptoms was also a strong predictor of maternal stress in the sample of children with NVLD. In our sample of children with RD, the 3 best predictors were all variables associated with the mother, including her age, her level of reported psychiatric symptoms, and her overall level of social support.


Neurosurgical treatment of progressive posthemorrhagic ventricular dilation in preterm infants: a 10-year single-institution study.
Intraventricular hemorrhage (IVH) and progressive posthemorrhagic ventricular dilation (PPHVD) may result in significant neurological morbidity in preterm infants. At present, there is no consensus regarding the optimal timing or type of neurosurgical procedure to best treat PPHVD. Conflicting data exist regarding the relative risks and benefits of two commonly used temporizing neurosurgical procedures (TNPs), ventricular access devices ([VADs] or ventricular reservoirs) versus ventriculosubgaleal (VSG) shunts. This study was designed to address this issue. This is a single-center, 10-year retrospective review of all preterm infants admitted to the St. Louis Children's Hospital neonatal intensive care unit (NICU) with Papile Grade III-IV IVH. The development of PPHVD and the requirement for and type of TNP were recorded. Rates of TNP complication, ventriculoperitoneal (VP) shunt implantation, shunt infection, and mortality rates were used to compare the efficacy and limitations of each TNP type. Over this 10-year interval, 325 preterm infants with Grade III-IV IVH were identified, with trends showing an increasing number of affected infants annually, and an increasing number of TNPs were required annually. Ninety-five (29.2%) of the 325 infants underwent a TNP for PPHVD (65 VADs, 30 VSG shunts). The rate of permanent VP shunt implantation for all TNPs was 72.6% (69 of 95 infants). Forty-nine (75.4%) of the 65 infants treated with VADs and 20 (66.7%) of the 30 treated with VSG shunts required VP shunts (p = 0.38). There was no statistical difference between VAD or VSG shunt with regard to TNP-related infection (p = 0.57), need for TNP revision (p = 0.16), subsequent shunt infection (p = 0.77), shunt revision rate (p = 0.58), or mortality rate (p = 0.24). Rates of IVH and PPHVD observed at the authors' center have increased over time. In contrast to recent literature, the results from the current study did not demonstrate a difference in complication rate or requirement for permanent VP shunt placement between VADs and VSG shunts. Definitive conclusions will require a larger, prospective trial.


Antibiotic Practice Patterns for Extremity Wound Infections among Blast-Injured Subjects.
We examined antibiotic management of combat-related extremity wound infections (CEWI) among wounded U.S. military personnel (2009-2012). Patients were included if they sustained blast injuries, resulting in ≥1 open extremity wound, were admitted to participating U.S. hospitals, developed a CEWI (osteomyelitis or deep soft-tissue infections) within 30 days post-injury, and received ≥3 days of relevant antibiotic (s) for treatment. Among 267 patients, 133 (50%) had only a CEWI, while 134 (50%) had a CEWI plus concomitant non-extremity infection. In the pre-diagnosis period (4-10 days prior to CEWI diagnosis), 95 (36%) patients started a new antibiotic with 28% of patients receiving ≥2 antibiotics. During CEWI diagnosis week (±3 days of diagnosis), 209 (78%) patients started a new antibiotic (71% with ≥2 antibiotics). In the week following diagnosis (4-10 days after CEWI diagnosis), 121 (45%) patients started a new antibiotic with 39% receiving ≥2 antibiotics. Restricting to ±7 days of CEWI diagnosis, patients commonly received two (35%) or three (27%) antibiotics with frequent combinations involving carbapenem, vancomycin, and fluoroquinolones. Substantial variation in antibiotic prescribing patterns related to CEWIs warrants development of combat-related clinical practice guidelines beyond infection prevention, to include strategies to reduce the use of unnecessary antibiotics and improve stewardship.


Does nutrition play a role in the prevention and management of sarcopenia?
There is a growing body of evidence that links nutrition to muscle mass, strength and function in older adults, suggesting that it has an important role to play both in the prevention and management of sarcopenia. This review summarises the discussions of a working group [ESCEO working group meeting 8th September 2016] that met to review current evidence and to consider its implications for preventive and treatment strategies. The review points to the importance of 'healthier' dietary patterns that are adequate in quality in older age, to ensure sufficient intakes of protein, vitamin D, antioxidant nutrients and long-chain polyunsaturated fatty acids. In particular, there is substantial evidence to support the roles of dietary protein and physical activity as key anabolic stimuli for muscle protein synthesis. However, much of the evidence is observational and from high-income countries. Further high-quality trials, particularly from more diverse populations, are needed to enable an understanding of dose and duration effects of individual nutrients on function, to elucidate mechanistic links, and to define optimal profiles and patterns of nutrient intake for older adults.


Chemical and carbon-13 cross-polarization magic-angle spinning nuclear magnetic resonance characterization of logyard fines from British Columbia.
Phasing out beehive burners and rising costs for landfilling have increased the need to widen options for utilization of the smaller size fractions of woody wastes generated during log handling and sawmilling in British Columbia. We characterized several size classes of logyard fines up to 16 mm sampled from coastal and interior operations. Total C, total N, ash, and condensed tannin concentrations were consistent with properties derived largely from wood, with varying proportions of bark and mixing with mineral soil. Especially for < 3-mm fractions, the latter resulted in high ash contents that would make them unsuitable for fuel. Solid-state 13C cross-polarization magic-angle spinning (CPMAS) nuclear magnetic resonance (NMR) spectra were consistent with the chemical data, with high O-alkyl intensity and similarity to naturally occurring woody forest floor; no samples were high in aromatic or phenolic C. Aqueous extracts of two < 16-mm fines, which accounted for only a small proportion of the total C, were enriched in alkyl C and had low or undetectable tannins. Application to forest sites might cause short-term immobilization of N, but also might include possible longer-term benefits from reduction of N loss after harvesting and restoration of soil organic matter in degraded sites.


The interaction of lipodepsipeptide toxins from Pseudomonas syringae pv. syringae with biological and model membranes: a comparison of syringotoxin, syringomycin, and two syringopeptins.
Pseudomonas syringae pv. syringae produces two groups of cyclic lipodepsipeptides (LDPs): the nona-peptides syringomycins, syringostatins, and syringotoxin (ST), and the more complex syringopeptins composed of either 22 or 25 amino acid residues (SP22 and SP25). Both classes of peptides significantly contribute to bacterial pathogenesis and their primary target of action seems to be the plasma membrane. We studied and compared the activity of some members of these two classes of LDPs on red blood cells and on model membranes (monolayers and unilamellar vesicles). All peptides induced red blood cell hemolysis. The mechanism was apparently that of a colloid-osmotic shock caused by the formation of pores, as it could be prevented by osmoticants of adequate size. Application of the Renkin equation indicated a radius of approximately 1 nm for the lesions formed by syringopeptins SP22A and SP25A, whereas those formed by syringomycin E (SRE) had a variable, dose-dependent size ranging from 0.7 up to 1.7 nm. All tested LDPs displayed surface activity, forming peptide monolayers with average molecular areas of 1.2 nm2 (SRE), 1.5 nm2 (SP22A), and 1.3 nm2 (SP25A). They also partitioned into preformed lipid monolayers occupying molecular areas that ranged from 0.6 to 1.7 nm2 depending on the peptide and the lipid composition of the film. These LDPs formed channels in lipid vesicles as indicated by the release of an entrapped fluorescent dye (calcein). The extent of permeabilization was dependent on the concentration of the peptide and the composition of the lipid vesicles, with a preference for those containing a sterol. From the dose dependence of the permeabilization it was inferred that LDPs increased membrane permeability by forming oligomeric channels containing from four to seven monomers. On average, syringopeptin oligomers were smaller than SRE and ST oligomers.


Characterization of the first beta-class carbonic anhydrase from an arthropod (Drosophila melanogaster) and phylogenetic analysis of beta-class carbonic anhydrases in invertebrates.
The beta-carbonic anhydrase (CA, EC 22.214.171.124) enzymes have been reported in a variety of organisms, but their existence in animals has been unclear. The purpose of the present study was to perform extensive sequence analysis to show that the beta-CAs are present in invertebrates and to clone and characterize a member of this enzyme family from a representative model organism of the animal kingdom, e.g., Drosophila melanogaster. The novel beta-CA gene, here named DmBCA, was identified from FlyBase, and its orthologs were searched and reconstructed from sequence databases, confirming the presence of beta-CA sequences in 55 metazoan species. The corresponding recombinant enzyme was produced in Sf9 insect cells, purified, kinetically characterized, and its inhibition was investigated with a series of simple, inorganic anions. Holoenzyme molecular mass was defined by dynamic light scattering analysis and gel filtration, and the results suggested that the holoenzyme is a dimer. Double immunostaining confirmed predictions based on sequence analysis and localized DmBCA protein to mitochondria. The enzyme showed high CO2 hydratase activity, with a kcat of 9.5 x 105 s-1 and a kcat/KM of 1.1 x 108 M-1s-1. DmBCA was appreciably inhibited by the clinically-used sulfonamide acetazolamide, with an inhibition constant of 49 nM. It was moderately inhibited by halides, pseudohalides, hydrogen sulfide, bisulfite and sulfate (KI values of 0.67 - 1.36 mM) and more potently by sulfamide (KI of 0.15 mM). Bicarbonate, nitrate, nitrite and phenylarsonic/boronic acids were much weaker inhibitors (KIs of 26.9 - 43.7 mM). The Drosophila beta-CA represents a highly active mitochondrial enzyme that is a potential model enzyme for anti-parasitic drug development.


Correlation of hepatitis C RNA and serum alanine aminotransferase in hepatitis B and C seronegative healthy blood donors.
Historically, serum alanine transaminase (ALT) has been used as a surrogate marker in the detection of hepatitis viruses in blood donors. With the availability of newer sensitive technologies for the detection of seroconversion, the value of ALT becomes questionable but continues to be used for this purpose with subsequent discarding of ALT elevated blood units. The present study aims to evaluate the significance and cost effectiveness of ALT as a surrogate marker for hepatitis C virus infection in healthy asymptomatic blood donors who were serologically negative. The study was conducted at clinical laboratory of a tertiary care hospital for a period of one year from November 2006 to October 2007. All donors were screened serologically for hepatitis B, C and HIV I and II, syphilis and malaria and those tested positive were excluded from further evaluation. Gender-wise reference ranges and minimal and markedly raised results for ALT (described respectively as one and two folds increase above reference range) were defined and, accordingly, donors were grouped into three. Two hundred seronegative blood donors were randomly selected from all three groups of ALT results and tested for hepatitis C nucleic acid through Amplicor HCV RNA test. The cost of discarding an ALT -only elevated blood unit was also assessed. During the study period, 25117 subjects donated blood. EIGHT HUNDRED AND RESULTS: Seventy two donors (3.4%) were positive for one or more serological tests. ALT of all donors ranged from 0-1501 U/L (Mean +/- SD; 33.4 +/- 25.45 U/L). The donors seronegative for all disease markers were 24245 (96.6%). Of these, 21164 (87.2%) donors had their ALT within reference range while 2874 (11.8%) and 207 (0.8%) of donors had minimal and markedly elevated results. Thus, 621 blood bags (red cells, platelets and plasma) costing $ 39200.0 were discarded based on ALT results alone. Of 200 seronegative donors evaluated for hepatitis C nucleic acid, only one within markedly elevated ALT levels was found to be positive. The present work did not support a positive association between hepatitis C virus nucleic acid and elevated ALT in healthy serologically negative blood donors. We did not find serum ALT testing in donors as cost effective strategy for detection of hepatitis C virus ribonucleic acid. As the number of samples tested by us was small we suggest further work to evaluate the value of ALT levels in serologically negative donors in association with hepatitis C antigen and NAT testing to elucidate the true burden of disease in geographical regions where hepatitis C is endemic and voluntary blood donation is sparse.


Morphologic evaluation of the thoracic vertebrae for safe free-hand pedicle screw placement in adolescent idiopathic scoliosis: a CT-based anatomical study.
To evaluate the morphologic characteristics of the thoracic pedicle with regard to safe free-hand thoracic pedicle screw placement, based on multi-planar reconstruction CT images. Thirty adolescent idiopathic scoliosis (AIS) patients who had undergone posterior corrective surgery for major thoracic curve were included in this study. Reconstructed CT axial images at each thoracic vertebra were used to measure: (1) the shortest distance from an entry point to the ventral cortex of the lamina (critical distance), and (2) the distance from the entry point to the tangent of the spinal canal at the medial wall of the pedicle (safe distance). The critical length was defined as the distance between the critical distance and the safe distance. The distance from the entry point to the medial wall breach site (breach distance) was measured on post-operative CT images. The mean critical distance was 9.3 ± 1.1 mm. The critical distance of vertebrae from different levels was relatively constant, between 8.1 and 10.1 mm. The mean safe distance was 15.2 ± 1.3 mm. The safe distance of vertebrae from different levels was also relatively constant, between 14.5 and 16 mm. The mean critical length was 5.9 ± 1.0 mm. The critical length of vertebrae between T3 and T12 was relatively constant, ranging from 5 to 6.5 mm. The mean breach distance was 12.3 ± 1.3 mm and the each breach always recognized between the critical distance and the safe distance. The risk of pedicle medial wall perforation increases as the pedicle probe advances beyond the critical distance of 8-10 mm from the entry point, while it decreases entering into the safe distance at 14.5-16 mm. These parameters were relatively constant even in the most rotated vertebrae at T9 or those with the narrowest pedicle at T7 or T4.


Overview of disc arthroplasty-past, present and future.
Degenerative disc disease is one of the most frequent spinal disorders. The anatomy and the biomechanics of the intervertebral disc are very complex, and the pathomechanics of its degeneration are poorly understood. Despite this complexity and uncertainty, great advances have been made in the field of disc replacement technology, with promising results. Difficulties are continuously being encountered, but careful analysis of the results and intensive research and development will assist in countering these problems. There are approximately 40 clinical reports in the literature describing various aspects of randomised controlled trials involving intervertebral disc arthroplasty. However, the majority of these publications do not provide reliable information, in that they give only interim results and/or the results from just one of the many centres in multicentre studies. Such publications must be interpreted with caution, since they do not always represent the results of the whole study population and may hence be underpowered. We identified six randomised controlled trials that compared the final clinical outcomes of disc arthroplasty and spinal fusion. The present systematic review attempts to give an overview of the current status of disc arthroplasty.


Mortality salience modulates cortical responses to painful somatosensory stimulation: Evidence from slow wave and delta band activity.
Social psychology studies show that awareness of one's eventual death profoundly influences human cognition and behaviour by inducing defensive reactions against end-of-life-related anxiety. Much less is known about the impact of reminders of mortality on brain activity. Here we tested whether reminders of mortality can induce a modulation of the slow electroencephalographic activity triggered by somatosensory nociceptive or auditory threatening stimulation and if this modulation is related to mood and anxiety as well as personality traits. We found a specific slow wave (SW) modulation only for nociceptive stimulation and only following mortality salience induction (compared to reminders of an important failed exam). The enhancement of SW negativity at the scalp vertex was associated with increased state anxiety and negative mood, whereas higher self-esteem was associated with reduced SW amplitude. In addition, mortality salience was linked to an increased amplitude of frontal delta band, which was correlated also with increased positive mood and higher self-esteem. The results indicate that SW and delta spectral activity may represent both proximal and distal defences associated with reminders of death and that neurophysiological correlates of somatosensory representation of painful and threatening stimuli may be useful for existential neuroscience studies.


The electrochemical proton gradient and its influence on citrate uptake in tonoplast vesicles of Hevea brasiliensis.
The relationship between the electrochemical proton gradient, Δ μH+ (-) , and citrate transport has been studied in tonoplast vesicles from Hevea brasiliensis (the rubber tree). Vesicles were generated from lyophilized samples of fresh vacuoles obtained from the latex sap. Methylamine was used to measure intravesicular pH and lipophilic ions to determine the electrical potential difference (ΔΨ) across the tonoplast. When incubated at pH 7.5 in the absence of ATP, the tonoplast vesicles showed a ΔpH of 0.6 units (interior acid) and a ΔΨ of about-100 mV (interior negative). This potential is thought to be made up of contributions from an H(+) diffusion potential, diffusion potentials from other cations and a Donnan potential arising from the presence of internal citrate. In the presence of 5 mol m(-3) MgATP the ΔpH was increased to about 1.0 unit and the ΔΨ to about-10 mV. Under these conditions the proton-motive force (Δ p Δ μH+ (-) /F) became positive and reached +50 mV. These effects were specific to MgATP (ADP and Mg(2+) having no significant effect) and were prevented by the protonophore p-trifluoromethoxycarbonylcyanidephenylhydrazone (FCCP). Citrate uptake by the vesicles was markedly stimulated by MgATP; ADP and Mg(2+) again had no effect. Nigericin greatly increased ΔpH and this was associated with a large increase in citrate accumulation. The results indicate that the vesicle membrane possesses a functional H(+)-translocating ATPase. The Δ μH+ (-) generated by this ATPase can be used to drive citrate uptake into the vesicles. The properties of the tonoplast vesicles are compared with those of the fresh latex vacuoles.


OCT imaging of aorto-coronary vein graft pathology modified by external stenting: 1-year post-surgery.
The Venous External Support Trial (VEST) evaluated whether a novel external stent attenuated saphenous vein graft (SVG) disease assessed with intravascular ultrasound 1 year following coronary artery bypass graft (CABG) surgery. This sub-study assessed SVGs with and without external stenting using optical coherence tomography (OCT). The aim of this study was to accurately compare quantitative and qualitative features of SVGs with and without a novel external stent using OCT. Twenty-four of 30 patients (65 ± 8 years) enrolled in VEST underwent coronary angiography with OCT imaging using a non-occlusive technique. Quantitative analysis of lumen area was performed in one frame every 10 mm along the length of the graft, from distal to proximal anastomosis, and pathological features within the lumen were noted. Mean cross-sectional area was greater in unstented vs. stented grafts (8.4 ± 3 vs. 7.6 ± 2.7 mm; P = 0.005). The lumen of the stented grafts was more homogeneous (difference between maximum and minimum lumen diameter was significantly smaller in stented compared with unstented grafts, 0.28 ± 0.19 vs. 0.33 ± 0.23 mm, respectively, P = 0.006), and more circular (mean eccentricity index 0.08 ± 0.06 vs. 0.10 ± 0.06, stented vs. unstented; P = 0.019). Adherent thrombus was identified in three grafts (all unstented). Our findings highlight the early changes occurring in SVGs after implantation of aorto-coronary bypass conduits, changes that may accelerate vein graft failure. External stenting resulted in a more homogeneous and less eccentric lumen with no thrombus formation.


Neural decoding with kernel-based metric learning.
In studies of the nervous system, the choice of metric for the neural responses is a pivotal assumption. For instance, a well-suited distance metric enables us to gauge the similarity of neural responses to various stimuli and assess the variability of responses to a repeated stimulus-exploratory steps in understanding how the stimuli are encoded neurally. Here we introduce an approach where the metric is tuned for a particular neural decoding task. Neural spike train metrics have been used to quantify the information content carried by the timing of action potentials. While a number of metrics for individual neurons exist, a method to optimally combine single-neuron metrics into multineuron, or population-based, metrics is lacking. We pose the problem of optimizing multineuron metrics and other metrics using centered alignment, a kernel-based dependence measure. The approach is demonstrated on invasively recorded neural data consisting of both spike trains and local field potentials. The experimental paradigm consists of decoding the location of tactile stimulation on the forepaws of anesthetized rats. We show that the optimized metrics highlight the distinguishing dimensions of the neural response, significantly increase the decoding accuracy, and improve nonlinear dimensionality reduction methods for exploratory neural analysis.


Improving predictive models of glaucoma severity by incorporating quality indicators.
In this paper we present an evaluation of the role of reliability indicators in glaucoma severity prediction. In particular, we investigate whether it is possible to extract useful information from tests that would be normally discarded because they are considered unreliable. We set up a predictive modelling framework to predict glaucoma severity from visual field (VF) tests sensitivities in different reliability scenarios. Three quality indicators were considered in this study: false positives rate, false negatives rate and fixation losses. Glaucoma severity was evaluated by considering a 3-levels version of the Advanced Glaucoma Intervention Study scoring metric. A bootstrapping and class balancing technique was designed to overcome problems related to small sample size and unbalanced classes. As a classification model we selected Naïve Bayes. We also evaluated Bayesian networks to understand the relationships between the different anatomical sectors on the VF map. The methods were tested on a data set of 28,778 VF tests collected at Moorfields Eye Hospital between 1986 and 2010. Applying Friedman test followed by the post hoc Tukey's honestly significant difference test, we observed that the classifiers trained on any kind of test, regardless of its reliability, showed comparable performance with respect to the classifier trained only considering totally reliable tests (p-value>0.01). Moreover, we showed that different quality indicators gave different effects on prediction results. Training classifiers using tests that exceeded the fixation losses threshold did not have a deteriorating impact on classification results (p-value>0.01). On the contrary, using only tests that fail to comply with the constraint on false negatives significantly decreased the accuracy of the results (p-value<0.01). Meaningful patterns related to glaucoma evolution were also extracted. Results showed that classification modelling is not negatively affected by the inclusion of less reliable tests in the training process. This means that less reliable tests do not subtract useful information from a model trained using only completely reliable data. Future work will be devoted to exploring new quantitative thresholds to ensure high quality testing and low re-test rates. This could assist doctors in tuning patient follow-up and therapeutic plans, possibly slowing down disease progression.


Muscle anatomy and dynamic muscle function in osteogenesis imperfecta type I.
Results of previous studies suggested that children and adolescents with osteogenesis imperfecta (OI) type I have a muscle force deficit. However, muscle function has only been assessed by static isometric force tests and not in more natural conditions such as dynamic force and power tests. The purpose of this study was to assess lower extremity dynamic muscle function and muscle anatomy in OI type I. The study was performed in the outpatient department of a pediatric orthopedic hospital. A total of 54 individuals with OI type I (6-21 years; 20 male) and 54 age- and sex-matched controls took part in this study. Calf muscle cross-sectional area and density were measured by peripheral quantitative computed tomography. Lower extremity muscle function (peak force per body weight and peak power per body mass) was measured by jumping mechanography through 5 tests: multiple two-legged hopping, multiple one-legged hopping, single two-legged jump, chair-rise test, and heel-rise test. Compared with age- and sex-matched controls, patients with OI type I had smaller muscle size (P = .04) but normal muscle density (P = .21). They also had lower average peak force and lower specific force (peak force/muscle cross-sectional area; all P < .008). Average peak power was lower in patients with OI type I but not significantly so (all P > .054). Children and adolescents with OI type I have, on average, a significant force deficit in the lower limb as measured by dynamic force tests. Nonetheless, these data also show that OI type I is compatible with normal muscle performance in some individuals.


Monitoring during peripheral nerve blockade.
Objective monitors to warn against adverse events specific to the performance of nerve blockade are relatively unsophisticated compared with monitors used during general anesthesia. The purpose of this review is to outline the various monitors available, and how they can be used to reduce nerve block-related complications. Evidence is accumulating for three objective means of preventing complications. Electrical nerve stimulation is insensitive, but specific, at detecting intraneural needle placement, such that if a motor response is obtained with a current of 0.2 mA or less, the needle tip is positioned intraneurally. Ultrasonography allows for reduction of the volume of local anesthetic required to accomplish a nerve block, reducing the potential for systemic toxicity. The value of ultrasound in reducing inadvertent puncture of neighboring structures, while intuitive, is less clear. Finally, injection pressure monitoring helps objectively distinguish between an injection into low versus high compliance tissue (e.g. intrafascicular versus extrafascicular). Peripheral nerve blockade has evolved from an entirely blind procedure to a more reproducible subspecialty. Several monitors exist to objectively guide and monitor needle placement and application of local anesthetics. Because these monitors are complementary, best practice may involve combining all of these monitoring options when performing peripheral nerve blocks.


Yeast genomic expression patterns in response to low-shear modeled microgravity.
The low-shear microgravity environment, modeled by rotating suspension culture bioreactors called high aspect ratio vessels (HARVs), allows investigation in ground-based studies of the effects of microgravity on eukaryotic cells and provides insights into the impact of space flight on cellular physiology. We have previously demonstrated that low-shear modeled microgravity (LSMMG) causes significant phenotypic changes of a select group of Saccharomyces cerevisiae genes associated with the establishment of cell polarity, bipolar budding, and cell separation. However, the mechanisms cells utilize to sense and respond to microgravity and the fundamental gene expression changes that occur are largely unknown. In this study, we examined the global transcriptional response of yeast cells grown under LSMMG conditions using DNA microarray analysis in order to determine if exposure to LSMMG results in changes in gene expression. LSMMG differentially changed the expression of a significant number of genes (1372) when yeast cells were cultured for either five generations or twenty-five generations in HARVs, as compared to cells grown under identical conditions in normal gravity. We identified genes in cell wall integrity signaling pathways containing MAP kinase cascades that may provide clues to novel physiological responses of eukaryotic cells to the external stress of a low-shear modeled microgravity environment. A comparison of the microgravity response to other environmental stress response (ESR) genes showed that 26% of the genes that respond significantly to LSMMG are involved in a general environmental stress response, while 74% of the genes may represent a unique transcriptional response to microgravity. In addition, we found changes in genes involved in budding, cell polarity establishment, and cell separation that validate our hypothesis that phenotypic changes observed in cells grown in microgravity are reflected in genome-wide changes. This study documents a considerable response to yeast cell growth in low-shear modeled microgravity that is evident, at least in part, by changes in gene expression. Notably, we identified genes that are involved in cell signaling pathways that allow cells to detect environmental changes, to respond within the cell, and to change accordingly, as well as genes of unknown function that may have a unique transcriptional response to microgravity. We also uncovered significant changes in the expression of many genes involved in cell polarization and bud formation that correlate well with the phenotypic effects observed in yeast cells when grown under similar conditions. These results are noteworthy as they have implications for human space flight.


First record of Lyme disease Borrelia in the Arctic.
The epidemiology and ecology of Lyme disease is very complex, and its reported geographical distribution is constantly increasing. Furthermore, the involvement of birds in long distance dispersal and their role as reservoir hosts is now well established. In this study, we have shown that sea birds in the Arctic region of Norway carry Ixodes uriae ticks infected with Lyme disease Borrelia garinii spirochetes. Interestingly, DNA sequencing showed that these isolates are closely related to B. garinii previously isolated from birds, as well as from clinical specimens in northern Europe.


Immunochemical characterization of two thyroid-stimulating hormone beta-subunit epitopes.
The epitopes of human thyroid-stimulating hormone (hTSH) recognized by two murine monoclonal antibodies (MAbs), designated MAb 279 and MAb 299, have been characterized. These MAbs are highly specific for the beta-subunit of TSH. The epitope recognized by MAb 279 appears to be completely conserved between bovine and human TSH and partially conserved in the porcine species. The TSH beta-subunit epitope recognized by MAb 299 is only partially conserved between the human, bovine and porcine species. Both MAbs are capable of inhibiting the binding of TSH to its receptor in a TSH radioreceptor assay, indicating that the epitopes either coincide or are located close to the TSH beta-subunit receptor-binding sites. The carbohydrate moieties of the TSH beta-subunit appear to play little or no role in the epitope recognition by MAb 279 or MAb 299 while the integrity of the disulphide bonds are essential. The epitopic recognition may also involve lysine residues, as determined by the immunoreactivity with both MAbs following citraconylation of TSH. In addition, the amino acid sequence region between residues bTSH beta 34-44 could be excised by trypsin digestion of bovine TSH beta (bTSH beta) without eliminating epitopic recognition by either MAb. These results provide further insight into the relationship between the structure of the TSH beta-subunit epitopes and location of the receptor-binding sites.


Carrier screening for cystic fibrosis, Gaucher disease, and Tay-Sachs disease in the Ashkenazi Jewish population: the first 1000 cases at New York University Medical Center, New York, NY.
By late 1993, the genes for cystic fibrosis and Gaucher disease and the mutations common among Ashkenazi Jews had been identified. In response to these advances, heterozygote screening for cystic fibrosis and Gaucher disease was added to the more than 20-year-old Tay-Sachs disease screening program at New York University Medical Center, New York, NY. To review the outcomes from the first 1000 patients screened through this program. Patients and their referring physicians were informed about the new carrier tests. At the time of screening, patients could choose their tests (hexosaminidase A by enzyme analysis for Tay-Sachs disease or mutation analysis for cystic fibrosis and Gaucher disease). All partners of Tay-Sachs and cystic fibrosis carriers were tested. Prenatal diagnosis was offered and performed for carrier couples or mixed-marriage couples in whom the Ashkenazi Jewish partner was a carrier of Gaucher disease. Outcomes were measured by: (1) choice of tests, (2) decisions regarding prenatal diagnosis, and (3) phenotypes of children born to patients who underwent screening. The majority of Ashkenazi Jewish patients chose to have testing for all 3 diseases. If they previously underwent screening for Tay-Sachs disease, then they chose to undergo testing for cystic fibrosis and Gaucher disease. All carrier couples for each of these diseases went on to have prenatal testing. All mixed-marriage couples in whom the Jewish partner was found to be a carrier for Gaucher disease chose to have prenatal diagnosis. One fetus was identified as having cystic fibrosis. Since the program was initiated, no Ashkenazi Jewish baby has been born with any of these diseases at New York University Medical Center. New tests can be readily incorporated into established heterozygote screening programs. The Ashkenazi Jewish population described herein tends to choose testing for all conditions for which heterozygote screening is available.


Germinated barley foodstuff prolongs remission in patients with ulcerative colitis.
Germinated barley foodstuff (GBF) is a prebiotic which increases luminal butyrate production by modulating the microfloral distribution. GBF has been shown to reduce both clinical activity and mucosal damage in active ulcerative colitis (UC) with mild to moderate activity. However, the efficacy of GBF in patients with UC during the remission stage is unknown. The aim of this study was to investigate the efficacy of GBF as a maintenance therapy in patients with UC while in remission. Fifty-nine patients with UC in remission according to Rachmilewitz's clinical activity index (CAI) score of </=4 were enrolled and divided into two groups, control (n=37) and GBF (n=22). Patients in the control group were given conventional treatment alone for 12 months, while patients in the GBF group received conventional therapy plus 20 g of GBF daily. The response to treatments was assessed by monitoring the CAI and endoscopic score according to Matts. Significantly better CAI values were seen in the GBF group at 3, 6, and 12 months compared with the values in the control group. The cumulative recurrence rate in the GBF group with steroid tapering treatment was significantly lower compared with the value in the control group. No side effects related to GBF were observed. GBF appeared to be effective and safe as a maintenance therapy to taper steroid dose and prolong remission in patients with UC.


Fast 2-D ultrasound strain imaging: the benefits of using a GPU.
Deformation of tissue can be accurately estimated from radio-frequency ultrasound data using a 2-dimensional normalized cross correlation (NCC)-based algorithm. This procedure, however, is very computationally time-consuming. A major time reduction can be achieved by parallelizing the numerous computations of NCC. In this paper, two approaches for parallelization have been investigated: the OpenMP interface on a multi-CPU system and Compute Unified Device Architecture (CUDA) on a graphics processing unit (GPU). The performance of the OpenMP and GPU approaches were compared with a conventional Matlab implementation of NCC. The OpenMP approach with 8 threads achieved a maximum speed-up factor of 132 on the computing of NCC, whereas the GPU approach on an Nvidia Tesla K20 achieved a maximum speed-up factor of 376. Neither parallelization approach resulted in a significant loss in image quality of the elastograms. Parallelization of the NCC computations using the GPU, therefore, significantly reduces the computation time and increases the frame rate for motion estimation.


Cellular Senescence as the Causal Nexus of Aging.
In this paper we present cellular senescence as the ultimate driver of the aging process, as a "causal nexus" that bridges microscopic subcellular damage with the phenotypic, macroscopic effect of aging. It is important to understand how the various types of subcellular damage correlated with the aging process lead to the larger, visible effects of anatomical aging. While it has always been assumed that subcellular damage (cause) results in macroscopic aging (effect), the bridging link between the two has been hard to define. Here, we propose that this bridge, which we term the "causal nexus", is in fact cellular senescence. The subcellular damage itself does not directly cause the visible signs of aging, but rather, as the damage accumulates and reaches a critical mass, cells cease to proliferate and acquire the deleterious "senescence-associated secretory phenotype" (SASP) which then leads to the macroscopic consequences of tissue breakdown to create the physiologically aged phenotype. Thus senescence is a precondition for anatomical aging, and this explains why aging is a gradual process that remains largely invisible during most of its progression. The subcellular damage includes shortening of telomeres, damage to mitochondria, aneuploidy, and DNA double-strand breaks triggered by various genetic, epigenetic, and environmental factors. Damage pathways acting in isolation or in concert converge at the causal nexus of cellular senescence. In each species some types of damage can be more causative than in others and operate at a variable pace; for example, telomere erosion appears to be a primary cause in human cells, whereas activation of tumor suppressor genes is more causative in rodents. Such species-specific mechanisms indicate that despite different initial causes, most of aging is traced to a single convergent causal nexus: senescence. The exception is in some invertebrate species that escape senescence, and in non-dividing cells such as neurons, where senescence still occurs, but results in the SASP rather than loss of proliferation plus SASP. Aging currently remains an inevitable endpoint for most biological organisms, but the field of cellular senescence is primed for a renaissance and as our understanding of aging is refined, strategies capable of decelerating the aging process will emerge.


Phenolic-Rich Baccaurea angulata Modulates Inflammatory Biomarkers of Atherosclerosis.
Cardiovascular disease (CVD) is the leading and the most critical type of chronic disease. Atherosclerosis is the most common cause of CVD. Inflammation has been progressively acknowledged as a vital and central player in the pathophysiology of atherosclerosis. Baccaurea angulata is an underutilized fruit of the island of Borneo. It was obtained from Bau, Sarawak, Malaysia. In our previous studies, B. angulata did not only increase antioxidant enzyme activities, but also slowed the lipid peroxidation process in high-cholesterol-fed rabbits. It was hypothesized that B. angulata fruit would exert an anti-inflammatory effect. This study, therefore, aimed at evaluating and comparing the effects of three different B. angulata whole fruit (WF) juice doses on 11 serum inflammatory biomarkers of atherosclerosis. Thirty-five male New Zealand white rabbits were divided into seven groups (n=5). Group CH was fed 1% cholesterol diet only, group C1 was fed 1% cholesterol diet and 0.5 ml/kg/day B. angulata WF juice, group C2 was fed 1% cholesterol diet and 1.0 ml/kg/day B. angulata WF juice, group C3 was fed 1% cholesterol diet and 1.5 ml/kg/day B. angulata WF juice, group N was fed standard pellet only, group N1 was fed standard pellet and 0.5 ml/kg/day B. angulata WF juice, and group N2 was fed standard pellet and 1.0 ml/kg/day B. angulata WF juice for 12 weeks. The administration of the various juices reduced the concentrations of induced serum inflammatory biomarkers. This protective effect of B. angulata fruit against cardiovascular risk might be due to its polyphenol content.


Molecular Characterization of Methicillin Resistant Staphylococcus aureus in West Bank-Palestine.
Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a public health threat and a major cause of hospital-acquired and community-acquired infections. This study aimed to investigate the genetic diversity of MRSA isolates from 2015 to 2017 and to characterize the major MRSA clones and anti-biogram trends in Palestine. Methodology: Isolates were obtained from 112 patients admitted to different hospitals of West Bank and East Jerusalem, originating from different clinical sources. Antibiotic susceptibility patterns, staphylococcal chromosomal cassette mec (SCCmec) typing, and Staphylococcus aureus protein A (spa) typing were determined. Also, a panel of toxin genes and virulence factors was studied, including: Panton-Valentine Leukocidin (PVL), ACME-arcA, Toxic Shock Syndrome Toxin-1 (TSST-1), and Exfoliative Toxin A (ETA). Results: Of the 112 confirmed MRSA isolates, 100% were resistant to all β-lactam antibiotics. Resistance rates to other non- β-lactam classes were as the following: 18.8% were resistant to trimethoprim-sulfamethoxazole, 23.2% were resistant to gentamicin, 34.8% to clindamycin, 39.3% to ciprofloxacin, and 63.4% to erythromycin. All MRSA isolates were susceptible to vancomycin (100%). Of all isolates, 32 isolates (28.6%) were multidrug- resistant (MDR). The majority of the isolates were identified as SCCmec type IV (86.6%). The molecular typing identified 29 spa types representing 12 MLST-clonal complexes (CC). The most prevalent spa types were: spa type t386 (CC1)/(12.5%), spa type t044 (CC80)/(10.7%), spa type t008 (CC8)/(10.7%), and spa type t223 (CC22)/(9.8%). PVL toxin gene was detected in (29.5%) of all isolates, while ACME-arcA gene was present in 18.8% of all isolates and 23.2% had the TSST-1 gene. The two most common spa types among the TSST-1positive isolates were the spa type t223 (CC22)/(Gaza clone) and the spa type t021 (CC30)/(South West Pacific clone). All isolates with the spa type t991 were ETA positive (5.4%). USA-300 clone (spa type t008, positive for PVL toxin gene and ACME-arcA genes) was found in nine isolates (8.0%). Conclusions: Our results provide insights into the epidemiology of MRSA strains in Palestine. We report a high diversity of MRSA strains among hospitals in Palestine, with frequent SCCmec type IV carriage. The four prominent clones detected were: t386-IV/ CC1, the European clone (t044/CC80), Gaza clone (t223/CC22), and the USA-300 clone (t008/CC8).


Nova2 interacts with a cis-acting polymorphism to influence the proportions of drug-responsive splice variants of SCN1A.
An intronic polymorphism in the SCN1A gene, which encodes a neuronal sodium-channel alpha subunit, has been previously associated with the dosing of two commonly used antiepileptic drugs that elicit their pharmacologic action primarily at this ion-channel subunit. This study sought to characterize the functional effects of this polymorphism on alternative splicing of SCN1A and to explore the potential for modulating the drug response in the pharmacologically unfavorable genotype by identification of a splice modifier acting on SCN1A. The effects of the genotype at the SCN1A IVS5N+5 G-->A polymorphism on SCN1A splice-variant proportions and the consequences of increased expression of splice modifiers were investigated both in human temporal neocortex tissue and in a cellular minigene expression system. Quantitative real-time polymerase chain reaction was used to quantify the amounts of SCN1A transcripts forms. We show that the polymorphism has a dramatic effect on the proportions of neonate and adult alternative transcripts of SCN1A in adult brain tissue and that the effect of the polymorphism also appears to be modified by Nova2 expression levels. A minigene expression system confirms both the effect of the polymorphism on transcript proportions and the role of Nova2 in the regulation of splicing, with higher Nova2 expression increasing the proportion of the neonate form. A larger Nova2-mediated effect was detected in the AA genotype that is associated with increased dose requirements. The effects of Nova2 on modulation of the alternative splicing of 17 other neuronally expressed genes were investigated, and no effect was observed. These findings emphasize the emerging role of genetic polymorphisms in modulation of drug effect and illustrate both alternative splicing as a potential therapeutic target and the importance of considering the activity of compounds at alternative splice forms of drug targets in screening programs.


Evolutionary transitions and artificial life.
A major challenge for artificial life is to synthesize the evolutionary transitions that have repeatedly formed differentiated higher-level entities from cooperative organizations of lower-level entities, producing the nested hierarchical structure of living processes. This article identifies the key elements and relationships that must be incorporated or synthesized in an artificial life system if these transitions are to emerge. The processes currently included in artificial life systems are unable to provide an adequate basis for the emergence of the complex cooperative organization that is essential to the transitions. A new theory of the evolution of cooperative organization is developed that points to the additional processes that must be included in artificial life systems to underpin the emergence of the transitions.


Ambident reactivities of pyridone anions.
The kinetics of the reactions of the ambident 2- and 4-pyridone anions with benzhydrylium ions (diarylcarbenium ions) and structurally related Michael acceptors have been studied in DMSO, CH(3)CN, and water. No significant changes of the rate constants were found when the counterion was varied (Li(+), K(+), NBu(4)(+)) or the solvent was changed from DMSO to CH(3)CN, whereas a large decrease of nucleophilicity was observed in aqueous solution. The second-order rate constants (log k(2)) correlated linearly with the electrophilicity parameters E of the electrophiles according to the correlation log k(2) = s(N + E) (Angew. Chem., Int. Ed. Engl. 1994, 33, 938-957), allowing us to determine the nucleophilicity parameters N and s for the pyridone anions. The reactions of the 2- and 4-pyridone anions with stabilized amino-substituted benzhydrylium ions and Michael acceptors are reversible and yield the thermodynamically more stable N-substituted pyridones exclusively. In contrast, highly reactive benzhydrylium ions (4,4'-dimethylbenzhydrylium ion), which react with diffusion control, give mixtures arising from N- and O-attack with the 2-pyridone anion and only O-substituted products with the 4-pyridone anion. For some reactions, rate and equilibrium constants were determined in DMSO, which showed that the 2-pyridone anion is a 2-4 times stronger nucleophile, but a 100 times stronger Lewis base than the 4-pyridone anion. Quantum chemical calculations at MP2/6-311+G(2d,p) level of theory showed that N-attack is thermodynamically favored over O-attack, but the attack at oxygen is intrinsically favored. Marcus theory was employed to develop a consistent scheme which rationalizes the manifold of regioselectivities previously reported for the reactions of these anions with electrophiles. In particular, Kornblum's rationalization of the silver ion effect, one of the main pillars of the hard and soft acid/base concept of ambident reactivity, has been revised. Ag(+) does not reverse the regioselectivity of the attack at the 2-pyridone anion by increasing the positive charge of the electrophile but by blocking the nitrogen atom of the 2-pyridone anion.


Near-infrared fluorescence contrast-enhanced imaging with area illumination and area detection: the forward imaging problem.
Fluorescence frequency-domain photon migration measurements were acquired from tissue phantoms, each containing a fluorescent target, by means of area illumination and area detection on the same surface and for the first time, to our knowledge, compared with predictions computed with a numerical solution to the coupled photon diffusion equations. We accomplished area illumination and area detection using a planar, intensity-modulated excitation light source and a gain-modulated intensified charge-coupled device camera, respectively. A 1-ml vessel containing 1-microm solution of Indocyanine Green in 1% Liposyn was immersed 1 cm deep in each 512-ml tissue phantom. For most tissue phantoms, the background surrounding the 1-ml target was composed of Liposyn solution containing Indocyanine Green or 3,3'-Diethylthiatricarbocyanine Iodide such that the target-to-background ratio of fluorescence yield was > or = 10:1. Measurements of fluorescence modulation amplitude and phase were predicted with a mean error ranging from 10.1% to 13.6% and 0.56 degrees to 1.72 degrees, respectively. These numbers are similar to those obtained by use of single-pixel frequency-domain photon migration techniques and validate the potential use of area illumination and area detection for biomedical imaging of tissues. Results also demonstrate that target-to-background ratios of fluorescence yield and fluorescence lifetime significantly affect target detectability.


Heterogeneities in glaciofluvial deposits using an example from new hampshire.
The strong influence of subsurface heterogeneity on contaminant migration and in situ remediation calls for an improved understanding of its origins and more efficient methods of characterization. Accordingly, an outcrop study of physical and chemical heterogeneity was conducted in a glaciofluvial deposit in Deerfield, New Hampshire, in order to uncover processes controlling the spatial variation of sediment properties and evaluate the extent to which geologic information can be used to characterize the observed variation. The results indicate that physical and chemical properties at the Deerfield site have distinctly different spatial correlation structures. Lithologic facies explain 31% to 60% of the variation in permeability, dithionite citrate (DC)-extractable manganese, and DC-extractable aluminum. Lithofacies bounding surfaces do not separate regions of significantly different DC-extractable iron; instead, 49% of its variation is explained by sediment color. Color also accounts for 34% of the variation in DC-extractable aluminum and 60% of the variation in DC-extractable manganese. Strong relationships with sediment facies and/or color enable detailed mapping of permeability, extractable iron, and extractable manganese. Differences in the geometries of iron and manganese enrichment, petrographic observations, and scanning electron microscope analyses indicate that (hydr)oxide grain coatings originated from the postdepositional weathering of biotite and garnet, coupled with local, redox-driven redistribution of the liberated iron and manganese. The findings suggest that lithofacies and color information can aid the characterization and modeling of heterogeneity at similar carbon-poor glaciofluvial sites.


Clinical, dermoscopic and histopathological features of eccrine poroid neoplasms.
Eccrine poroma (EP) belongs to the group of benign adnexal tumors, but cases of eccrine porocarcinoma (EPC) arising on long-standing and untreated EP suggest a certain risk of malignant transformation. To describe the clinical, dermoscopic and histopathologic features associated with different extremes in the spectrum of eccrine poroid neoplasms and to review the according literature. A retrospective analysis of patient characteristics and morphologic features associated with EP, EPC in situ and invasive porocarcinoma patients who attended two skin lesion clinics in Italy and Turkey between 2010 and 2011. A total of 4 cases including 1 EP, 1 EPC in situ and 2 cases of invasive EPC in 4 patients were analyzed. Recent changes including bleeding, ulceration and sudden enlargement of a pre-existing lesion were associated with malignant transformation. Dermoscopically, polymorphous vessels consisting of coiled, hairpin and linear vessels were seen at all stages of progression. Histopathological findings of EPC in situ were limited to the epidermal component and were evident only at higher magnification. Clinicians and pathologists should carefully evaluate EPs with a recent history of change and erosion in order to avoid overlooking the potential development of EPC.