Micromechanics of ultra-toughened electrospun PMMA/PEO fibres as revealed by in-situ tensile testing in an electron microscope.
A missing cornerstone in the development of tough micro/nano fibre systems is an understanding of the fibre failure mechanisms, which stems from the limitation in observing the fracture of objects with dimensions one hundredth of the width of a hair strand. Tensile testing in the electron microscope is herein adopted to reveal the fracture behaviour of a novel type of toughened electrospun poly(methyl methacrylate)/poly(ethylene oxide) fibre mats for biomedical applications. These fibres showed a toughness more than two orders of magnitude greater than that of pristine PMMA fibres. The in-situ microscopy revealed that the toughness were not only dependent on the initial molecular alignment after spinning, but also on the polymer formulation that could promote further molecular orientation during the formation of micro/nano-necking. The true fibre strength was greater than 150 MPa, which was considerably higher than that of the unmodified PMMA (17 MPa). This necking phenomenon was prohibited by high aspect ratio cellulose nanocrystal fillers in the ultra-tough fibres, leading to a decrease in toughness by more than one order of magnitude. The reported necking mechanism may have broad implications also within more traditional melt-spinning research.


Evidence of two-state reactivity in alkane hydroxylation by Lewis-acid bound copper-nitrene complexes.
The behavior of the Lewis-acid adducts of two copper-nitrene [Cu(NR)](+) complexes in nitrene-transfer and H-atom abstraction reactions have been demonstrated to depend on the nature of the nitrene substituents. Two-state reactivity, in which a singlet ground state and a nearby triplet excited-state both contribute, provides a useful model for interpreting reactivity trends of the two compounds.


Standard work tools for dynamic stereoelectroencephalography using ROSA: naming convention and perioperative planning.
The grid-based orthogonal placement of depth electrodes (DEs), initially defined by Jean Talairach and Jean Bancaud, is known as stereo-electroencephalography (sEEG). Although acceptance in the United States was initially slow, advances in imaging and technology have spawned a proliferation of North American epilepsy centers offering sEEG. Despite publications highlighting minimal access techniques and varied indications, standard work for phase I targeted DE has not been defined. In this article, the authors propose the term "dynamic sEEG" and define standard work tools and related common data elements to promote uniformity in the field. A multidisciplinary approach from July to August 2016 resulted in the production of 4 standard work tools for dynamic sEEG using ROSA: 1) a 34-page illustrated manual depicting a detailed workflow; 2) a planning form to collocate all the phase I data; 3) a naming convention for DEs that encodes the data defining it; and 4) a reusable portable perioperative planning and documentation board. A retrospective review of sEEG case efficiency was performed comparing those using standard work tools (between July 2016 and April 2017) with historical controls (between March 2015 and June 2016). The standard work tools were then instituted at another epilepsy surgery center, and the results were recorded. The process for dynamic sEEG was formally reviewed, including anesthesia, positioning, perioperative nursing guidelines, surgical steps, and postoperative care for the workflow using cranial fixation and ROSA-guided placement. There was a 40% improvement in time per electrode, from 44.7 ± 9.0 minutes to 26.9 ± 6.5 minutes (p = 0.0007) following the development and use of the manual, the naming convention, and the reusable portable perioperative planning and documentation board. This standardized protocol was implemented at another institution and yielded a time per electrode of 22.3 ± 4.4 minutes. The authors propose the term dynamic sEEG for stereotactic depth electrodes placed according to phase I workup data with the intention of converting to ablation. This workflow efficiency can be optimized using the standard work tools presented. The authors also propose a novel naming convention that encodes critical data and allows portability among providers. Use of a planning form for common data elements optimizes research, and global adoption could facilitate multicenter studies correlating phase I modality and seizure onset zone identification.


Diverse Borrelia burgdorferi strains in a bird-tick cryptic cycle.
The blacklegged tick Ixodes scapularis is the primary vector of the most prevalent vector-borne zoonosis in North America, Lyme disease (LD). Enzootic maintenance of the pathogen Borrelia burgdorferi by I. scapularis and small mammals is well documented, whereas its "cryptic" maintenance by other specialist ticks and wildlife hosts remains largely unexplored because these ticks rarely bite humans. We quantified B. burgdorferi infection in a cryptic bird-rabbit-tick cycle. Furthermore, we explored the role of birds in maintaining and moving B. burgdorferi strains by comparing their genetic diversity in this cryptic cycle to that found in cycles vectored by I. scapularis. We examined birds, rabbits, and small mammals for ticks and infection over a 4-year period at a focal site in Michigan, 90 km east of a zone of I. scapularis invasion. We mist netted 19,631 birds that yielded 12,301 ticks, of which 86% were I. dentatus, a bird-rabbit specialist. No resident wildlife harbored I. scapularis, and yet 3.5% of bird-derived ticks, 3.6% of rabbit-derived ticks, and 20% of rabbit ear biopsy specimens were infected with B. burgdorferi. We identified 25 closely related B. burgdorferi strains using an rRNA gene intergenic spacer marker, the majority (68%) of which had not been reported previously. The presence of strains common to both cryptic and endemic cycles strongly implies bird-mediated dispersal. Given continued large-scale expansion of I. scapularis populations, we predict that its invasion into zones of cryptic transmission will allow for bridging of novel pathogen strains to humans and animals.


Pharmacology of Ranolazine versus Common Cardiovascular Drugs in Patients with Early Diastolic Dysfunction Induced by Anthracyclines or Nonanthracycline Chemotherapeutics: A Phase 2b Minitrial.
We have reported that anthracyclines and nonanthracycline chemotherapeutics caused diastolic dysfunction in cancer patients without cardiovascular risk factors. Diastolic dysfunction occurred as early as 1 week after the last chemotherapy cycle and manifested as impaired myocardial relaxation at echocardiography or persistent elevations of B-type natriuretic peptide (BNP) or troponin. The antianginal drug ranolazine shows cardiac relaxant effects that we considered of value to treat early diastolic dysfunction induced by cancer drugs; therefore, 24 low-risk patients with post-chemotherapy diastolic dysfunction were randomized (1:1) to ranolazine or the investigator's choice of common cardiovascular drugs, such as β-blockers and/or angiotensin-converting enzyme inhibitors or loop diuretics (best standard therapy, BST). After 5 weeks, 12 of 12 patients on ranolazine recovered from diastolic dysfunction, whereas 3 of 12 patients on BST did not improve; however, adverse events (not serious) were apparently more frequent for ranolazine than for BST (4/12 vs. 1/12). Ranolazine did not lower blood pressure, whereas BST reduced systolic pressure and caused a trend toward a reduced diastolic pressure. Most patients at randomization showed tachycardia resulting from chemotherapy-related anemia. Hemoglobin recovery contributed to normalizing heart rate in these patients; however, some patients in the ranolazine arm developed tachycardia through chronotropic effects of high BNP levels and returned to a normal heart rate through the effects of ranolazine on decreasing BNP levels. This minitrial describes the potential effects of ranolazine on relieving chemotherapy-related diastolic dysfunction; however, clinical implications of these findings need to be characterized by studies with an adequate sample size. SIGNIFICANCE STATEMENT: The antianginal drug ranolazine causes cardiac relaxant effects that might relieve diastolic dysfunction. In a clinical pharmacology study, 24 patients were randomized (1:1) to receive ranolazine or common cardiovascular drugs to treat early diastolic dysfunction induced by anthracycline-based or nonanthracycline chemotherapy. Ranolazine relieved diastolic dysfunction in these patients. The safety profile of ranolazine in cancer patients is similar to that of the general population. Compared with common cardiovascular drugs, ranolazine relieved diastolic dysfunction without lowering blood pressure. The sample size of this study was nonetheless too small to permit considerations about the potential clinical value of ranolazine for oncologic patients with early diastolic dysfunction induced by anthracyclines or nonanthracycline chemotherapeutics. This information should be obtained by studies with an adequate sample size.


Hepatic microvascular dysfunction in endotoxemic rats after acute ethanol administration.
A high concentration of ethanol is reported to cause hepatic microvascular dysfunction. However, little is known about the effect of ethanol on hepatic microcirculation in endotoxemic animals. The objective of this study was to determine whether endotoxemia enhances the hepatic microvascular dysfunction induced by acute ethanol administration. Intravital videomicroscopy was used to monitor leukocyte recruitment, number of nonperfused sinusoids, and flow velocity of erythrocytes (RBC) labeled with fluorescein isothiocyanate (FITC) in the livers of male Wistar rats that were administered ethanol (20%, 3 g/kg; or 40%, 6 g/kg) from a gastric tube. Flow velocity of RBC in sinusoids was measured with an off-line velocimeter. Plasma tumor necrosis factor (TNF)-alpha levels were also measured. In some experiments, rats were injected with 2 mg/kg of lipopolysaccharides (LPS) intraperitoneally at 16 hr before the experiments, and the same protocol was performed. Although FITC-RBC velocity was initially increased by both 20% and 40% ethanol in control rats, it was reduced by only 40% ethanol at 60 min. In LPS-treated rats, the FITC-RBC velocity also was increased initially but was reduced at 60 and 30 min by 20% and 40% ethanol, respectively. Only 40% ethanol caused leukostasis in the pericentral region of control rats. In LPS-treated rats, however, leukostasis was noted in the midzonal and pericentral regions of liver after both 20% and 40% ethanol administration. Ethanol increased plasma TNF-alpha levels only in LPS-treated rats. These results suggest that LPS synergistically enhances ethanol-induced hepatic microvascular dysfunction and liver injury, especially in the midzonal region via coagulation, which may be mediated by TNF-alpha.


Homozygosity and linkage-disequilibrium mapping of the urofacial (Ochoa) syndrome gene to a 1-cM interval on chromosome 10q23-q24.
The urofacial (Ochoa) syndrome (UFS) is a rare autosomal recessive disease characterized by congenital obstructive uropathy and abnormal facial expression. The patients present with enuresis, urinary-tract infection, hydronephrosis, and voiding dysfunctions as a result of neurogenic bladders. To map the UFS gene, a genome screen using a combination of homozygosity-mapping and DNA-pooling strategies was performed in 20 selected patients, one patient pool, and three control pools (unaffected relatives). After analyses of 36 randomly chosen markers, D10S677 was identified as being linked to and associated with UFS, as suggested by a significant excess of homozygosity in patients compared with that in unaffected relatives (P < 10(-6)), as well as by the allelic-frequency differences between the patient pool and control pools. Ten additional markers flanking D10S677 and covering a 22-cM region then were analyzed to fine-map the UFS gene by use of haplotype (linkage disequilibrium) analysis. All 31 patients were found to be homozygous for two closely linked markers (D10S1726 and D10S198) located approximately 5 cM telomeric to D10S677, whereas only 12% of the unaffected relatives were homozygous for both markers (P < 10(-19)). Several patients are heterozygous at two markers immediately flanking D10S1726/D10S198, one on the centromeric side (D10S1433) and the other on the telomeric side (D10S603). These recombinational events place the UFS gene near D10S1726/D10S198 and within a 1-cM interval defined by D10S1433 and D10S603 on chromosome 10q23-q24.


Evaluation of the placement of mobile fruit and vegetable vendors to alleviate food deserts in New York City.
In 2008, the New York City (NYC) health department licensed special mobile produce vendors (Green Carts) to increase access to fruits and vegetables in neighborhoods with the lowest reported fruit and vegetable consumption and the highest obesity rates. Because economic incentives may push vendors to locate in more trafficked, less produce-deprived areas, we examined characteristics of areas with and without Green Carts to explore whether Carts are positioned to reach the intended populations. Using ArcGIS software, we mapped known NYC Green Cart locations noted through 2013 and generated a list of potential (candidate) sites where Carts could have located. We compared the food environment (via categorizing "healthy" or "unhealthy" food stores using federal classification codes corroborated by online storefront images) and other factors that might explain Cart location (eg, demographic, business, neighborhood characteristics) near actual and candidate sites descriptively and inferentially. Seven percent of Green Carts (n = 265) were in food deserts (no healthy stores within one-quarter mile) compared with 36% of candidate sites (n = 644, P < .001). Most Carts (78%) were near 2 or more healthy stores. Green Carts had nearly 60 times the odds of locating near subway stops (P < .001), were closer to large employers (odds ratio [OR], 6.4; P < .001), other food stores (OR, 14.1; P < .001), and in more populous tracts (OR, 2.9, P <.01) compared with candidate sites. Green Carts were rarely in food deserts and usually had multiple healthy stores nearby, suggesting that Carts may not be serving the neediest neighborhoods. Exploration of Carts' benefits in non-food desert areas is needed, but incentivizing vendors to locate in still-deprived places may increase program impact.


Percutaneous retrogasserian glycerol rhizolysis in the management of trigeminal neuralgia.
Sixty patients with trigeminal neuralgia or atypical facial pain were followed for an average of 1 year after percutaneous retrogasserian glycerol rhizolysis. The procedure was initially effective in relieving pain in 80% of the patients with typical trigeminal neuralgia and symptomatic trigeminal neuralgia secondary to multiple sclerosis. However, life-table analysis indicated that 50% of this group had persistence or recurrence of pain within 18 months after the operation. Percutaneous retrogasserian glycerol rhizolysis was ineffective in relieving atypical trigeminal neuralgia or atypical facial pain. Minor complications occurred in 23% of patients, and major morbidity was seen in 1.6%. Facial sensory loss which persisted for more than 1 month was found in 72% of patients, corneal hypesthesia occurred in 15%, and an additional 7% had corneal anesthesia. The data indicate that the success of percutaneous retrogasserian glycerol rhizolysis in relieving trigeminal neuralgia is directly related to the production of facial sensory loss.


Homosalate aggravates the invasion of human trophoblast cells as well as regulates intracellular signaling pathways including PI3K/AKT and MAPK pathways.
Homosalate is an organic ultraviolet filter used in most sunscreens but has been reported to be toxic to marine organisms. The estrogenic activity of homosalate has also been reported, but its endocrine-disrupting effect remains unclear. Although homosalate has been detected in human placental tissues, its effect on the survival of human trophoblast cells needs to be investigated. Therefore, in this study, we evaluated if HTR8/SVneo, a human trophoblast cell line, treated with homosalate showed decreasing proliferative activity in a dose-dependent manner. Homosalate promoted the death of HTR8/SVneo cells with elevated lipid peroxidation and intracellular Ca2+ concentration. It also induced endoplasmic reticulum stress and mitochondrial morphological disturbances associated with the differentiation of human trophoblast cells. However, when the intracellular Ca2+ or reactive oxygen species were removed using BAPTA-AM or N-acetyl-L-cysteine (NAC), the cell proliferation suppressed by homosalate was restored. Homosalate also significantly inhibited the invasion of HTR8/SVneo cells. Furthermore, it modulated phosphoinositide 3-kinase (PI3K)/AKT and mitogen-activated protein kinase (MAPK) signaling pathways, which were involved in the cross-talk between both signaling pathways in HTR8/SVneo cells. Thus, homosalate adversely affects the survival, proliferation, and invasiveness of human trophoblast cells and therefore pregnant women should practice caution while using personal care products containing homosalate.


Are Leukaemic Stem Cells Restricted to a Single Cell Lineage?
Cancer-stem-cell theory states that most, if not all, cancers arise from a stem/uncommitted cell. This theory revolutionised our view to reflect that cancer consists of a hierarchy of cells that mimic normal cell development. Elegant studies of twins who both developed acute lymphoblastic leukaemia in childhood revealed that at least two genomic insults are required for cancer to develop. These 'hits' do not appear to confer a growth advantage to cancer cells, nor do cancer cells appear to be better equipped to survive than normal cells. Cancer cells created by investigators by introducing specific genomic insults generally belong to one cell lineage. For example, transgenic mice in which the LIM-only 2 (LMO2,associated with human acute T-lymphoblastic leukaemia) and BCR-ABLp210 (associated with human chronic myeloid leukaemia) oncogenes were active solely within the haematopoietic stem-cell compartment developed T-lymphocyte and neutrophil lineage-restricted leukaemia, respectively. This recapitulated the human form of these diseases. This 'hardwiring' of lineage affiliation, either throughout leukaemic stem cell development or at a particular stage, is different to the behaviour of normal haematopoietic stem cells. While normal cells directly commit to a developmental pathway, they also remain versatile and can develop into a terminally differentiated cell that is not part of the initial lineage. Many cancer stem cells do not have this versatility, and this is an essential difference between normal and cancer stem cells. In this report, we review findings that support this notion.


Origin and expansion of the mosquito Aedes aegypti in Madeira Island (Portugal).
Historically known as the yellow fever mosquito, Aedes aegypti invaded Madeira Island in 2005 and was the vector of the island's first dengue outbreak in 2012. We have studied genetic variation at 16 microsatellites and two mitochondrial DNA genes in temporal samples of Madeira Island, in order to assess the origin of the invasion and the population structure of this mosquito vector. Our results indicated at least two independent colonization events occurred on the island, both having a South American source population. In both scenarios, Venezuela was the most probable origin of these introductions, a result that is in accordance with the socioeconomic relations between this country and Madeira Island. Once introduced, Ae. aegypti has rapidly expanded along the southern coast of the island and reached a maximum effective population size (Ne) in 2012, coincident with the dengue epidemic. After the outbreak, there was a 10-fold reduction in Ne estimates, possibly reflecting the impact of community-based vector control measures implemented during the outbreak. These findings have implications for mosquito surveillance not only for Madeira Island, but also for other European regions where Aedes mosquitoes are expanding.


Clock-like mutational processes in human somatic cells.
During the course of a lifetime, somatic cells acquire mutations. Different mutational processes may contribute to the mutations accumulated in a cell, with each imprinting a mutational signature on the cell's genome. Some processes generate mutations throughout life at a constant rate in all individuals, and the number of mutations in a cell attributable to these processes will be proportional to the chronological age of the person. Using mutations from 10,250 cancer genomes across 36 cancer types, we investigated clock-like mutational processes that have been operating in normal human cells. Two mutational signatures show clock-like properties. Both exhibit different mutation rates in different tissues. However, their mutation rates are not correlated, indicating that the underlying processes are subject to different biological influences. For one signature, the rate of cell division may influence its mutation rate. This study provides the first survey of clock-like mutational processes operating in human somatic cells.


Cutpoint selection for categorizing a continuous predictor.
This article presents a new approach for choosing the number of categories and the location of category cutpoints when a continuous exposure variable needs to be categorized to obtain tabular summaries of the exposure effect. The optimum categorization is defined as the partition that minimizes a measure of distance between the true expected value of the outcome for each subject and the estimated average outcome among subjects in the same exposure category. To estimate the optimum partition, an efficient nonparametric estimate of the unknown regression function is substituted into a formula for the asymptotically optimum categorization. This new approach is easy to implement and it outperforms existing cutpoint selection methods.


Processed meat intake and chronic disease morbidity and mortality: An overview of systematic reviews and meta-analyses.
Despite the nutritional value of meat, a large volume of reviews and meta-analyses suggests that processed meat intake is associated with an increased risk of chronic diseases. However, assessments of the quality of these published reviews internal validity are generally lacking. We systematically reviewed and assessed the quality alongside summarizing the results of previously published systematic reviews and meta-analyses that examined the association between processed meat intake and cancers, type II diabetes (T2D), and cardiovascular diseases (CVD). Reviews and meta-analyses published until May 2018 were identified through a systematic literature search in the databases MEDLINE and EMBASE, and reference lists of included reviews. The quality of the systematic reviews and meta-analyses was assessed using A Measurement Tool to Assess Systematic Reviews (AMSTAR). All eligible reviews had to comply with two quality requirements: providing sufficient information on quality assessment of the primary studies and a comprehensive search. The results were summarized for T2D, CVD, and each of the different cancer types. The certainty in the estimates of the individual outcomes was rated using the Grading of Recommendations, Assessment, Development and Evaluations (GRADE) method. In total, 22 systematic reviews were eligible and thus included in this review. More than 100 reviews were excluded because quality assessment of the primary studies had not been performed. The AMSTAR score of the included reviews ranged from 5 to 8 indicating moderate quality. Overall, the quality assessments of primary studies of the reviews are generally lacking; the scientific quality of the systematic reviews reporting positive associations between processed meat intake and risk of various cancers, T2D and CVD is moderate, and the results from case-control studies suggest more often a positive association than the results from cohort studies. The overall certainty in the evidence was very low across all individual outcomes, due to serious risk of bias and imprecision.


A novel approach to non-biased systematic random sampling: a stereologic estimate of Purkinje cells in the human cerebellum.
Non-biased systematic sampling using the principles of stereology provides accurate quantitative estimates of objects within neuroanatomic structures. However, the basic principles of stereology are not optimally suited for counting objects that selectively exist within a limited but complex and convoluted portion of the sample, such as occurs when counting cerebellar Purkinje cells. In an effort to quantify Purkinje cells in association with certain neurodegenerative disorders, we developed a new method for stereologic sampling of the cerebellar cortex, involving calculating the volume of the cerebellar tissues, identifying and isolating the Purkinje cell layer and using this information to extrapolate non-biased systematic sampling data to estimate the total number of Purkinje cells in the tissues. Using this approach, we counted Purkinje cells in the right cerebella of four human male control specimens, aged 41, 67, 70 and 84 years, and estimated the total Purkinje cell number for the four entire cerebella to be 27.03, 19.74, 20.44 and 22.03 million cells, respectively. The precision of the method is seen when comparing the density of the cells within the tissue: 266,274, 173,166, 167,603 and 183,575 cells/cm3, respectively. Prior literature documents Purkinje cell counts ranging from 14.8 to 30.5 million cells. These data demonstrate the accuracy of our approach. Our novel approach, which offers an improvement over previous methodologies, is of value for quantitative work of this nature. This approach could be applied to morphometric studies of other similarly complex tissues as well.


Histone deacetylases as transcriptional activators? Role reversal in inducible gene regulation.
Histone deacetylation enzymes have often been associated with the suppression of eukaryotic gene transcription. In contrast, recent studies of inducible gene regulation indicate that protein deacetylation can also be required as a transcriptional activation signal. The concept of protein deacetylation as a requirement for transcription activation seems to contradict earlier conclusions about the function of deacetylation in gene suppression. However, in the context of a more global interpretation, these opposing effects of deacetylation imply its dynamic role in the overall control of gene expression. The exact requirement for deacetylation differs among promoters, depending on their specific architecture and regulation scenario.


Multiparameter Continuous Physiological Monitoring Technologies in Neonates Among Health Care Providers and Caregivers at a Private Tertiary Hospital in Nairobi, Kenya: Feasibility, Usability, and Acceptability Study.
Continuous physiological monitoring technologies are important for strengthening hospital care for neonates, particularly in resource-constrained settings, and understanding user perspectives is critical for informing medical technology design, development, and optimization. This study aims to assess the feasibility, usability, and acceptability of 2 noninvasive, multiparameter, continuous physiological monitoring technologies for use in neonates in an African health care setting. We assessed 2 investigational technologies from EarlySense and Sibel, compared with the reference Masimo Rad-97 technology through in-depth interviews and direct observations. A purposive sample of health care administrators, health care providers, and caregivers at Aga Khan University Hospital, a tertiary, private hospital in Nairobi, Kenya, were included. Data were analyzed using a thematic approach in NVivo 12 software. Between July and August 2020, we interviewed 12 health care providers, 5 health care administrators, and 10 caregivers and observed the monitoring of 12 neonates. Staffing and maintenance of training in neonatal units are important feasibility considerations, and simple training requirements support the feasibility of the investigational technologies. Key usability characteristics included ease of use, wireless features, and reduced number of attachments connecting the neonate to the monitoring technology, which health care providers considered to increase the efficiency of care. The main factors supporting acceptability included caregiver-highlighted perceptions of neonate comfort and health care respondent technology familiarity. Concerns about the side effects of wireless connections, electromagnetic fields, and mistrust of unfamiliar technologies have emerged as possible acceptability barriers to investigational technologies. Overall, respondents considered the investigational technologies feasible, usable, and acceptable for the care of neonates at this health care facility. Our findings highlight the potential of different multiparameter continuous physiological monitoring technologies for use in different neonatal care settings. Simple and user-friendly technologies may help to bridge gaps in current care where there are many neonates; however, challenges in maintaining training and ensuring feasibility within resource-constrained health care settings warrant further research. RR2-10.1136/bmjopen-2019-035184.


Molecular characterization of a locus required for hyaluronic acid capsule production in group A streptococci.
To characterize the production of hyaluronate capsule by the membrane-associated enzyme hyaluronate synthase (HAS), group A streptococci from a recent outbreak of acute rheumatic fever were mutagenized via Tn916 insertion. Acapsular transconjugants harboring multiple, nontandem copies of the transposon were identified and found to lack HAS activity (less than 1% of wild-type levels). Generalized transduction was then performed to determine which Tn916 insertion was responsible for the HAS- phenotype. These marker exchange experiments resulted in the isolation of two distinct classes of acapsular transductants, designated WF61 and WF62. Both transductants also lacked significant HAS activity, and excision of the transposon from WF62 restored capsular hyaluronate production. Southern analysis of WF61 DNA demonstrated a large deletion of genomic DNA adjacent to the Tn916 insertion. This deletion event is presumably responsible for the observed stability of the acapsular phenotype of WF61. Further analyses of transductant whole-cell DNA indicated that the transposon insertions of WF61 and WF62 were separated by 2.5 kb. These studies define a locus required for hyaluronate capsule production in group A streptococci. Further genetic analysis of this locus has identified a gene required for HAS activity which wasd inactivated by TN916 in WF62 and deleted in WF61.


The M170 reflects a viewpoint-dependent representation for both familiar and unfamiliar faces.
The aim of this study was to determine the extent to which the neural representation of faces in visual cortex is viewpoint dependent or viewpoint invariant. Magnetoencephalography was used to measure evoked responses to faces during an adaptation paradigm. Using familiar and unfamiliar faces, we compared the amplitude of the M170 response to repeated images of the same face with images of different faces. We found a reduction in the M170 amplitude to repeated presentations of the same face image compared with images of different faces when shown from the same viewpoint. To establish if this adaptation to the identity of a face was invariant to changes in viewpoint, we varied the viewing angle of the face within a block. We found a reduction in response was no longer evident when images of the same face were shown from different viewpoints. This viewpoint-dependent pattern of results was the same for both familiar and unfamiliar faces. These results imply that either the face-selective M170 response reflects an early stage of face processing or that the computations underlying face recognition depend on a viewpoint-dependent neuronal representation.


Physical Activity and Cognitive Function in Middle-Aged and Older Adults: An Analysis of 104,909 People From 20 Countries.
To examine the associations of physical activity (PA) frequency (both moderate and vigorous intensity) and PA levels with cognitive function. Data of individuals 50 years or older, from 20 European countries (along with Israel), were collected from 2004 to November 2013 in the biannual Survey of Health, Ageing and Retirement in Europe. A total of 104,909 participants were assessed for cognitive function at least once (mean follow-up length, 29.5±35.7 months). Baseline moderate- and vigorous-intensity PA were reported by participants as more than once a week, once a week, one to three times a month, or hardly ever or never. With regard to PA frequency, participants were categorized as inactive, low active, intermediate active, or high active. The main outcome measure is a cognitive composite score created by summing the z scores of 4-item temporal orientation, 10-word list for delayed recall, and verbal fluency. Adjusted, 2-level mixed-effect regressions found that compared with doing no PA, doing PA more than once a week, once a week, or one to three times a month was positively associated with the composite score (β coefficients varied from 0.52 to 0.75 for moderate-intensity PA and from 0.26 to 0.33 for vigorous-intensity PA). Similarly, compared with the inactive category, high-active, intermediate-active, and low-active categories had positive associations with the composite score (β varied from 0.77 to 1.10). Positive associations were also obtained between PA variables and the raw scores of cognitive tests. Physical activity has dose-response associations with cognitive function, with even low PA frequencies (few times per month) being positively associated with cognitive function during aging.


Allelic variation on chromosome 5 controls beta-cell mass expansion during hyperglycemia in leptin receptor-deficient diabetes mice.
Leptin signaling is a critical component of normal insulin sensitivity. Overt hyperglycemia and type 2 diabetes mellitus can be manifested in states of leptin signaling deficiencies by the additional effects of other genetic factors. We have previously described the contrasting insulin sensitivities and glycemic states of two congenic diabetes (db/db) mouse strains. C57BL/6J db/db mice have mild insulin resistance and achieve euglycemia with mild hyperinsulinemia. FVB db/db mice have severe insulin resistance and are hyperglycemic despite escalating hyperinsulinemia with expanded pancreatic beta-cell mass. Analysis of obese progeny from the two reciprocal backcrosses suggests that genetic modifiers for insulin sensitivity are separable from loci that modulate beta-cell mass. A genome scan of the backcross to FVB suggests that one or more modifier genes are present on chromosome 5. This evidence is supported by the phenotypes of multiple incipient congenic strains wherein the hyperglycemia observed in obese FVB mice is reproduced. With similar degrees of hyperglycemia in obese mice of these strains, the haplotype at chromosome 5 is associated with beta-cell mass and circulating insulin concentrations. Finally, we offer arguments that production of multiple incipient congenic lines is an economical alternative to the production of speed congenic strains.


Memristive Ion Channel-Doped Biomembranes as Synaptic Mimics.
Solid-state neuromorphic systems based on transistors or memristors have yet to achieve the interconnectivity, performance, and energy efficiency of the brain due to excessive noise, undesirable material properties, and nonbiological switching mechanisms. Here we demonstrate that an alamethicin-doped, synthetic biomembrane exhibits memristive behavior, emulates key synaptic functions including paired-pulse facilitation and depression, and enables learning and computing. Unlike state-of-the-art devices, our two-terminal, biomolecular memristor features similar structure (biomembrane), switching mechanism (ion channels), and ionic transport modality as biological synapses while operating at considerably lower power. The reversible and volatile voltage-driven insertion of alamethicin peptides into an insulating lipid bilayer creates conductive pathways that exhibit pinched current-voltage hysteresis at potentials above their insertion threshold. Moreover, the synapse-like dynamic properties of the biomolecular memristor allow for simplified learning circuit implementations. Low-power memristive devices based on stimuli-responsive biomolecules represent a major advance toward implementation of full synaptic functionality in neuromorphic hardware.


Evolving uses of oral reverse transcriptase inhibitors in the HIV-1 epidemic: from treatment to prevention.
The HIV epidemic continues unabated, with no highly effective vaccine and no cure. Each new infection has significant economic, social and human costs and prevention efforts are now as great a priority as global antiretroviral therapy (ART) scale up. Reverse transcriptase inhibitors, the first licensed class of ART, have been at the forefront of treatment and prevention of mother to child transmission over the past two decades. Now, their use in adult prevention is being extensively investigated. We describe two approaches: treatment as prevention (TasP) - the use of combination ART (2NRTI and 1NNRTI) following HIV diagnosis to limit transmission and pre-exposure prophylaxis (PrEP) -the use of single or dual oral agents prior to sexual exposure. Prevention of mother-to-child transmission using NRTI has been highly successful, though does not involve sustained use of NRTI to limit transmission. Despite theoretical and preliminary support for TasP and PrEP, data thus far indicate that adherence, retention in care and late diagnosis are the major barriers to their successful, sustained implementation. Future advances in drug technologies will be needed to overcome the issue of drug adherence, through development of drugs that involve both less frequent dosing as well as reduced toxicity, possibly through specific targeting of infected cells.


Linear recording condition in multirecording telecine system.
Since the multirecording telecine system includes the negative photographic film as a component, it is a nonlinear system. In this paper, the distortions of the three electrical primary color signals generated by the film nonlinearity are analyzed and represented as the function of the film gamma gamma and recording level. The chromaticity errors of the reproduced image are calculated for various values of gamma and recording level k The condition where the color differences of all colors are less than 7.5 is defined as the linear recording condition. The regions of gamma and k to satisfy this linear recording condition are calculated.


Adherens junction remodeling by the Notch pathway in Drosophila melanogaster oogenesis.
Identifying genes involved in the control of adherens junction (AJ) remodeling is essential to understanding epithelial morphogenesis. During follicular epithelium development in Drosophila melanogaster, the main body follicular cells (MBFCs) are displaced toward the oocyte and become columnar. Concomitantly, the stretched cells (StCs) become squamous and flatten around the nurse cells. By monitoring the expression of epithelial cadherin and Armadillo, I have discovered that the rate of AJ disassembly between the StCs is affected in follicles with somatic clones mutant for fringe or Delta and Serrate. This results in abnormal StC flattening and delayed MBFC displacement. Additionally, accumulation of the myosin II heavy chain Zipper is delayed at the AJs that require disassembly. Together, my results demonstrate that the Notch pathway controls AJ remodeling between the StCs and that this role is crucial for the timing of MBFC displacement and StC flattening. This provides new evidence that Notch, besides playing a key role in cell differentiation, also controls cell morphogenesis.


Rifabutin-based 10-day and 14-day triple therapy as a third-line and fourth-line regimen for Helicobacter pylori eradication: A pilot study.
This prospective randomized study was designed to assess the efficacy of 10-day and 14-day rifabutin-based triple therapy as a third- or fourth-line rescue therapy. Patients who failed first- and second-line eradication therapy were enrolled. H. pylori was isolated from gastric biopsy specimens and the rpoB mutation status, a factor of resistance to rifamycins, and minimum inhibitory concentrations (MICs) of rifabutin and amoxicillin were determined. Enrolled patients were randomly assigned to receive 10-day or 14-day eradication therapy with esomeprazole (20 mg, 4 times a day (q.i.d.)), amoxicillin (500 mg, q.i.d.), and rifabutin (300 mg, once a day (q.d.s.)). Poor compliance was defined as intake of <80% of study drugs. Successful H. pylori eradication was confirmed using a [13C] urea breath test or a stool antigen test, 12 weeks after the end of therapy. Twelve patients were assigned to the 10-day group, and 17, to the 14-day group. Intention-to-treat and per-protocol analyses of eradication rates were 83.3% and 81.8% for the 10-day group and 94.1% and 91.7% for the 14-day group, respectively. All patients with rpoB mutation-positive strains (n = 3) showed successful eradication, irrespective of the regimen received. Therapy was stopped due to adverse events in 8.3% and 29.3% of patients in the 10-day and 14-day groups, respectively. Both the 10-day and 14-day therapies were effective as rescue regimens. In particular, the 14-day therapy resulted in successful eradication in over 90% of patients, but the 10-day treatment may be enough to obtain a successful eradication rate, considering the tolerability of therapy.


Tobacco industry tactics with advertisements at the point of sale in Mumbai.
The Cigarettes and Other Tobacco Products (Prohibition of Advertising and Regulations of Trade and Commerce, Production, Supply and Distribution) Act 2003 (COTPA) set out a number of stringent regulations to address tobacco promotion, some of which were revised in 2004. The aim of the study was to monitor the industry tactics at the point of sale with advertising and promotion of tobacco product in Mumbai. The study was carried out by Cancer Patients Aid Association in Mumbai with the help of volunteers. The surveys consisted of two parts, observational information and an interviewer administered questionnaire. Observations like size of board, display of advertisement, backlighting, and use of any promotion were noted. A questionnaire captured information about any incentives from tobacco companies for advertisement and promotion was administered to the vendors who agreed to participate. Study was approved by the Scientific and independent Ethics committee. Total 125 establishments (58 shops, 55 kiosks, 12 other sites) with display boards were surveyed across 5 wards in Mumbai. It was noted that the most common violation was the placements of boards, mainly placed above the shop. The display boards were oversized and few of the advertisements were highlighted with backlights. Out of 125 tobacco vendors surveyed, 107 (85.5%) vendors agreed to answer the questionnaire. We noted that a majority of 67% (84 vendors) stated that they had been approached by tobacco companies to place the signages during the past 5 years post COTPA came into effect. 79 vendors (65 %) admitted to being paid by the tobacco companies. Although the civil society and various non-governmental organizations has casted voice against the industry tactics but ineffective enforcement of the law is a major hurdle. It is likely that cigarette companies will be further able to overcome advertising restrictions by finding loopholes in tobacco legislation unless the decision makers ban it comprehensively as evident in other countries.


Understanding the origin of metal-sulfur vibrations in an oxo-molybdenum dithiolene complex: relevance to sulfite oxidase.
X-ray crystallography and resonance Raman (rR) spectroscopy have been used to further characterize (Tp*)MoO(qdt) (Tp* is hydrotris(3,5-dimethyl-1-pyrazolyl)borate and qdt is 2,3-quinoxalinedithiolene), which represents an important benchmark oxomolybdenum mono-dithiolene model system relevant to various pyranopterin Mo enzyme active sites, including sulfite oxidase. The compound (Tp*)MoO(qdt) crystallizes in the triclinic space group, P1, where a = 9.8424 (7) A, b = 11.2323 (8) A, c = 11.9408 (8) A, alpha = 92.7560 (10) degrees, beta = 98.9530 (10) degrees, and gamma = 104.1680 (10) degrees. The (Tp*)MoO(qdt) molecule exhibits the distorted six-coordinate geometry characteristic of related oxo-Mo(V) systems possessing a single coordinated dithiolene ligand. The first coordination sphere bond lengths and angles in (Tp*)MoO(qdt) are very similar to the corresponding structural parameters for (Tp*)MoO(bdt) (bdt is 1,2-benzenedithiolene). The relatively small inner-sphere structural variations observed between (Tp*)MoO(qdt) and (Tp*)MoO(bdt) strongly suggest that geometric effects are not a major contributor to the significant electronic structural differences reported for these two oxo-Mo(V) dithiolenes. Therefore, the large differences observed in the reduction potential and first ionization energy between the two molecules appear to derive primarily from differences in the effective nuclear charges of their respective sulfur donors. However, a subtle perturbation to Mo-S bonding is implied by the nonplanarity of the dithiolene chelate ring, which is defined by the fold angle. This angular distortion (theta = 29.5 degrees in (Tp*)MoO(qdt); 21.3 degrees in (Tp*)MoO(bdt)) observed between the MoS2 and S-C=C-S planes may contribute to the electronic structure of these oxo-Mo dithiolene systems by controlling the extent of S p-Mo d orbital overlap. In enzymes, the fold angle may be dynamically modulated by the pyranopterin, thereby functioning as a transducer of vibrational energy associated with protein conformational changes directly to the active site via changes in the fold angle. This process could effectively mediate charge redistribution at the active site during the course of atom- and electron-transfer processes. The rR spectrum shows bands at 348 and 407 cm(-1). From frequency analysis of the normal modes of the model, [(NH3)3MoO(qdt)]1+, using the Gaussian03 suite of programs, these bands are assigned as mixed-mode Mo-S vibrations of the five-membered Mo-ditholene core structure. Raman spectroscopy has also provided additional evidence for an in-plane pseudo-sigma dithiolene S-Mo d(xy) covalent bonding interaction in (Tp*)MoO(qdt) and related oxo-Mo-dithiolenes that has implications for electron-transfer regeneration of the active site in sulfite oxidase involving the pyranopterin dithiolene.


Identifying eIF4E-binding protein translationally-controlled transcripts reveals links to mRNAs bound by specific PUF proteins.
eIF4E-binding proteins (4E-BPs) regulate translation of mRNAs in eukaryotes. However the extent to which specific mRNA targets are regulated by 4E-BPs remains unknown. We performed translational profiling by microarray analysis of polysome and monosome associated mRNAs in wild-type and mutant cells to identify mRNAs in yeast regulated by the 4E-BPs Caf20p and Eap1p; the first-global comparison of 4E-BP target mRNAs. We find that yeast 4E-BPs modulate the translation of >1000 genes. Most target mRNAs differ between the 4E-BPs revealing mRNA specificity for translational control by each 4E-BP. This is supported by observations that eap1Δ and caf20Δ cells have different nitrogen source utilization defects, implying different mRNA targets. To account for the mRNA specificity shown by each 4E-BP, we found correlations between our data sets and previously determined targets of yeast mRNA-binding proteins. We used affinity chromatography experiments to uncover specific RNA-stabilized complexes formed between Caf20p and Puf4p/Puf5p and between Eap1p and Puf1p/Puf2p. Thus the combined action of each 4E-BP with specific 3'-UTR-binding proteins mediates mRNA-specific translational control in yeast, showing that this form of translational control is more widely employed than previously thought.


Changes in knowledge of bat rabies and human exposure among United States cavers.
The purpose of the study was to evaluate changes in the knowledge of bat rabies and human exposure among United States cavers during the last decade. A survey was distributed among cavers who attended the National Speleological Society convention in 2000 and those who attended in 2010. In 2000 and 2010, 392 and 108 cavers, respectively, responded to the questionnaire. Eighty-five per cent of respondents in 2000 indicated a bat bite as a risk for rabies compared with all respondents in 2010 (P < 0.0001 controlling for age). The proportion of respondents indicating that they were advised to receive rabies pre-exposure prophylaxis (PreEP) because of caving increased (17% and 29%; P = 0.03 controlling for age). Among these, PreEP was received by 56% and 45%. Although recognition of the risk of rabies exposure from bats is important, the proportion of cavers acting on current recommendations regarding PreEP does not appear to have improved in the past decade.


Electromyography and the evolution of motor control: limitations and insights.
Electromyography (EMG), or the study of muscle activation patterns, has long been used to infer central nervous system (CNS) control of the musculoskeletal system and the evolution of that control. As the activation of the muscles at the level of the periphery is a reflection of the interaction of descending influences and local reflex control, EMG is an important tool in integrated investigations of the evolution of coordination in complex, musculoskeletal systems. Yet, the use of EMG as a tool to understand the evolution of motor control has its limitations. We here review the potential limitations and opportunities of the use of EMG in studying the evolution of motor control in vertebrates and provide original previously unpublished data to illustrate this. The relative timing of activation of a set of muscles can be used to evaluate CNS coordination of the components in a musculoskeletal system. Studies of relative timing reveal task-dependent variability in the recruitment of different populations of muscle fibers (i.e., different fiber types) within a single muscle, and left-right asymmetries in activation that need to be taken into account in comparative studies. The magnitude of muscle recruitment is strongly influenced by the instantaneous demands imposed on the system, and is likely determined by local reflex-control systems. Consequently, using EMG to make meaningful inferences about evolutionary changes in musculoskeletal control requires comparisons across similar functional tasks. Moreover, our data show that inferences about the evolution of motor control are limited in their explanatory power without proper insights into the kinematics and dynamics of a system.


Vaccine-induced polioencephalomyelitis in Scotland.
A six-month-old British female, living in Glasgow was admitted in June 1986 with a four-day history of fever and lower limb weakness following immunisation with oral polio and triple (DTP) vaccines. Examination revealed paralysis of all limbs, facial muscles and right diaphragm, scoliosis, opsoclonus and ocular flutter. Poliovirus types 1, 2 and 3, isolated from her stool specimens were all vaccine-like strains. Her serial serum IgA levels were persistently low and salivary IgA was undetectable. This appears to be the first fully authenticated case of poliovaccine damage in Scotland. It is unclear whether the selective IgA deficiency contributed to her vulnerability. It is essential to investigate elaborately and process viral isolates in every suspected case of acute poliomyelitis so as to determine the dimension and ramifications of poliovaccine damage in the UK population which is known to be rather apprehensive about vaccine dangers.


The zebrafish homologs of SET/I2PP2A oncoprotein: expression patterns and insights into their physiological roles during development.
The oncoprotein SET/I2PP2A (protein phosphatase 2A inhibitor 2) participates in various cellular mechanisms such as transcription, cell cycle regulation and cell migration. SET is also an inhibitor of the serine/threonine phosphatase PP2A, which is involved in the regulation of cell homeostasis. In zebrafish, there are two paralogous set genes that encode Seta (269 amino acids) and Setb (275 amino acids) proteins which share 94% identity. We show here that seta and setb are similarly expressed in the eye, the otic vesicle, the brain and the lateral line system, as indicated by in situ hybridization labeling. Whole-mount immunofluorescence analysis revealed the expression of Seta/b proteins in the eye retina, the olfactory pit and the lateral line neuromasts. Loss-of-function studies using antisense morpholino oligonucleotides targeting both seta and setb genes (MOab) resulted in increased apoptosis, reduced cell proliferation and morphological defects. The morphant phenotypes were partially rescued when MOab was co-injected with human SET mRNA. Knockdown of setb with a transcription-blocking morpholino oligonucleotide (MOb) resulted in phenotypic defects comparable with those induced by setb gRNA (guide RNA)/Cas9 [CRISPR (clustered regularly interspaced short palindromic repeats)-associated 9] injections. In vivo labeling of hair cells showed a significantly decreased number of neuromasts in MOab-, MOb- and gRNA/Cas9-injected embryos. Microarray analysis of MOab morphant transcriptome revealed differential expression in gene networks controlling transcription in the sensory organs, including the eye retina, the ear and the lateral line. Collectively, our results suggest that seta and setb are required during embryogenesis and play roles in the zebrafish sensory system development.


Differences between gridded population data impact measures of geographic access to healthcare in sub-Saharan Africa.
Access to healthcare is imperative to health equity and well-being. Geographic access to healthcare can be modeled using spatial datasets on local context, together with the distribution of existing health facilities and populations. Several population datasets are currently available, but their impact on accessibility analyses is unknown. In this study, we model the geographic accessibility of public health facilities at 100-meter resolution in sub-Saharan Africa and evaluate six of the most popular gridded population datasets for their impact on coverage statistics at different administrative levels. Travel time to nearest health facilities was calculated by overlaying health facility coordinates on top of a friction raster accounting for roads, landcover, and physical barriers. We then intersected six different gridded population datasets with our travel time estimates to determine accessibility coverages within various travel time thresholds (i.e., 30, 60, 90, 120, 150, and 180-min). Here we show that differences in accessibility coverage can exceed 70% at the sub-national level, based on a one-hour travel time threshold. The differences are most notable in large and sparsely populated administrative units and dramatically shape patterns of healthcare accessibility at national and sub-national levels. The results of this study show how valuable and critical a comparative analysis between population datasets is for the derivation of coverage statistics that inform local policies and monitor global targets. Large differences exist between the datasets and the results underscore an essential source of uncertainty in accessibility analyses that should be systematically assessed.


Phase 1 dose-escalation study of CP-690 550 in stable renal allograft recipients: preliminary findings of safety, tolerability, effects on lymphocyte subsets and pharmacokinetics.
CP-690 550 inhibits Janus kinase 3 with nanomolar potency. In this dose-escalation study, we assessed the safety, tolerability, effects on lymphocyte subsets, and pharmacokinetics of CP-690 550 when coadministered with mycophenolate mofetil in stable renal allograft recipients for 28 days. Twenty-eight patients were enrolled. Six patients received CP-690 550 5 mg twice daily (BID), 6 patients received 15 mg BID, 10 patients received 30 mg BID, and 6 patients received placebo. The most frequent adverse events were infections and gastrointestinal (abdominal pain, diarrhea, dyspepsia, and vomiting). CP-690 550 15 mg BID and 30 mg BID were associated with a mean decrease in hemoglobin from baseline of 11% and a mean decrease in absolute natural killer cell counts of 50%. CP-690 550 30 mg BID was also associated with a mean increase in absolute CD19(+) B-lymphocytes of 130%. There were no changes in the number of neutrophils, total lymphocytes, platelets, or CD4(+) or CD8(+) T cells; clinical chemistry; vital signs; or electrocardiograms from the pretreatment baseline. Administration of CP-690 550 without a concomitant calcineurin inhibitor resulted in CP-690 550 exposures consistent with previous studies in nontransplant subjects. Additional dose-ranging studies are warranted to evaluate the safety and efficacy of CP-690 550 in renal transplant recipients over longer treatment duration.


The prevention and reduction of weight loss in an acute tertiary care setting: protocol for a pragmatic stepped wedge randomised cluster trial (the PRoWL project).
Malnutrition, with accompanying weight loss, is an unnecessary risk in hospitalised persons and often remains poorly recognised and managed. The study aims to evaluate a hospital-wide multifaceted intervention co-facilitated by clinical nurses and dietitians addressing the nutritional care of patients, particularly those at risk of malnutrition. Using the best available evidence on reducing and preventing unplanned weight loss, the intervention (introducing universal nutritional screening; the provision of oral nutritional supplements; and providing red trays and additional support for patients in need of feeding) will be introduced by local ward teams in a phased way in a large tertiary acute care hospital. A pragmatic stepped wedge randomised cluster trial with repeated cross section design will be conducted. The unit of randomisation is the ward, with allocation by a random numbers table. Four groups of wards (n = 6 for three groups, n = 7 for one group) will be randomly allocated to each intervention time point over the trial. Two trained local facilitators (a nurse and dietitian for each group) will introduce the intervention. The primary outcome measure is change in patient's body weight, secondary patient outcomes are: length of stay, all-cause mortality, discharge destinations, readmission rates and ED presentations. Patient outcomes will be measured on one ward per group, with 20 patients measured per ward per time period by an unblinded researcher. Including baseline, measurements will be conducted at five time periods. Staff perspectives on the context of care will be measured with the Alberta Context Tool. Unplanned and unwanted weight loss in hospital is common. Despite the evidence and growing concern about hospital nutrition there are very few evaluations of system-wide nutritional implementation programs. This project will test the implementation of a nutritional intervention across one hospital system using a staged approach, which will allow sequential rolling out of facilitation and project support. This project is one of the first evidence implementation projects to use the stepped wedge design in acute care and we will therefore be testing the appropriateness of the stepped wedge design to evaluate such interventions. ACTRN12611000020987.


Attenuating effects of caffeic acid phenethyl ester and betaine on abamectin-induced hepatotoxicity and nephrotoxicity.
Abamectin (ABM) is a widely utilized potent anthelmintic and insecticidal agent. In this study, we investigated the protective effects of caffeic acid phenethyl ester (CAPE) and betaine (BET) against ABM-induced hepatotoxicity and nephrotoxicity in rats. Forty rats were divided into five groups, receiving either oral saline solution (normal control), oral ABM at a dose of 2 mg/kg BW (1/5 LD50), CAPE (10 μmol/kg BW intraperitoneally) followed by ABM, or BET supplementation at a dose of 250 mg/kg BW followed by ABM administration, while group V rats received a combination of i.p. CAPE and oral BET in the same doses before receiving ABM. Biochemical analysis showed that ABM administration significantly (p < 0.05) increased serum levels of aminotransferases, alkaline phosphatase, lactate dehydrogenase, and cholesterol, as well as serum creatinine and urea. Compared to the control group, ABM-intoxicated rats had significantly (p < 0.05) higher tissue concentrations of nitric oxide and malondialdehyde, as well as lower tissue glutathione concentration, total antioxidant capacity, and antioxidant enzymatic activity (glutathione peroxidase, superoxide dismutase, and catalase). Histopathological examination of hepatic and renal tissues of ABM-intoxicated rats showed acute inflammatory and necrotic changes. Pretreatment with CAPE and/or BET reversed the biochemical and histopathological alterations of ABM on the liver and kidneys. Therefore, CAPE and BET (alone or in combination) could be promising protective agents against ABM-induced hepatotoxicity and nephrotoxicity. Future studies should confirm our findings and evaluate the other molecular effects are involved in the combination chemoprotection of CAPE and BET.


Pancreatic Cancer Complicated by Pancreatitis Demonstrated on FDG PET/CT.
A 73-year-old man was referred for F-FDG PET/CT study for staging of biopsy-proven pancreatic adenocarcinoma. The scan demonstrated focal intense FDG uptake in the pancreatic head, localizing the primary tumor. Additional moderate diffuse uptake was seen throughout the pancreas, suggestive of acute pancreatitis. Concurrent diagnostic CT showed diffuse pancreatic hypoenhancement consistent with edema. Serum lipase level was elevated, confirming a diagnosis of acute pancreatitis.


[Recurrent benign multicystic peritoneal mesothelioma: Approach to this rare condition].
Benign multicystic mesothelioma is a rare benign tumour derived from the peritoneal mesothelium. The aim of this paper is to present a case of this rare tumour and review the clinical features, diagnosis and treatment of this disease. The case is presented of a 22-year-old female diagnosed with multicystic mesothelioma after an urgent resection of intra-abdominal tumour in the context of acute abdominal pain. In the subsequent follow-up, the patient had a recurrence of the lesion, and at 2 years was treated by further resection. Benign multicystic mesothelioma is a benign tumour of unknown origin, and with a non-specific clinical manifestation. The most effective treatment is surgical, although there is a high tendency to local recurrence.


Insights into human cognition from intracranial EEG: A review of audition, memory, internal cognition, and causality.
By recording neural activity directly from the human brain, researchers gain unprecedented insight into how neurocognitive processes unfold in real time. We first briefly discuss how intracranial electroencephalography (iEEG) recordings, performed for clinical practice, are used to study human cognition with the spatiotemporal and single-trial precision traditionally limited to non-human animal research. We then delineate how studies using iEEG have informed our understanding of issues fundamental to human cognition: auditory prediction, working and episodic memory, and internal cognition. We also discuss the potential of iEEG to infer causality through the manipulation or 'engineering' of neurocognitive processes via spatiotemporally precise electrical stimulation. We close by highlighting limitations of iEEG, potential of burgeoning techniques to further increase spatiotemporal precision, and implications for future research using intracranial approaches to understand, restore, and enhance human cognition.


Correlations of Self-Reported Androgen Deficiency in Ageing Males (ADAM) with Stress and Sleep among Young Adult Males.
Androgen deficiency in males has traditionally been predominantly limited to older men aged 50+ years. However, little is known of the correlation between hormonal disruption, stress, and sleep in college-aged males. This cross-sectional study investigates lifestyle behavior patterns in young men and a screening for potential androgen deficiency. A survey of 409 male students, as part of a larger USDA-funded GetFruved study, was analyzed for this subproject. Survey instruments used include the Androgen Deficiency in the Aging Male Questionnaire (ADAM) to assess for inadequate ADAM scores, the Perceived Stress Scale to measure stress levels and the Pittsburgh Sleep Quality Index to evaluate sleep quality. In total, 144 male participants (35%) met criteria for potential androgen deficiency defined by the ADAM questionnaire. Correlation was found between having a positive ADAM score and both increased stress levels (p < 0.001) and poor sleep quality (p < 0.001), with stress displaying the strongest effect (p < 0.001 vs p = 0.124). An increased prevalence of having a positive ADAM score versus established norms for this age group was also noted. These findings highlight the need for investigation of endocrine disruptions in young men.


Developmental changes in the sensitivity of the N-methyl-D-aspartate receptor to polyamines.
The binding of [125I]I-MK-801 to N-methyl-D-aspartate (NMDA) receptors on membranes prepared from cultured cerebral cortical neurons and from forebrain of rats of different ages was investigated. Specific binding of [125I]I-MK-801 was enhanced by glutamate, glycine, and polyamines and was inhibited by divalent cations and open-channel blockers of the NMDA receptor, indicating that [125I]I-MK-801 selectively labels a component of the NMDA receptor/ion channel complex. The ability of spermine to enhance the binding of [125I]I-MK-801 was lower in membranes prepared from cultured cerebral cortical neurons or from neonatal rat brain than in membranes prepared from adult rat brain. There was a progressive increase in the potency of spermine and in the magnitude of the stimulatory effect of spermine in rat forebrain between days 3 and 10 of postnatal life. In contrast, the apparent affinity of the NMDA receptor for spermine remained unchanged in cerebral cortical neurons maintained in culture for up to 5 weeks. Mg2+ also enhanced the binding of [125I]I-MK-801 and was more potent in membranes prepared from adult than from 3-day-old rat forebrain. The potency of glutamate for enhancing the binding of [125I]I-MK-801 was not altered in 3-day-old, compared with adult, brain tissue. The increase in the affinity of the polyamine recognition site on the NMDA receptor complex in rat forebrain during the first 2 weeks of postnatal life suggests that the macromolecular properties of the NMDA receptor are altered during development. This may suggest that the subunit composition of the NMDA receptor is under developmental control. Cultured cortical neurons may represent a useful system for investigating factors that regulate developmental changes in the properties of the NMDA receptor.


The association of dyslipoproteinemia with corneal arcus and xanthelasma. The Lipid Research Clinics Program Prevalence Study.
The prevalence rates of xanthelasma and corneal arcus were computed for normolipidemic and dyslipoproteinemic participants in the Lipid Research Clinics population surveys. The prevalence of both xanthelasma and corneal arcus increased with age, was highest in persons with type II phenotype, and usually low in those with type IV phenotype. The relative effects of age and use of gonadal hormones were different in the two lesions. Both xanthelasma and corneal arcus were associated with increased levels of plasma cholesterol and low-density lipoprotein cholesterol (LDL-C), especially in young males. In general, persons with either lesion had increased odds of having type IIa dyslipoproteinemia but decreased odds of having type IV. Xanthelasma and corneal arcus were highly associated with each other, especially in young people. There was no consistent univariate association of xanthelasma and corneal arcus with smoking, alcohol consumption, blood pressure, obesity, sedentary lifestyle, family history of ischemic heart disease, or glucose and uric acid plasma concentrations. Adjusted odds ratios for ischemic heart disease in participants with xanthelasma and corneal arcus were generally increased, except in older female nonusers of gonadal hormones, in whom they were significantly decreased. Neither xanthelasma nor corneal arcus showed a consistent association with manifestations of peripheral arterial disease. Thus the clinical findings of xanthelasma or corneal arcus, especially in young people, seem to identify persons with plasma lipoprotein abnormalities.


Acute myeloid leukemia and clonal chromosome aberrations in relation to past exposure to organic solvents.
The effects of occupational and leisure-time exposures on the risk of acute myeloid leukemia (AML) were investigated with emphasis on clonal chromosome aberrations (CCA) and morphological subtypes. Consecutively diagnosed cases of AML (N=333) and 1 population referent per case were retrospectively included in the study. Information on worktasks, companies, and leisure-time activities was obtained with telephone interviews. Exposure probability and intensity were assessed by occupational hygienists. Associations were evaluated with logistic regression. Exposure to organic solvents was associated with an increased risk of AML [low exposure: OR 1.5 (95% confidence interval (95% CI) 1.0-2.3, moderate-high exposure: OR 2.3 (95% CI 1.0-5.0)]. For exposure to solvents, but not to benzene, the OR was 1.2 (95% CI 0.69-2.0) for "low" and 2.7 (95% CI 1.0-7.3) for "moderate-high" exposure. The observed effects increased with intensity and duration of exposure. The estimated effects were higher for patients >60 years of age at the time of diagnosis. The effect of exposure to organic solvents was not differential with regard to morphology [except possibly erythroleukemia: OR 4.2, 95% CI 1.0-17 or the presence of CCA in general]. No increased risk for AML with complex CCA or with total or partial losses of chromosomes 5 or 7 were observed, but a higher risk was found for AML with trisomy 8 (OR 11, 95% CI 2.7-42) as the sole aberration. Exposure to organic solvents was associated with an increased risk of AML. This association was not due to benzene exposure alone and may be modified by age. Furthermore, specific associations with trisomy 8, and possibly also erythroleukemia, were suggested.


Cellular expression of plasma prekallikrein in human tissues.
Plasma prekallikrein (PPK) is synthesised in hepatocytes and secreted into the blood, where it participates in the surface-dependent activation of blood coagulation, fibrinolysis, kinin generation and inflammation. Recently we demonstrated by quantitative RT-PCR that the human PPK gene is transcribed not only in the liver, but also in various non-hepatic human tissues at significant levels. However, up to now no reliable information is available concerning protein synthesis in the corresponding human tissues. Here we demonstrate by immunohistochemical studies that PPK or plasma kallikrein (PK) is localised in cells of different embryologically derived human tissues. In the human nephron, single cells of the distal tubules stained intensely, while the cytoplasm of cells forming proximal tubules and collecting ducts stained uniformly. PPK/PK was localised in hepatic epithelial cells of the liver, in cells of the pancreatic islet of Langerhans, in the interstitial Leydig cells of the testes, in the follicular and thecal granulosa cells of the ovary, and in the parotid gland, oesophagus, skin, respiratory tract, prostate and breast. We conclude that the cellular localisation of PPK/PK in multiple different progenitor-derived cells indicates specific cellular functions of this enzyme, in addition to its known function in the blood.


Gut permeability, inflammation, and bone density across the menopause transition.
BACKGROUNDInflammation is implicated in many aging-related disorders. In animal models, menopause leads to increased gut permeability and inflammation. Our primary objective was to determine if gut permeability increases during the menopause transition (MT) in women. Our exploratory objectives were to examine whether greater gut permeability is associated with more inflammation and lower bone mineral density (BMD).METHODSWe included 65 women from the Study of Women's Health Across the Nation (SWAN). Key measures were markers of gut permeability (gut barrier dysfunction, fatty acid binding protein 2 [FABP2]) and immune activation secondary to gut microbial translocation (LPS binding protein [LBP], soluble CD14 [sCD14]), inflammation (high-sensitivity CRP), and lumbar spine (LS) or total hip (TH) BMD.RESULTSIn our primary analysis, FABP2, LBP, and sCD14 increased by 22.8% (P = 0.001), 3.7% (P = 0.05), and 8.9% (P = 0.0002), respectively, from pre- to postmenopause. In exploratory, repeated measures, mixed-effects linear regression (adjusted for BMI, age at the premenopausal visit, race/ethnicity, and study site), greater gut permeability was associated with greater inflammation, along with lower LS and TH BMD.CONCLUSIONGut permeability increases during the MT. Greater gut permeability is associated with more inflammation and lower BMD. Future studies should examine the longitudinal associations of gut permeability, inflammation, and BMD.FUNDINGFunding for this research was provided by NIH, Department of Health and Human Services, through the National Institute on Aging, National Institute of Nursing Research, and NIH Office of Research on Women's Health (U01NR004061, U01AG012505, U01AG012535, U01AG012531, U01AG012539, U01AG012546, U01AG012553, U01AG012554, and U01AG012495).


Longitudinal fluctuation of antibodies to extractable nuclear antigens in systemic lupus erythematosus.
To examine the appearance, persistence, and disappearance of anti-extractable nuclear antigen (ENA, Sm, U1-RNP, Ro/SSA, and La/SSB) and anti-native DNA (dsDNA) antibodies during systemic lupus erythematosus (SLE) followup. One hundred and thirty patients who fulfilled American College of Rheumatology classification criteria for SLE with at least 5 yearly anti-ENA and dsDNA tests between 1987-2002 were retrospectively selected. Four longitudinal antibody data patterns were considered for each antibody: always absent, always present, absent at diagnosis with positive seroconversion, and present at diagnosis with negative seroconversion. Antibodies to Ro/SSA were present in 47%, U1-RNP in 36%, DNA in 32%, Sm in 23%, and La/SSB in 7% of patients. Among patients ever positive for a given autoantibody, the frequency of the "always present" pattern was 52% for anti-Ro/SSA, 38% for U1-RNP, 17% for Sm, 11% for La/SSB, and 9% for DNA antibodies; the frequency of positive seroconversion was 56% for anti-La/SSB, 33% for DNA, 26% for Sm, 21% for U1-RNP, and 15% for Ro/SSA. Time to positive seroconversion varied from 1 to 8 years after diagnosis. Among patients with a positive test at diagnosis the frequency of those remaining positive between the 2nd and 4th year of followup decreased to 39-78%, depending upon autoantibody specificity; between the 5th and 10th years this rate was 20-75%. Antibody data pattern frequency differed significantly among autoantibody specificities except between anti-U1-RNP and Ro/SSA (p = 0.15) and between anti-DNA and Sm antibodies (p = 0.29). The high frequency of longitudinal fluctuation in anti-ENA antibodies suggests that a periodic reappraisal may be appropriate in seronegative patients with a suspect diagnosis of SLE. The clinical significance of such fluctuation deserves future study.


Endovascular stent graft repair of abdominal and thoracic aortic aneurysms: a ten-year experience with 817 patients.
On November 23, 1992, the first endovascular stent graft (ESG) repair of an aortic aneurysm was performed in North America. Following the treatment of this patient, we have continued to evaluate ESG over the past 10 years in the treatment of 817 patients. Abdominal (AAA) or thoracic (TAA) aortic aneurysms are a significant health concern traditionally treated by open surgical repair. ESG therapy may offer protection from aneurysm rupture with a reduction in procedure morbidity and mortality. Over a 10-year period, 817 patients were treated with ESGs for AAA (723) or TAA (94). Patients received 1 of 12 different stent graft devices. Technical and clinical success of ESGs was reviewed, and the incidence of procedure-related complications was analyzed. The mean age was 74.3 years (range, 25-95 years); 678 patients (83%) were men; 86% had 2 or more comorbid medical illnesses, 67% of which included coronary artery disease. Technical success, on an intent-to-treat basis was achieved in 93.8% of patients. Primary clinical success, which included freedom from aneurysm-related death, type I or III endoleak, graft infection or thrombosis, rupture, or conversion to open repair was 65 +/- 6% at 8 years. Of great importance, freedom from aneurysm rupture after ESG insertion was 98 +/- 1% at 9 years. There was a 2.3% incidence of perioperative mortality. One hundred seventy five patients died of causes not related to their aneurysm during a mean follow-up of 15.4 months. Stent graft therapy for aortic aneurysms is a valuable alternative to open aortic repair, especially in older sicker patients with large aneurysms. Continued device improvements coupled with an enhanced understanding of the important role of aortic pathology in determining therapeutic success will eventually permit ESGs to be a more durable treatment of aortic aneurysms.


Sexual dimorphism in the inflammatory response to traumatic brain injury.
The activation of resident microglial cells, alongside the infiltration of peripheral macrophages, are key neuroinflammatory responses to traumatic brain injury (TBI) that are directly associated with neuronal death. Sexual disparities in response to TBI have been previously reported; however it is unclear whether a sex difference exists in neuroinflammatory progression after TBI. We exposed male and female mice to moderate-to-severe controlled cortical impact injury and studied glial cell activation in the acute and chronic stages of TBI using immunofluorescence and in situ hybridization analysis. We found that the sex response was completely divergent up to 7 days postinjury. TBI caused a rapid and pronounced cortical microglia/macrophage activation in male mice with a prominent activated phenotype that produced both pro- (IL-1β and TNFα) and anti-inflammatory (Arg1 and TGFβ) cytokines with a single-phase, sustained peak from 1 to 7 days. In contrast, TBI caused a less robust microglia/macrophage phenotype in females with biphasic pro-inflammatory response peaks at 4 h and 7 days, and a delayed anti-inflammatory mRNA peak at 30 days. We further report that female mice were protected against acute cell loss after TBI, with male mice demonstrating enhanced astrogliosis, neuronal death, and increased lesion volume through 7 days post-TBI. Collectively, these findings indicate that TBI leads to a more aggressive neuroinflammatory profile in male compared with female mice during the acute and subacute phases postinjury. Understanding how sex affects the course of neuroinflammation following brain injury is a vital step toward developing personalized and effective treatments for TBI.


Direct combinatorial interaction between a herpes simplex virus regulatory protein and a cellular octamer-binding factor mediates specific induction of virus immediate-early gene expression.
We provide evidence for a novel mechanism of transcriptional regulation in which the immediate-early (IE) transactivating protein of herpes simplex virus, Vmw65, is assembled into a specific DNA-binding complex together with a cellular octamer-binding factor (TRF). The assembly of Vmw65/TRF complex requires not only the core TRF recognition site, but also flanking sequences which are dispensable for TRF binding alone. We show from functional analyses that TRF binding by a motif is required but not sufficient to confer induction on a heterologous promoter, and it is the ability of the motif to allow TRF/Vmw65 complex assembly which correlates with functional activity. Thus, for the induction of HSV IE expression, Vmw65 forms a complex with TRF by recognition of the specific subset of appropriately flanked TRF binding sites present in each of the IE genes. This mechanism may provide a paradigm for the selective utilization of the same transcription factor in differential gene expression.


The influence of insertion torque on the survival of immediately placed and restored single-tooth implants.
The purpose of this retrospective study was to evaluate the medium- to long-term clinical outcome of single-tooth implants placed into fresh extraction sockets using a low-insertion-torque protocol and immediately restored with acrylic resin provisional crowns. Sixty-one patients consecutively referred for the replacement of one or more failing teeth were planned for treatment by means of extraction and immediate implant placement and provisionalization using a low-insertion-torque protocol of ≤ 25 Ncm. Low rotational stability was not a contraindication to treatment unless there was a lack of axial stability. Implant survival and marginal bone levels were documented. A statistical analysis was performed to discover any correlations between insertion torque, age, gender, implant size and tooth position and marginal bone loss. Sixty-eight implants were placed. Three implants failed to osseointegrate; one was replaced and immediately restored and has been successful for more than 5 years. The overall survival rate for all implants was 95.5%, which have been in function for a period of 1.25 to 9.5 years. The mean marginal bone loss of the 54 implants followed for at least 24 months was 0.23 ± 0.60 mm mesially and 0.20 ± 0.72 mm distally. Overall, 78% of implants showed no marginal bone loss, 9% experienced 0.1 to 0.5 mm of bone loss, and 13% demonstrated > 0.5 mm of bone loss. There were no significant correlations between the various parameters analyzed and marginal bone loss. A torque of only 25 Ncm would seem more than sufficient to yield a favorable clinical outcome, and there may be a misconception as to what actually represents adequate primary stability. Immediate provisionalization of single-tooth implants placed with a relatively low insertion torque can yield favorable survival rates and optimal maintenance of marginal bone levels compared to the generally accepted norm.


Tissue Tracking: Applications for Brain MRI Classification.
Bayesian classification methods have been extensively used in a variety of image processing applications, including medical image analysis. The basic procedure is to combine data-driven knowledge in the likelihood terms with clinical knowledge in the prior terms to classify an image into a pre-determined number of classes. In many applications, it is difficult to construct meaningful priors and, hence, homogeneous priors are assumed. In this paper, we show how expectation-maximization weights and neighboring posterior probabilities may be combined to make intuitive use of the Bayesian priors. Drawing upon insights from computer vision tracking algorithms, we cast the problem in a tissue tracking framework. We show results of our algorithm on the classification of gray and white matter along with surrounding cerebral spinal fluid in brain MRI scans. We show results of our algorithm on 20 brain MRI datasets along with validation against expert manual segmentations.


Who abandons embryos after IVF?
This investigation describes features of in vitro fertilisation (IVF) patients who never returned to claim their embryos following cryopreservation. Frozen embryo data were reviewed to establish communication patterns between patient and clinic; embryos were considered abandoned when 1) an IVF patient with frozen embryo/s stored at our facility failed to make contact with our clinic for > 2 yrs and 2) the patient could not be located after a multi-modal outreach effort was undertaken. For these patients, telephone numbers had been disconnected and no forwarding address was available. Patient, spouse and emergency family contact/s all escaped detection efforts despite an exhaustive public database search including death records and Internet directory portals. From 3244 IVF cycles completed from 2000 to 2008, > or = 1 embryo was frozen in 1159 cases (35.7%). Those without correspondence for > 2 yrs accounted for 292 (25.2%) patients with frozen embryos; 281 were contacted by methods including registered (signature involving abandoned embryos did not differ substantially from other patients. The goal of having a baby was achieved by 10/11 patients either by spontaneous conception, adoption or IVF. One patient moved away with conception status unconfirmed. The overall rate of embryo abandonment was 11/1159 (< 1%) in this IVF population. Pre-IVF counselling minimises, but does not totally eliminate, the problem of abandoned embryos. As the number of abandoned embryos from IVF accumulates, their fate urgently requires clarification. We propose that clinicians develop a policy consistent with relevant Irish Constitutional provisions to address this medical dilemma.


Haplotypes spanning centromeric regions reveal persistence of large blocks of archaic DNA.
Despite critical roles in chromosome segregation and disease, the repetitive structure and vast size of centromeres and their surrounding heterochromatic regions impede studies of genomic variation. Here we report the identification of large-scale haplotypes (cenhaps) in humans that span the centromere-proximal regions of all metacentric chromosomes, including the arrays of highly repeated α-satellites on which centromeres form. Cenhaps reveal deep diversity, including entire introgressed Neanderthal centromeres and equally ancient lineages among Africans. These centromere-spanning haplotypes contain variants, including large differences in α-satellite DNA content, which may influence the fidelity and bias of chromosome transmission. The discovery of cenhaps creates new opportunities to investigate their contribution to phenotypic variation, especially in meiosis and mitosis, as well as to more incisively model the unexpectedly rich evolution of these challenging genomic regions.


Open Science principles for accelerating trait-based science across the Tree of Life.
Synthesizing trait observations and knowledge across the Tree of Life remains a grand challenge for biodiversity science. Species traits are widely used in ecological and evolutionary science, and new data and methods have proliferated rapidly. Yet accessing and integrating disparate data sources remains a considerable challenge, slowing progress toward a global synthesis to integrate trait data across organisms. Trait science needs a vision for achieving global integration across all organisms. Here, we outline how the adoption of key Open Science principles-open data, open source and open methods-is transforming trait science, increasing transparency, democratizing access and accelerating global synthesis. To enhance widespread adoption of these principles, we introduce the Open Traits Network (OTN), a global, decentralized community welcoming all researchers and institutions pursuing the collaborative goal of standardizing and integrating trait data across organisms. We demonstrate how adherence to Open Science principles is key to the OTN community and outline five activities that can accelerate the synthesis of trait data across the Tree of Life, thereby facilitating rapid advances to address scientific inquiries and environmental issues. Lessons learned along the path to a global synthesis of trait data will provide a framework for addressing similarly complex data science and informatics challenges.


The impact of metabolic supply lines - and the patterns between them - on the development of distant metastases in 64 women with breast cancer.
Cancer cells upregulate their metabolism to underlie the increased malignant activity. This requires an increased amount of 'metabolic building materials', for example glucose, amino acids etc., which have the blood circulation as their principal supply lines. Targeting these metabolic supply lines, and thus the availability of metabolic building materials in the blood, therefore carries treatment potential. A central observation is that the malignant alterations comprise great complexity and that compensatory mechanisms exist. Therefore, targeted supply lines should presumably constitute specific patterns to achieve therapeutic effect. The aim of the present study was to investigate if such patterns could be seen to correlate with the development of distant metastases. The study was conducted using a case-cohort design. In total, 64 women diagnosed with breast cancer between January 2011 and December 2015 were included. Among these, 32 had developed distant metastases and 32 had not. From a blood sample drawn at the time of diagnosis, the levels of glucose (HbA1c), glutamine, arginine and cystathionine were measured. Cox regression was applied to investigate the impact of the supply lines of these 'building materials' and specifically the patterns between them on the development of distant metastases. The results demonstrate a significant impact of the investigated metabolic supply lines, centrally in relation to interaction between them and in relation to the impact of the increased cumulated utilization of multiple supply lines simultaneously. In conclusion, the results indicated that the metabolic supply lines may impact clinical outcome, and, in this regard, the results placed a substantial emphasis on the effect of the patterns between these supply lines.


Specific binding sites for an antifungal plant defensin from Dahlia (Dahlia merckii) on fungal cells are required for antifungal activity.
Dm-AMP1, an antifungal plant defensin from seeds of dahlia (Dahlia merckii), was radioactively labeled with t-butoxycarbonyl-[35S]-L-methionine N-hydroxy-succinimi-dylester. This procedure yielded a 35S-labeled peptide with unaltered antifungal activity. [35S]Dm-AMP1 was used to assess binding on living cells of the filamentous fungus Neurospora crassa and the unicellular fungus Saccharomyces cerevisiae. Binding of [35S]Dm-AMP1 to fungal cells was saturable and could be competed for by preincubation with excess, unlabeled Dm-AMP1 as well as with Ah-AMP1 and Ct-AMP1, two plant defensins that are highly homologous to Dm-AMP1. In contrast, binding could not be competed for by more distantly related plant defensins or structurally unrelated antimicrobial peptides. Binding of [35S]Dm-AMP1 to either N. crassa or S. cerevisiae cells was apparently irreversible. In addition, whole cells and microsomal membrane fractions from two independently obtained S. cerevisiae mutants selected for resistance to Dm-AMP1 exhibited severely reduced binding affinity for [35S]Dm-AMP1, compared with wild-type yeast. This finding suggests that binding of Dm-AMP1 to S. cerevisiae plasma membranes is required for antifungal activity of this protein.


First Report of Apricot pseudo-chlorotic leaf spot virus Infecting Plum (Prunus domestica) in the Czech Republic.
Apricot pseudo-chlorotic leaf spot virus (APCLSV) is a novel, still poorly known Trichovirus in the family Betaflexiviridae. It is most closely related to Apple chlorotic leaf spot virus (ACLSV) (2,4) and infects stone fruit trees of the Prunus genus. Its presence has so far been detected in apricot, plum, Japanese plum, and peach trees in Italy, Spain, France, Hungary, Turkey, Jordan, and Australia (1,2,4). During the summers of 2008 and 2010, leaf samples of old Czech local plum cultivars were obtained from the Holovousy collection and assessed for the presence of viruses belonging to the Capillovirus, Trichovirus, and Foveavirus genera using the polyvalent degenerate oligonucleotides (PDO) nested reverse transcription (RT)-PCR test (3). Following amplification from total RNAs extracts, the amplicons were cloned and several clones were sequenced for each plant sample. In plum (Prunus domestica) cv. Babce, a mixture of amplicons was observed and BlastN and BlastX analyses of the obtained sequences revealed the presence of ACLSV and APCLSV. The 310-bp APCLSV amplicon (GenBank Accession No. JN790294) showed highest identity (82.9% in nucleotide sequence and 97.1% in amino acid sequence) with the Sus2 isolate of APCLSV (4) and clustered with APCLSV isolates in a phylogenetic analysis. APCLSV infection was further confirmed with an APCLSV-specific RT-PCR assay (4), which yielded a product of the expected 205-bp size (GenBank Accession No. JN653070) with closest homology again to the Sus2 APCLSV isolate (83.4 and 94.3% nucleotide and amino acid identity, respectively). To our knowledge, this finding represents the first detection of APCLSV in domestic plums in the Czech Republic, extending our vision of APCLSV diversity and its geographic distribution. For unknown reasons, APCLSV has almost always been reported in mixed infection with ACLSV (1,2,4) and the situation in cv. Babce does not deviate from this trend. This has greatly hindered the analysis of the pathogenicity of APCLSV, a situation further complicated in the current case because the Babce cultivar was also infected by Plum pox virus. References: (1) M. Barone et al. Acta Hortic. 781:53, 2008. (2) T. Candresse et al. Virus and Virus-Like Diseases of Pome and Stone Fruit Trees. A. Hadidi et al., eds. The American Phytopathological Society, St. Paul, MN, 2011. (3) X. Foissac et al. Phytopathology 95:617, 2005. (4) D. Liberti et al. Phytopathology 95:420, 2005.


Apical closure device for transapical valve procedures.
Transapical transcatheter valve procedures are performed through a left minithoracotomy and require apical sutures to seal the apical access site. The use of large-calibre devices compromises any attempt to fully perform the procedure with a thoracoscopic approach or percutaneously. We report our preliminary experience in animals with a new sutureless self-expandable apical occluder, engineered to perform transapical access site closure in a minimally invasive setting with large-size introducer sheaths. The apical occluder with extendable waist was implanted in six young pigs during an acute animal study. Under general anaesthesia, animals (mean weight: 62 ± 8 kg) received full heparinization (heparin: 100 UI/kg; activated clotting time above 250 s). Through a median sternotomy, a 21-Fr Certitude™ introducer sheath (outer diameter: 25 Fr) was placed over the wire into the cardiac apex. The delivery catheter carrying the constrained apical plug was inserted into the sheath and deployed under fluoroscopic control, whereas the Certitude™ was retrieved. After protamine infusion, we observed and recorded the 1-h bleeding with standard haemodynamic parameters. Animals were sacrificed, and hearts analysed. Six apical closure devices were successfully introduced and deployed in six pig hearts through large-size apical sheaths at first attempt. In all animals, the plugs guaranteed immediate apical sealing and traces of blood were collected in the pericardium during the 1-h observational period (mean of 16 ± 3.4 ml of blood loss per animal). Haemodynamic parameters remained stable during the entire study period and no plug dislodgement was detected with normal systemic blood pressure (mean arterial mean blood pressure: 65 ± 7 mmHg). Post-mortem analysis confirmed the full deployment and good fixation of all plugs, without macroscopic damages to the surrounding myocardium. This sutureless self-expandable apical occluder is a simple device capable of sealing large-size apical access sites (20-35 Fr) in an acute animal study. This approach is a step further towards less invasive transapical valve procedures in the clinical setting, and further animal tests will be performed to confirm the long-term efficacy and safety of this device.


Effects of an enzymatically stable C-terminal hexapseudopeptide fragment peptide of xenin-25, ψ-xenin-6, on pancreatic islet function and metabolism.
Xenin-25 undergoes rapid enzyme metabolism following secretion. Early studies demonstrated bioactivity of a C-terminal hexapeptide fragment of xenin-25, namely xenin-6, which were enhanced through introduction of a reduced N-terminal peptide bond, to yield Ψ-xenin-6. The present study was undertaken to define the biological actions and potential antidiabetic properties of Ψ-xenin-6. In vitro enzymatic stability, insulin and glucagon secretory activity, as well as effects on beta-cell survival were determined. Studies in mice were used to assess the impact of Ψ-xenin-6 on glucose homeostasis and satiety. Ψ-xenin-6 was resistant to murine plasma degradation. In BRIN-BD11 cells and isolated murine islets, Ψ-xenin-6 significantly stimulated insulin secretion, and prominently enhanced the insulinotropic actions of GIP. Xenin-6 and Ψ-xenin-6 had no impact on glucagon secretion, although xenin-6 partially reversed the glucagonotropic action of GIP. Further in vitro investigations revealed that, similar to GLP-1, Ψ-xenin-6 significantly augmented proliferation of human and rodent clonal beta-cells, whilst also fully protecting against cytokine-induced beta-cell cytotoxicity, with greater potency than xenin-25 and xenin-6. When administered to mice in combination with glucose, Ψ-xenin-6 significantly reduced glucose levels and enhanced glucose-induced insulin release, with a duration of biological action beyond 8 h. Ψ-xenin-6 also significantly enhanced the glucose-lowering action of GIP in vivo. In overnight fasted mice, Ψ-xenin-6 exhibited satiety actions at both 25 and 250 nmol/kg. These data demonstrates that Ψ-xenin-6 is a metabolically stable C-terminal fragment analogue of xenin-25, with a metabolic action profile that merits further study as a potential antidiabetic compound.