The autoantibody-mediated encephalitides: from clinical observations to molecular pathogenesis.
The autoimmune encephalitis (AE) syndromes have been characterised by the detection of autoantibodies in serum and/or cerebrospinal fluid which target the extracellular domains of specific neuroglial antigens. The clinical syndromes have phenotypes which are often highly characteristic of their associated antigen-specific autoantibody. For example, the constellation of psychiatric features and the multi-faceted movement disorder observed in patients with NMDAR antibodies are highly distinctive, as are the faciobrachial dystonic seizures observed in close association with LGI1 antibodies. These typically tight correlations may be conferred by the presence of autoantibodies which can directly access and modulate their antigens in vivo. AE remains an under-recognised clinical syndrome but one where early and accurate detection is critical as prompt initiation of immunotherapy is closely associated with improved outcomes. In this review of a rapidly emerging field, we outline molecular observations with translational value. We focus on contemporary methodologies of autoantibody detection, the evolution and distinctive nature of the clinical phenotypes, generalisable therapeutic paradigms, and finally discuss the likely mechanisms of autoimmunity in these patients which may inform future precision therapies.


Clinical laboratory automation: concepts and designs.
Compared with other industries, automation in the healthcare arena has been slow to evolve. Changes in reimbursement for services provided are forcing hospitals and other healthcare providers to look for more cost effective mechanisms to provide all forms of health related services. Clinical laboratory services are essential to the support of hospital operations and most clinic operations. The current paradigm for clinical laboratory operations is based upon a mix of both batch and random access testing and is highly dependent upon clinical laboratory personnel. Changes in the paradigm for clinical laboratory operations may result in a significant cost savings when fully implemented while maintaining a high level of quality and service for the patients. Several different laboratory organization structures are discussed. The Nebraska automation project is described including the development of a modular conveyor system, the use of automated guided vehicles, software, and network architectures. Future directions for the development of clinical laboratory operations including the creation of a "docking device" to link automation systems to instruments is proposed.


Knowledge and involvement of husbands in maternal and newborn health in rural Bangladesh.
Access to skilled health services during pregnancy, childbirth and postnatal period for obstetric care is one of the strongest determinants of maternal and newborn health (MNH) outcomes. In many countries, husbands are key decision-makers in households, effectively determining women's access to health services. We examined husbands' knowledge and involvement regarding MNH issues in rural Bangladesh, and how their involvement is related to women receiving MNH services from trained providers. We conducted a cross-sectional survey in two rural sub-districts of Bangladesh in 2014 adopting a stratified cluster sampling technique. Women with a recent birth history and their husbands were interviewed separately with a structured questionnaire. A total of 317 wife-husband dyads were interviewed. The associations between husbands accompanying their wives as explanatory variables and utilization of skilled services as outcome variables were assessed using multiple logistic regression analyses. In terms of MNH knowledge, two-thirds of husbands were aware that women have special rights related to pregnancy and childbirth and one-quarter could mention three or more pregnancy-, birth- and postpartum-related danger signs. With regard to MNH practice, approximately three-quarters of husbands discussed birth preparedness and complication readiness with their wives. Only 12% and 21% were involved in identifying a potential blood donor and arranging transportation, respectively. Among women who attended antenatal care (ANC), 47% were accompanied by their husbands. Around half of the husbands were present at the birthplace during birth. Of the 22% women who received postpartum care (PNC), 67% were accompanied by their husbands. Husbands accompanying their wives was positively associated with women receiving ANC from a medically trained provider (AOR 4.5, p < .01), birth at a health facility (AOR 1.5, p < .05), receiving PNC from a medically trained provider (AOR 48.8, p < .01) and seeking care from medically trained providers for obstetric complications (AOR 3.0, p < 0.5). Husbands accompanying women when receiving health services is positively correlated with women's use of skilled MNH services. Special initiatives should be taken for encouraging husbands to accompany their wives while availing MNH services. These initiatives should aim to increase men's awareness regarding MNH issues, but should not be limited to this.


Conformational effects, molecular orbitals, and reaction activities of bis(phthalocyaninato) lanthanum double-deckers: density functional theory calculations.
The conformational effects on the frontier molecular orbital energy and stability for reduced, neutral, and oxidized bis(phthalocyaninato) lanthanum double-deckers have been revealed on the basis of density functional theory calculations. Calculation results indicate that the frontier orbital coupling degree changes along with the molecular conformation of the double-decker compound, first decreasing along with the increase of rotation angle β from 0 to 20° and then increasing along with the increase of rotation angle β from 20 to 45°. In addition, the stability for the three forms of double-decker changes in the same order, but first increasing and then decreasing along with the change of the rotation angle β in the range of 0 to 45° with a rotation energy barrier of (31.3 ± 3.1) kJ mol(-1) at 20°. This reveals that the rotation of the two phthalocyanine rings for the reduced, neutral, and oxidized bis(phthalocyaninato) lanthanum double-deckers are able to occur at room temperature. Nevertheless, the superior coordination reaction activity of the neutral bis(phthalocyaninato) lanthanum double-decker complex over their reduced form in forming sandwich-type tris(phthalocyaninato) lanthanum triple-decker compounds has also been clearly clarified on the basis of comparative calculations on the Fukui function of [La(Pc)(2)] and [La(Pc)(2)](-) using the DFT method. Fukui function analysis reveals the reaction center of the 18-electron-π-conjugated core in the bis(phthalocyaninato) lanthanum double-decker molecule against both electrophilic and radical attack. Nevertheless, the larger global chemical softness (S) for the neutral [La(Pc)(2)] than the reduced form [La(Pc)(2)](-) indicates the higher reaction activity of the former form over the latter one. This explains well the experimental findings that only the neutral instead of the reduced form of bis(tetrapyrrole) rare earth double-decker complexes, containing at least one phthalocyanine ligand, could be employed as starting materials towards the preparation of tris(tetrapyrrole) rare earth triple-decker complexes by a solution process.


Bismuth@US-tubes as a Potential Contrast Agent for X-ray Imaging Applications.
The encapsulation of bismuth as BiOCl/Bi2O3 within ultra-short (ca. 50 nm) single-walled carbon nanocapsules (US-tubes) has been achieved. The Bi@US-tubes have been characterized by high-resolution transmission electron microscopy (HR-TEM), energy-dispersive X-ray spectroscopy (EDS), thermogravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy. Bi@US-tubes have been used for intracellular labeling of pig bone marrow-derived mesenchymal stem cells (MSCs) to show high X-ray contrast in computed tomography (CT) cellular imaging for the first time. The relatively high contrast is achieved with low bismuth loading (2.66% by weight) within the US-tubes and without compromising cell viability. X-ray CT imaging of Bi@US-tubes-labeled MSCs showed a nearly two-fold increase in contrast enhancement when compared to unlabeled MSCs in a 100 kV CT clinical scanner. The CT signal enhancement from the Bi@US-tubes is 500 times greater than polymer-coated Bi2S3 nanoparticles and several-fold that of any clinical iodinated contrast agent (CA) at the same concentration. Our findings suggest that the Bi@US-tubes can be used as a potential new class of X-ray CT agent for stem cell labeling and possibly in vivo tracking.


Riboswitch structure in the ligand-free state.
Molecular investigations of riboswitches bound to small-molecule effectors have produced a wealth of information on how these molecules achieve high affinity and specificity for a target ligand. X-ray crystal structures have been determined for the ligand-free state for representatives of the preQ₁-I, S-adenosylmethionine I, lysine, and glycine aptamer classes. These structures in conjunction with complimentary techniques, such as in-line probing, NMR spectroscopy, Förster resonance energy transfer, small-angle scattering, and computational simulations, have demonstrated that riboswitches adopt multiple conformations in the absence of ligand. Despite a number of investigations that support ligand-dependent folding, mounting evidence suggests that free-state riboswitches interact with their effectors in the sub-populations of largely prefolded states as embodied by the principle of conformational selection, which has been documented extensively for protein-mediated ligand interactions. Fundamental riboswitch investigations of the bound and free states have advanced our understanding of RNA folding, ligand recognition, and how these factors culminate in communication between an aptamer and its expression platform. An understanding of these topics is essential to comprehend riboswitch gene regulation at the molecular level, which has already provided a basis to understand the mechanism of action of natural antimicrobials.


AUJESZKY'S DISEASE IN HUNTED WILD BOAR (SUS SCROFA) IN THE IBERIAN PENINSULA.
Aujeszky's disease (AD, pseudorabies) eradication programs in domestic pigs are implemented in several European countries where AD virus (ADV) circulates in local wild boar (Sus scrofa), making studies on ADV infection dynamics in wild boar increasingly relevant. The objective of our study was to characterize ADV dynamics in wild boar at a site in central Portugal and compare this site to three enzootic sites in central Spain. A total of 235 wild boar were sampled during the hunting season 2014-15. We collected serum, tissues (oropharyngeal tonsils and trigeminal and sacral ganglia), and swabs (oral, nasal, and genital) and analyzed these samples to detect ADV antibodies (enzyme-linked immunosorbent assay) and DNA (PCR). An overall seroprevalence of 42.6% was found (range 12.7-57.7%), being highest in adults (54.1%; 72/133). Overall, 2.8% (3/108) oral, 6.4% (7/109) nasal, and 12.8% (12/94) genital swabs were PCR positive. We found 20.4% (20/98) of the wild boar had at least one positive swab and were considered shedders. We found ADV in tissues of five animals; of 111 tonsils, three (2.7%) were PCR positive. Trigeminal (2/48; 4%) and sacral (2/53; 4%) ganglia collected in central Portugal, pertaining to three animals, were positive for ADV DNA. Logistic regression models showed that seroprevalence was influenced by site and age, whereas ADV shedding was influenced by site. Our study describes patterns of ADV infection in wild boar in Portugal and shows that wild boar also pose a risk, albeit lower than that in central Spain, for the eradication of AD from extensively managed domestic pigs in Portugal.


Beyond patency: Functional assessment of adequacy using internal mammary artery grafting to the left anterior descending artery.
Arterial graft physiology influences the long-term outcome of coronary artery bypass grafting (CABG). We studied factors that can affect the overall resistance to flow using internal mammary artery grafting to the left anterior descending artery. This was a prospective, nonrandomized observational study of 100 consecutive patients who underwent elective on-pump isolated or combined valve surgery and CABG. Coronary stenoses were assessed using conventional and quantitative coronary angiography assessment. The flow and pulsatility index (PI) of the grafts were assessed by transit-time flowmetry during cardioplegic arrest and at the end of the operation. Fractional polynomials were used to explore linearity, followed by multivariable regression analysis. Univariate analysis demonstrated higher flows at the end of the operation in patients who had higher flows with the cross-clamp on (P < .001), in males (P = .004), in patients with a low PI at the end of the operation (P = .04), and in patients with a larger size of the recipient artery (P = .005). Multivariable regression analysis showed that the graft flow at the end of the operation was significantly associated with the mean flow with the cross-clamp on (P < .001), sex (P = .003), and PI at the end of the operation (P = .003). Concomitant valve surgery did not influence flows. Male patients had 18 mL/min higher flow. The graft flow at the end of the operation can be determined by the flow with the cross-clamp on, the PI with the cross-clamp off and coronary artery. We reported differences in the graft flows between sexes, and for first the time, we introduced the concepts of "adequate flow" and "resistance-to-forward-flow" for patent coronary grafts.


Imaging zinc: old and new tools.
As recently as 20 years ago, all zinc in biological systems was believed to be tightly bound to proteins, and the idea of imaging zinc was considered heretical. Beginning with Maske's research with dithizonate staining of the hippocampus in the 1950s, however, zinc-sensitive dyes have indicated that, in mammalian cells, free zinc can exist in at least three separate pools. These pools include vesicular zinc sequestered in presynaptic vesicles and secretory granules, zinc released from these vesicles into the extracellular space after physiological stimulation, and transient increases in zinc in cells in the regions where extracellular release of zinc has occurred. This Perspective covers the zinc-imaging tools, from dithizonate to the newest FRET-based sensors, that have galvanized biomedical science.


The epidemiology of MRI detected shoulder injuries in athletes participating in the Rio de Janeiro 2016 Summer Olympics.
To use Magnetic Resonance Imaging (MRI) to characterize the severity, location, prevalence, and demographics of shoulder injuries in athletes at the Rio de Janeiro 2016 Summer Olympic Games. This was a retrospective analysis of all routine shoulder MRIs obtained from the Olympic Village Polyclinic during the Rio 2016 Summer Olympics. Imaging was performed on 1.5 T and 3 T MRI, and interpretation was centrally performed by a board-certified musculoskeletal radiologist. Images were assessed for tendon, muscle, bone, bursal, joint capsule, labral, and chondral abnormality. A total of 11,274 athletes participated in the Games, of which 55 (5%) were referred for a routine shoulder MRI. Fifty-three (96%) had at least two abnormal findings. Seven (13%) had evidence of an acute or chronic anterior shoulder dislocation. Forty-nine (89%) had a rotator cuff partial tear and / or tendinosis. Subacromial / subdeltoid bursitis was present in 29 (40%). Thirty (55%) had a tear of the superior labrum anterior posterior (SLAP). Our study demonstrated a high prevalence of both acute and chronic shoulder injuries in the Olympic athletes receiving shoulder MRI. The high rates of bursal, rotator cuff, and labral pathology found in these patients implies that some degree of glenohumeral instability and impingement is occurring, likely due to fatigue and overuse of the dynamic stabilizers. Future studies are needed to better evaluate sport-specific trends of injury.


Assessing the risk of intussusception and rotavirus vaccine safety in Canada.
Intussusception has been identified as a rare adverse event following rotavirus immunization. We sought to determine the incidence of intussusception among infants in Canada both before and after introduction of rotavirus immunization programs. We used Canadian Institute for Health Information (CIHI) Discharge Abstract Database (DAD) to identify infants under 1 y of age who were admitted to a Canadian hospital, which the exception of Quebec, which does not submit data to CIHI, with a diagnosis of intussusception (ICD-10 code K56.1, and ICD-9 code 560) between January 1st, 2003 and December 31, 2013. We compared rates of intussusception hospitalization before and after rotavirus vaccine program introduction. Rates were adjusted for calendar year, age (in months), sex and region using Poisson regression models. Denominator data for infants under 1 year, stratified by age in months, were obtained from Statistics Canada. Annual intussusception hospitalization rates ranged from 20-30 per 100,000 infants over the study period, with no evidence of a trend over time. Intussusception hospitalization rates were highest in infants 4 to <8 months and lowest in those under 2 months or between 10 and <12 months. Males had higher rates than females both overall and within each age group. The rate of intussusception hospitalization after rotavirus vaccine program introduction was 22.4 (95% CI: 18.3, 27.4) compared to 23.4 (95% CI: 21.5, 25.4) per 100,000 before program introduction. We have described baseline intussusception hospitalization rates for infants in Canada and have found no evidence of a change in rate after implementation of routine rotavirus immunization programs.


State Entropy and Differentiation Phenomenon.
In the formalism of quantum theory, a state of a system is represented by a density operator. Mathematically, a density operator can be decomposed into a weighted sum of (projection) operators representing an ensemble of pure states (a state distribution), but such decomposition is not unique. Various pure states distributions are mathematically described by the same density operator. These distributions are categorized into classical ones obtained from the Schatten decomposition and other, non-classical, ones. In this paper, we define the quantity called the state entropy. It can be considered as a generalization of the von Neumann entropy evaluating the diversity of states constituting a distribution. Further, we apply the state entropy to the analysis of non-classical states created at the intermediate stages in the process of quantum measurement. To do this, we employ the model of differentiation, where a system experiences step by step state transitions under the influence of environmental factors. This approach can be used for modeling various natural and mental phenomena: cell's differentiation, evolution of biological populations, and decision making.


Foveated near-eye display using computational holography.
Holographic display is the only technology that can offer true 3D with all the required depth cues. Holographic head-worn displays (HWD) can provide continuous depth planes with the correct stereoscopic disparity for a comfortable 3D experience. Existing HWD approaches have small field-of-view (FOV) and small exit pupil size, which are limited by the spatial light modulator (SLM). Conventional holographic HWDs are limited to about 20° × 11° FOV using a 4 K SLM panel and have fixed FOV. We present a new optical architecture that can overcome those limitations and substantially extend the FOV supported by the SLM. Our architecture, which does not contain any moving parts, automatically follows the gaze of the viewer's pupil. Moreover, it mimics human vision by providing varying resolution across the FOV resulting in better utilization of the available space-bandwidth product of the SLM. We propose a system that can provide 28° × 28° instantaneous FOV within an extended FOV (the field of view that is covered by steering the instantaneous FOV in space) of 60° × 40° using a 4 K SLM, effectively providing a total enhancement of > 3 × in instantaneous FOV area, > 10 × in extended FOV area and the space-bandwidth product. We demonstrated 20° × 20° instantaneous FOV and 40° × 20° extended FOV in the experiments.


MutS HOMOLOG1 is a nucleoid protein that alters mitochondrial and plastid properties and plant response to high light.
Mitochondrial-plastid interdependence within the plant cell is presumed to be essential, but measurable demonstration of this intimate interaction is difficult. At the level of cellular metabolism, several biosynthetic pathways involve both mitochondrial- and plastid-localized steps. However, at an environmental response level, it is not clear how the two organelles intersect in programmed cellular responses. Here, we provide evidence, using genetic perturbation of the MutS Homolog1 (MSH1) nuclear gene in five plant species, that MSH1 functions within the mitochondrion and plastid to influence organellar genome behavior and plant growth patterns. The mitochondrial form of the protein participates in DNA recombination surveillance, with disruption of the gene resulting in enhanced mitochondrial genome recombination at numerous repeated sequences. The plastid-localized form of the protein interacts with the plastid genome and influences genome stability and plastid development, with its disruption leading to variegation of the plant. These developmental changes include altered patterns of nuclear gene expression. Consistency of plastid and mitochondrial response across both monocot and dicot species indicate that the dual-functioning nature of MSH1 is well conserved. Variegated tissues show changes in redox status together with enhanced plant survival and reproduction under photooxidative light conditions, evidence that the plastid changes triggered in this study comprise an adaptive response to naturally occurring light stress.


Common alleles at 6q25.1 and 1p11.2 are associated with breast cancer risk for BRCA1 and BRCA2 mutation carriers.
Two single nucleotide polymorphisms (SNPs) at 6q25.1, near the ESR1 gene, have been implicated in the susceptibility to breast cancer for Asian (rs2046210) and European women (rs9397435). A genome-wide association study in Europeans identified two further breast cancer susceptibility variants: rs11249433 at 1p11.2 and rs999737 in RAD51L1 at 14q24.1. Although previously identified breast cancer susceptibility variants have been shown to be associated with breast cancer risk for BRCA1 and BRCA2 mutation carriers, the involvement of these SNPs to breast cancer susceptibility in mutation carriers is currently unknown. To address this, we genotyped these SNPs in BRCA1 and BRCA2 mutation carriers from 42 studies from the Consortium of Investigators of Modifiers of BRCA1/2. In the analysis of 14 123 BRCA1 and 8053 BRCA2 mutation carriers of European ancestry, the 6q25.1 SNPs (r(2) = 0.14) were independently associated with the risk of breast cancer for BRCA1 mutation carriers [hazard ratio (HR) = 1.17, 95% confidence interval (CI): 1.11-1.23, P-trend = 4.5 × 10(-9) for rs2046210; HR = 1.28, 95% CI: 1.18-1.40, P-trend = 1.3 × 10(-8) for rs9397435], but only rs9397435 was associated with the risk for BRCA2 carriers (HR = 1.14, 95% CI: 1.01-1.28, P-trend = 0.031). SNP rs11249433 (1p11.2) was associated with the risk of breast cancer for BRCA2 mutation carriers (HR = 1.09, 95% CI: 1.02-1.17, P-trend = 0.015), but was not associated with breast cancer risk for BRCA1 mutation carriers (HR = 0.97, 95% CI: 0.92-1.02, P-trend = 0.20). SNP rs999737 (RAD51L1) was not associated with breast cancer risk for either BRCA1 or BRCA2 mutation carriers (P-trend = 0.27 and 0.30, respectively). The identification of SNPs at 6q25.1 associated with breast cancer risk for BRCA1 mutation carriers will lead to a better understanding of the biology of tumour development in these women.


Maslinic acid as an effective anticancer agent.
Maslinic acid (2α,3β-dihydroxyolean-12-en-28-oic acid) is a naturally occurring pentacyclic triterpenic compound. Maslinic acid is gradually gaining attention as an excellent pharmacologically active product because of its premium biological properties. In this review we will focus on chemopreventive properties of Maslinic acid against different cancers. Seemingly, available data is limited and we have yet to unravel how Maslinic acid therapeutically targeted oncogenic cell signal transduction cascades in different cancers. Moreover, there are visible knowledge gaps about the ability of Maslinic acid to modulate oncogenic and tumor suppressor microRNAs in various cancers.


Potential mechanisms of endogenous retroviral-mediated genomic instability in human cancer.
Malignancy results from a complex combination of genetic and epigenetic changes, the full effects of which are still largely unknown. Here we summarize current knowledge of the origin, retrotranspositional activity, epigenetic state, and transcription of human endogenous retroviruses (HERVs), and then discuss the potential effects of their deregulation in cancer. Evidence suggests that cancer-associated epigenetic changes most likely underlie potential HERV-mediated effects on genome and transcriptome instability and may play a role in malignancy. Despite our currently limited understanding of the importance of HERVs or other transposable elements in cancer development, we believe that the emerging era of high-throughput sequencing of cancer genomes, epigenomes, and transcriptomes will provide unprecedented opportunities to investigate these roles in the future.


Biomimetic control based on a model of chemotaxis in Escherichia coli.
In the field of molecular biology, extending now to the more comprehensive area of systems biology, the development of computer models for synthetic cell simulation has accelerated extensively and has begun to be used for various purposes, such as biochemical analysis. These models, describing the highly efficient environmental searching mechanisms and adaptability of living organisms, can be used as machine-control algorithms in the field of systems engineering. To realize this biomimetic intelligent control, we require a stripped-down model that expresses a series of information-processing tasks from stimulation input to movement. Here we selected the bacterium Escherichia coli as a target organism because it has a relatively simple molecular and organizational structure, which can be characterized using biochemical and genetic analyses. We particularly focused on a motility response known as chemotaxis and developed a computer model that includes not only intracellular information processing but also motor control. After confirming the effectiveness and validity of the proposed model by a series of computer simulations, we applied it to a mobile robot control problem. This is probably the first study showing that a bacterial model can be used as an autonomous control algorithm. Our results suggest that many excellent models proposed thus far for biochemical purposes can be applied to problems in other fields.


Expression profiling of tomato pre-abscission pedicels provides insights into abscission zone properties including competence to respond to abscission signals.
Detachment of plant organs occurs in abscission zones (AZs). During plant growth, the AZ forms, but does not develop further until the cells perceive abscission-promoting signals and initiate detachment. Upon signal perception, abscission initiates immediately; if there is no signal, abscission is not induced and the organ remains attached to the plant. However, little attention has been paid to the genes that maintain competence to respond to the abscission signal in the pre-abscission AZ. Recently, we found that the tomato (Solanum lycopersicum) transcription factors BLIND (Bl), GOBLET (GOB), Lateral suppressor (Ls) and a tomato WUSCHEL homologue (LeWUS) are expressed specifically in pre-abscission tissue, the anthesis pedicel AZs. To advance our understanding of abscission, here we profiled genome-wide gene expression in tomato flower pedicels at the pre-abscission stage. We examined the transcriptomes of three tomato flower pedicel regions, the AZ and flanking proximal- (Prox) and distal- (Dis) regions, and identified 89 genes that were preferentially expressed in the AZ compared to both Prox and Dis. These genes included several transcription factors that regulate apical or axillary shoot meristem activity. Also, genes associated with auxin activity were regulated in a Prox-Dis region-specific manner, suggesting that a gradient of auxin exists in the pedicel. A MADS-box gene affecting floral transition was preferentially expressed in the Prox region and other MADS-box genes for floral organ identification were preferentially expressed in Dis, implying that the morphologically similar Prox and Dis regions have distinct identities. We also analyzed the expression of known regulators; in anthesis pedicels, Bl, GOB, Ls and LeWUS were expressed in the vascular cells of the AZ region. However, after an abscission signal, Bl was up-regulated, but GOB, Ls and LeWUS were down-regulated, suggesting that Bl may be a positive regulator of abscission, but the others may be negative regulators. This study reveals region-specific gene expression in tomato flower pedicels at anthesis and identifies factors that may determine the physiological properties of the pre-abscission pedicel. The region-specific transcriptional regulators and genes for auxin activity identified here may prevent flower abscission in the absence of signal or establish competence to respond to the abscission signal.


Two Beclin 1-binding proteins, Atg14L and Rubicon, reciprocally regulate autophagy at different stages.
Beclin 1, a protein essential for autophagy, binds to hVps34/Class III phosphatidylinositol-3-kinase and UVRAG. Here, we have identified two Beclin 1 associated proteins, Atg14L and Rubicon. Atg14L and UVRAG bind to Beclin 1 in a mutually exclusive manner, whereas Rubicon binds only to a subpopulation of UVRAG complexes; thus, three different Beclin 1 complexes exist. GFP-Atg14L localized to the isolation membrane and autophagosome, as well as to the ER and unknown puncta. Knockout of Atg14L in mouse ES cells caused a defect in autophagosome formation. GFP-Rubicon was localized at the endosome/lysosome. Knockdown of Rubicon caused enhancement of autophagy, especially at the maturation step, as well as enhancement of endocytic trafficking. These data suggest that the Beclin 1-hVps34 complex functions in two different steps of autophagy by altering the subunit composition.


Overuse of Chest CT in Patients With Stage I and II Breast Cancer: An Opportunity to Increase Guidelines Compliance at an NCCN Member Institution.
Background: The NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines) recommend that patients with clinical stage I/II breast cancer undergo advanced imaging for staging only when symptomatic. Regardless, many asymptomatic patients undergo chest CT. The goal of this study was to assess the use and results of chest CT in these patients at an NCCN Member Institution. Methods: Patients with breast cancer diagnosed between 1998 and 2012 were identified in a prospectively maintained database. All patients with clinical stage I/II disease who did not receive neoadjuvant chemotherapy were included. Data collected included demographics, tumor size, node status, chest CT within 6 months of diagnosis, imaging findings, need for additional workup, and identification of metastatic disease. Appropriate statistical tests were used for analysis. Results: From 1998 to 2012, 3,321 patients were diagnosed with early-stage breast cancer. Of these, 2,062 (62.1%) had clinical stage I breast cancer at diagnosis and 1,259 (37.9%) had stage II; 227 patients (11%) with stage I and 456 (36.2%) with stage II breast cancer received staging chest CT. Of patients undergoing CT, 184 (26.9%) were found to have pulmonary nodules, which measured ≤5 mm for 128 patients (69.6%), 5 to 10 mm for 46 patients (25.0%), 11 to 20 mm for 6 patients (3.2%), and ≥20 mm for 4 patients (2.2%). Patients undergoing chest CT for staging subsequently underwent a mean of 2.34 (range, 0-16) additional CTs in follow-up. Of all patients undergoing chest CT for staging, only 9 (1.3%) were ultimately diagnosed with pulmonary metastases at an average of 25 months (range, 0-97) after initial staging chest CT. Conclusions: A significant percentage of patients with stage I/II breast cancer underwent unnecessary chest CT as part of their initial workup. Nearly one-third of these patients were found to have pulmonary nodules, but only 1.3% were ever diagnosed with pulmonary metastases. Adherence to NCCN Guidelines will reduce the excessive use of CT chest imaging.


Estimating parametric distributions of storage time and temperature of ready-to-eat foods for U.S. households.
Home refrigeration temperatures and product storage times are important factors for controlling the growth of Listeria monocytogenes in refrigerated ready-to-eat foods. In 2005, RTI International, in collaboration with Tennessee State University and Kansas State University, conducted a national survey of U.S. adults to characterize consumers' home storage and refrigeration practices for 10 different categories of refrigerated ready-to-eat foods. No distributions of storage time or refrigeration temperature were presented in any of the resulting publications. This study used classical parametric survival modeling to derive parametric distributions from the RTI International storage practices data set. Depending on the food category, variability in product storage times was best modeled using either exponential or Weibull distributions. The shape and scale of the distributions varied greatly depending on the food category. Moreover, the results indicated that consumers tend to keep a product that is packaged by a manufacturer for a longer period of time than a product that is packaged at retail. Refrigeration temperatures were comparable to those previously reported, with the variability in temperatures best fit using a Laplace distribution, as an alternative to the empirical distribution. In contrast to previous research, limited support was found for a correlation between storage time and temperature. The distributions provided in this study can be used to better model consumer behavior in future risk assessments.


Measuring atopy in a multi-centre epidemiological study.
Skin prick tests are used as a measure of atopy in epidemiological studies, but results may be influenced by the fieldworker performing the test. In a multi-centre epidemiological study the method of reporting the results should consider the need for comparability of findings from different centres. Data on over 1000 subjects from three English centres of the European Community Respiratory Health Survey were analysed to determine whether allergen wheal should be adjusted for histamine wheal, and what cutoff diameter gave the most comparable results. No consistent relation between allergen wheal diameter and histamine wheal diameter was found for any fieldworker or allergen. A cutoff of > 0 mm for a positive result gave a more consistent relation with the corresponding specific IgE value between fieldworkers than either a cutoff of > or = 3 mm or the use of the mean wheal diameter. While this result is not immediately generalisable to studies using different skin prick test reagents, the method of analysis to determine the appropriate criterion of reporting can be used in other epidemiological studies.


Role of viral and host factors in interferon based therapy of hepatitis C virus infection.
The current standard of care (SOC) for hepatitis C virus (HCV) infection is the combination of pegylated interferon (PEG-IFN), Ribavirin and protease inhibitor for HCV genotype 1. Nevertheless, this treatment is successful only in 70-80% of the patients. In addition, the treatment is not economical and is of immense physical burden for the subject. It has been established now, that virus-host interactions play a significant role in determining treatment outcomes. Therefore identifying biological markers that may predict the treatment response and hence treatment outcome would be useful. Both IFN and Ribavirin mainly act by modulating the immune system of the patient. Therefore, the treatment response is influenced by genetic variations of the human as well as the HCV genome. The goal of this review article is to summarize the impact of recent scientific advances in this area regarding the understanding of human and HCV genetic variations and their effect on treatment outcomes. Google scholar and PubMed have been used for literature research. Among the host factors, the most prominent associations are polymorphisms within the region of the interleukin 28B (IL28B) gene, but variations in other cytokine genes have also been linked with the treatment outcome. Among the viral factors, HCV genotypes are noteworthy. Moreover, for sustained virological responses (SVR), variations in core, p7, non-structural 2 (NS2), NS3 and NS5A genes are also important. However, all considered single nucleotide polymorphisms (SNPs) of IL28B and viral genotypes are the most important predictors for interferon based therapy of HCV infection.


The effect of hypocapnia on skeletal muscle microcirculatory blood flow, oxygenation and pH.
Hypocapnia is known to decrease blood flow and tissue oxygen tension in the brain and the splanchnic organs, but there are few and contradictory results in skeletal muscle. The aim of the present study was to investigate the effect of hypocapnia on microcirculatory blood flow, local skeletal muscle PO2 (PtO2) and pH (pHt). Blood flow was measured using laser-Doppler flowmetry (LDF) and a multichannel antimony microelectrode served for PtO2 and pHt measurements. These parameters were studied in six anesthetized and mechanically ventilated rabbits during normocapnia (arterial PCO2 4.4 kPa), hypocapnia (PCO2 2.3 kPa) and finally a second period of normocapnia (PCO2 4.5 kPa). During normocapnia the relative LDF flux was 100 +/- 53% (mean +/- SD), decreased to 80 +/- 48% (p < 0.01) during hypocapnia and returned to 103 +/- 51% during the second period of normocapnia (N.S.). The PtO2 was 3.2 +/- 1.2 kPa, 2.9 +/- 0.8 kPa (p < 0.05) and 3.2 +/- 1.0 kPa (N.S.), respectively. pHt remained unchanged, although there was a significant arterial alkalosis (pH increased 0.10 units, p < 0.01). These results show that hypocapnia induces a 20% decrease in LDF flow and a 9% reduction in PtO2 with an unchanged pHt level. In this skeletal muscle model, the decrease in microcirculatory blood flow due to vasoconstriction leads to a reduction in both tissue oxygenation and in the removal of acid metabolites, which counteract a developing tissue alkalosis.


Oxidative damage in the kidney induced by 900-MHz-emitted mobile phone: protection by melatonin.
The mobile phones emitting 900-MHz electromagnetic radiation (EMR) may be mainly absorbed by kidneys because they are often carried in belts. Melatonin, the chief secretory product of the pineal gland, was recently found to be a potent free radical scavenger and antioxidant. The aim of this study was to examine 900-MHz mobile phone-induced oxidative stress that promotes production of reactive oxygen species (ROS) on renal tubular damage and the role of melatonin on kidney tissue against possible oxidative damage in rats. The animals were randomly grouped as follows: 1) sham-operated control group and 2) study groups: i) 900-MHz EMR exposed (30 min/day for 10 days) group and ii) 900-MHz EMR exposed+melatonin (100 microg kg(-1) s.c. before the daily EMR exposure) treated group. Malondialdehyde (MDA), an index of lipid peroxidation), and urine N-acetyl-beta-d-glucosaminidase (NAG), a marker of renal tubular damage were used as markers of oxidative stress-induced renal impairment. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities were studied to evaluate the changes of antioxidant status. In the EMR-exposed group, while tissue MDA and urine NAG levels increased, SOD, CAT, and GSH-Px activities were reduced. Melatonin treatment reversed these effects as well. In this study, the increase in MDA levels of renal tissue and in urine NAG and also the decrease in renal SOD, CAT, GSH-Px activities demonstrated the role of oxidative mechanism induced by 900-MHz mobile phone exposure, and melatonin, via its free radical scavenging and antioxidant properties, ameliorated oxidative tissue injury in rat kidney. These results show that melatonin may exhibit a protective effect on mobile phone-induced renal impairment in rats.


Long-Term Effect of Biological Soil Disinfestation on Verticillium Wilt.
Biological soil disinfestation (BSD), involving incorporation of grass combined with plastic mulching, eliminates many soilborne pests and diseases through the creation of anaerobic conditions. BSD was compared at two locations with a nontreated control, Italian ryegrass amendment alone, and plastic mulch alone. After the soil treatments, plots were cropped with Acer platanoides and Catalpa bignonioides and grown for 4 years. Relative to the control, soil inoculum levels of Verticillium dahliae were reduced by 85% after BSD and did not increase for 4 years. Populations of Pratylenchus fallax, known for their interaction with V. dahliae, in the soil and in roots were reduced by 95 to 99%. The incidence of infection by V. dahliae was reduced by 80 to 90%. Verticillium wilt severity was significantly reduced in A. platanoides in all 4 years at one location and in the first 2 years at the other location, and significantly fewer plants died at one location. Shoot length and trunk width were larger after BSD compared with the control at one location. Market value of the crop in BSD plots was up to € 140,000 ha-1 higher for A. platanoides and up to € 190,000 ha-1 higher for C. bignonioides than in the untreated control. BSD is an effective, economically profitable, and environmentally friendly control method for tree nurseries.


State-dependent biasing method for importance sampling in the weighted stochastic simulation algorithm.
The weighted stochastic simulation algorithm (wSSA) was developed by Kuwahara and Mura [J. Chem. Phys. 129, 165101 (2008)] to efficiently estimate the probabilities of rare events in discrete stochastic systems. The wSSA uses importance sampling to enhance the statistical accuracy in the estimation of the probability of the rare event. The original algorithm biases the reaction selection step with a fixed importance sampling parameter. In this paper, we introduce a novel method where the biasing parameter is state-dependent. The new method features improved accuracy, efficiency, and robustness.


HdhQ111 Mice Exhibit Tissue Specific Metabolite Profiles that Include Striatal Lipid Accumulation.
The HTT CAG expansion mutation causes Huntington's Disease and is associated with a wide range of cellular consequences, including altered metabolism. The mutant allele is expressed widely, in all tissues, but the striatum and cortex are especially vulnerable to its effects. To more fully understand this tissue-specificity, early in the disease process, we asked whether the metabolic impact of the mutant CAG expanded allele in heterozygous B6.HdhQ111/+ mice would be common across tissues, or whether tissues would have tissue-specific responses and whether such changes may be affected by diet. Specifically, we cross-sectionally examined steady state metabolite concentrations from a range of tissues (plasma, brown adipose tissue, cerebellum, striatum, liver, white adipose tissue), using an established liquid chromatography-mass spectrometry pipeline, from cohorts of 8 month old mutant and wild-type littermate mice that were fed one of two different high-fat diets. The differential response to diet highlighted a proportion of metabolites in all tissues, ranging from 3% (7/219) in the striatum to 12% (25/212) in white adipose tissue. By contrast, the mutant CAG-expanded allele primarily affected brain metabolites, with 14% (30/219) of metabolites significantly altered, compared to wild-type, in striatum and 11% (25/224) in the cerebellum. In general, diet and the CAG-expanded allele both elicited metabolite changes that were predominantly tissue-specific and non-overlapping, with evidence for mutation-by-diet interaction in peripheral tissues most affected by diet. Machine-learning approaches highlighted the accumulation of diverse lipid species as the most genotype-predictive metabolite changes in the striatum. Validation experiments in cell culture demonstrated that lipid accumulation was also a defining feature of mutant HdhQ111 striatal progenitor cells. Thus, metabolite-level responses to the CAG expansion mutation in vivo were tissue specific and most evident in brain, where the striatum featured signature accumulation of a set of lipids including sphingomyelin, phosphatidylcholine, cholesterol ester and triglyceride species. Importantly, in the presence of the CAG mutation, metabolite changes were unmasked in peripheral tissues by an interaction with dietary fat, implying that the design of studies to discover metabolic changes in HD mutation carriers should include metabolic perturbations.


Field and Laboratory Evaluation of the Microsporidian Parasite Heterosporis sutherlandae: Prevalence, Severity, and Transmission.
Heterosporis sutherlandae is an invasive microsporidian parasite in the Great Lakes region of North America that infects the skeletal muscle of numerous fish species, rendering the fillet unfit for human consumption. Although H. sutherlandae has been identified as a pathogen of concern by state management agencies, there is little information to inform regulation and intervention. We sampled fishes over 1 year from three lakes in northern Minnesota with known infected populations to determine the importance of host demographic and environmental variables for influencing H. sutherlandae infection prevalence. Heterosporis sutherlandae was present during all sampling periods, ranging in prevalence from 1% to 11%. The prevalence of H. sutherlandae among Yellow Perch Perca flavescens varied significantly according to season, with winter having the lowest prevalence (1%) and summer having the highest prevalence (11%). For other fish species, the prevalence of H. sutherlandae also varied significantly with season: the lowest prevalence occurred during spring (1%) and the highest prevalence occurred in fall (9%). Rates of pathogen transmission were estimated by exposing Fathead Minnows Pimephales promelas in the laboratory. Transmission rates were 23% when naïve fish were fed infected tissues and only 2% when naïve fish were held in cohabitation with tissue-fed fish. Exposure method and exposure duration (d) increased the probability that a fish was infected with H. sutherlandae. These findings suggest that H. sutherlandae transmission is greater when a susceptible host consumes infected tissue than when the fish is exposed to spores present in the water column. The current rates of infection in wild fishes are in stark contrast to the prevalence documented in 2004 (28%), suggesting a reduction in H. sutherlandae prevalence within at least one Yellow Perch population in the Laurentian Great Lakes region since the early 2000s.


Multiple roles for Med12 in vertebrate endoderm development.
In zebrafish, the endoderm originates at the blastula stage from the most marginal blastomeres. Through a series of complex morphogenetic movements and differentiation events, the endodermal germ layer gives rise to the epithelial lining of the digestive tract as well as its associated organs such as the liver, pancreas, and swim bladder. How endodermal cells differentiate into distinct cell types such as hepatocytes or endocrine and exocrine pancreatic cells remains a major question. In a forward genetic screen for genes regulating endodermal organ development, we identified mutations at the shiri locus that cause defects in the development of a number of endodermal organs including the liver and pancreas. Detailed phenotypic analyses indicate that these defects are partially due to a reduction in endodermal expression of the hairy/enhancer of split-related gene, her5, at mid to late gastrulation stages. Using the Tg(0.7her5:EGFP)(ne2067) line, we show that her5 is expressed in the endodermal precursors that populate the pharyngeal region as well as the organ-forming region. We also find that knocking down her5 recapitulates some of the endodermal phenotypes of shiri mutants, further revealing the role of her5 in endoderm development. Positional cloning reveals that shiri encodes Med12, a regulatory subunit of the transcriptional Mediator complex recently associated with two human syndromes. Additional studies indicate that Med12 modulates the ability of Casanova/Sox32 to induce sox17 expression. Thus, detailed phenotypic analyses of embryos defective in a component of the Mediator complex have revealed new insights into discrete aspects of vertebrate endoderm development, and provide possible explanations for the craniofacial and digestive system defects observed in humans with mutations in MED12.


Metagenomic and functional analysis of hindgut microbiota of a wood-feeding higher termite.
From the standpoints of both basic research and biotechnology, there is considerable interest in reaching a clearer understanding of the diversity of biological mechanisms employed during lignocellulose degradation. Globally, termites are an extremely successful group of wood-degrading organisms and are therefore important both for their roles in carbon turnover in the environment and as potential sources of biochemical catalysts for efforts aimed at converting wood into biofuels. Only recently have data supported any direct role for the symbiotic bacteria in the gut of the termite in cellulose and xylan hydrolysis. Here we use a metagenomic analysis of the bacterial community resident in the hindgut paunch of a wood-feeding 'higher' Nasutitermes species (which do not contain cellulose-fermenting protozoa) to show the presence of a large, diverse set of bacterial genes for cellulose and xylan hydrolysis. Many of these genes were expressed in vivo or had cellulase activity in vitro, and further analyses implicate spirochete and fibrobacter species in gut lignocellulose degradation. New insights into other important symbiotic functions including H2 metabolism, CO2-reductive acetogenesis and N2 fixation are also provided by this first system-wide gene analysis of a microbial community specialized towards plant lignocellulose degradation. Our results underscore how complex even a 1-microl environment can be.


The Relation of Obesity-Related Hormonal and Cytokine Levels With Multiple Myeloma and Non-Hodgkin Lymphoma.
This article presents the first detailed overview of the mechanisms that may underlie the relation of obesity with B-cell non-Hodgkin lymphomas (NHLs) and multiple myeloma (MM). Epidemiologic studies, including meta-analyses of prospective cohorts, have reported that the risks of NHL and MM are significantly increased in obese, relative to normal weight, women and men. Accumulating experimental and clinical evidence suggests that inflammatory cytokines, hyperinsulinemia, and sex hormones could play a role in the association of obesity with B-cell NHL and MM carcinogenesis. There is, however, a paucity of data published from appropriate large prospective cohort studies, and studies concurrently measuring these correlated factors, to formally determine the likely biologic factors driving the relationship of obesity with NHL and MM. Additional strengths and weaknesses of the current literature, as well as study design issues that need to be considered in conducting these studies, such as the exclusion of type 2 diabetics or postmenopausal women using hormone therapy, are discussed.


Long-distance autumn migration across the Sahara by painted lady butterflies: exploiting resource pulses in the tropical savannah.
The painted lady, Vanessa cardui, is a migratory butterfly that performs an annual multi-generational migration between Europe and North Africa. Its seasonal appearance south of the Sahara in autumn is well known and has led to the suggestion that it results from extremely long migratory flights by European butterflies to seasonally exploit the Sahel and the tropical savannah. However, this possibility has remained unproven. Here, we analyse the isotopic composition of butterflies from seven European and seven African countries to provide new support for this hypothesis. Each individual was assigned a geographical natal origin, based on its wing stable hydrogen isotope (δ2Hw) value and a predicted δ2Hw basemap for Europe and northern Africa. Natal assignments of autumn migrants collected south of the Sahara confirmed long-distance movements (of 4000 km or more) starting in Europe. Samples from Maghreb revealed a mixed origin of migrants, with most individuals with a European origin, but others having originated in the Sahel. Therefore, autumn movements are not only directed to northwestern Africa, but also include southward and northward flights across the Sahara. Through this remarkable behaviour, the productive but highly seasonal region south of the Sahara is incorporated into the migratory circuit of V. cardui.


Replication and meta-analysis of candidate loci identified variation at RAB3GAP1 associated with keratoconus.
Keratoconus is a common complex corneal ectasia that can lead to severe visual impairment. Although a genetic component is well recognized, the genetic risk factors for keratoconus are yet to be fully elucidated. A recent genome-wide association study (GWAS) by Li et al. identified 15 potentially associated single nucleotide polymorphisms (SNPs). Here, we aimed to replicate these associations, and conduct a meta-analysis of the current and previous studies. We genotyped the 15 reported associated SNPs in 524 Australian Caucasian cases with keratoconus and 2761 controls. Association analysis was conducted in PLINK. A meta-analysis of this study with the adjusted P values of the previously published GWAS was conducted using the method of Fisher to combine P values. Our Australian cohort showed association (P < 0.003) at SNPs near RAB3GAP1, KCND3, IMMPL2, and in a gene desert on chromosome 13q33.3, providing evidence of replication of the published results. The meta-analysis showed SNP rs4954218 near RAB3GAP1 gene was associated significantly with keratoconus, with P = 9.26 × 10(-9) passing the genome-wide significance level. Although the mechanism of disease association is yet to be determined, SNP rs4954218 is associated consistently with keratoconus and likely tags a functional variant that contributes to disease susceptibility.


A structurally conserved motif in γ-herpesvirus uracil-DNA glycosylases elicits duplex nucleotide-flipping.
Efficient γ-herpesvirus lytic phase replication requires a virally encoded UNG-type uracil-DNA glycosylase as a structural element of the viral replisome. Uniquely, γ-herpesvirus UNGs carry a seven or eight residue insertion of variable sequence in the otherwise highly conserved minor-groove DNA binding loop. In Epstein-Barr Virus [HHV-4] UNG, this motif forms a disc-shaped loop structure of unclear significance. To ascertain the biological role of the loop insertion, we determined the crystal structure of Kaposi's sarcoma-associated herpesvirus [HHV-8] UNG (kUNG) in its product complex with a uracil-containing dsDNA, as well as two structures of kUNG in its apo state. We find the disc-like conformation is conserved, but only when the kUNG DNA-binding cleft is occupied. Surprisingly, kUNG uses this structure to flip the orphaned partner base of the substrate deoxyuridine out of the DNA duplex while retaining canonical UNG deoxyuridine-flipping and catalysis. The orphan base is stably posed in the DNA major groove which, due to DNA backbone manipulation by kUNG, is more open than in other UNG-dsDNA structures. Mutagenesis suggests a model in which the kUNG loop is pinned outside the DNA-binding cleft until DNA docking promotes rigid structuring of the loop and duplex nucleotide flipping, a novel observation for UNGs.


Growth phase and SpvR regulation of transcription of Salmonella typhimurium spvABC virulence genes.
The 90 kb virulence plasmid of Salmonella typhimurium is required for bacterial growth beyond the small intestine to deeper tissues such as the spleen and liver of orally inoculated mice. We constructed transcriptional lacZ fusions within the cloned plasmid-borne virulence genes spvA, spvB and spvC of S. typhimurium to demonstrate that spvR encodes a trans-acting positive regulator for the transcription of spvA, spvB and spvC. Data suggesting that the activation of spvABC transcription is dependent on the growth phase of both S. typhimurium and Escherichia coli grown in Luria Broth (LB) are also presented. Complementation experiments for virulence in mice confirmed that at least spvR and spvC are virulence genes and further suggested that the spvRABC gene cluster consists of at least three transcriptional units containing spvR, spvC and spvABC, respectively. Reinitiation of transcription at spvC was confirmed in vitro, using a lacZ fusion, and was shown to be independent of SpvR-mediated control in LB.


Predicting Health Care Cost Transitions Using a Multidimensional Adaptive Prediction Process.
Managing population health requires meeting individual care needs while striving for increased efficiency and quality of care. Predictive models can integrate diverse data to provide objective assessment of individual prospective risk to identify individuals requiring more intensive health management in the present. The purpose of this research was to develop and test a predictive modeling approach, Multidimensional Adaptive Prediction Process (MAPP). MAPP is predicated on dividing the population into cost cohorts and then utilizing a collection of models and covariates to optimize future cost prediction for individuals in each cohort. MAPP was tested on 3 years of administrative health care claims starting in 2009 for health plan members (average n=25,143) with evidence of coronary heart disease. A "status quo" reference modeling methodology applied to the total annual population was established for comparative purposes. Results showed that members identified by MAPP contributed $7.9 million and $9.7 million more in 2011 health care costs than the reference model for cohorts increasing in cost or remaining high cost, respectively. Across all cohorts, the additional accurate cost capture of MAPP translated to an annual difference of $1882 per member, a 21% improvement, relative to the reference model. The results demonstrate that improved future cost prediction is achievable using a novel adaptive multiple model approach. Through accurate prospective identification of individuals whose costs are expected to increase, MAPP can help health care entities achieve efficient resource allocation while improving care quality for emergent need individuals who are intermixed among a diverse set of health care consumers.


A feasibility study examining the effect on lung cancer diagnosis of offering a chest X-ray to higher-risk patients with chest symptoms: protocol for a randomized controlled trial.
In order to improve lung cancer survival in the UK, a greater proportion of resectable cancers must be diagnosed. It is likely that resectability rates would be increased by more timely diagnosis. Aside from screening, the only way of achieving this is to reduce the time to diagnosis in symptomatic cancers. Currently, lung cancers are mainly diagnosed by general practitioners (GPs) using the National Institute for Health and Clinical Excellence (NICE) guidelines for urgent referral for chest X-ray, which recommend urgent imaging or referral for patients who have one of a number of chest symptoms for more than 3 weeks. We are proposing to expand this recommendation to include one of a number of chest symptoms of any duration in higher-risk patients. We intend to conduct a trial of imaging in these higher-risk patients and compare it with NICE guidelines to see if imaging improves stage at diagnosis and resection rates. This trial would have to be large (and consequently resource-intensive) because most of these patients will not have lung cancer, making optimal design crucial. We are therefore conducting a pilot trial that will ascertain the feasibility of running a full trial and provide key information that will be required in order to design the full trial. This trial will assess the feasibility and inform the design of a large, UK-wide, clinical trial of a change to the NICE guidelines for urgent referral for chest X-ray for suspected lung cancer. It utilizes a combination of workshop, health economic, quality of life, qualitative, and quantitative methods in order to fully assess feasibility. Clinicaltrials.gov NCT01344005.


Severity assessment for lower respiratory tract infections: potential use and validity of the CRB-65 in primary care.
To explore the potential use of the CRB-65 rule (based on Confusion, Respiratory rate, Blood pressure and age >65 years) in adults with lower respiratory tract infection (LRTI) in primary care. Primary care clinicians in 13 European countries recorded antibiotic treatment and clinical features for adults with LRTI. Patients recorded daily symptoms. Multilevel regression models determined the association between an elevated CRB-65 score and prolonged moderately severe symptoms, hospitalisation, and time to recovery. Sensitivity analyses used zero imputation. Respiratory rate and blood pressure were recorded in 22.7% and 31.9% of patients, respectively. A total of 2,690 patients completed symptom diaries. The CRB-65 could be calculated for 339 (12.6%). A score of >1 was not significantly associated with prolonged moderately severe symptoms (odds ratio (OR) 0.42, 95% CI 0.04 to 4.19) or hospitalisations (OR 3.12, 95% CI 0.16 to 60.24), but was associated with prolonged time to self-reported recovery when using zero imputation (hazard ratio (HR) 0.75, 95% CI 0.64 to 0.88). Respiratory rate and blood pressure are infrequently measured in adults with LRTI. We found no evidence to support using the CRB-65 rule in the assessment of LRTI in primary care. However, it is unclear whether it is of value if used only in patients where the primary care clinician suspects pneumonia.


Functional anatomic models of language: assembling the pieces.
In the past few years, a series of influential review articles have summarized the state of the art with respect to cortical models of language organization. The present article is a mini-review and conceptual meta-analysis of several of the most prominent recent contributions. Based on the models, the authors extract some generalizations to arrive at a more robust model that 1) does justice to the range of neurological data, 2) is more connected to research in linguistics and psycholinguistics, and 3) stimulates hypothesis-driven research in this domain. In particular, the article attempts to unify a few of the current large-scale models of the functional neuroanatomy of language in a more principled manner. First, the authors argue that the relevant type of processing in a given cortical area, that is, memorizing (temporal cortex) versus analyzing (parietal) versus synthesizing (frontal), lies at the basis of local neuronal structure and function. Second, from an anatomic perspective, more dorsal regions within each of these (temporal, parietal, and frontal) systems specialize more in phonological processing, middle areas in syntactic processing, and more ventral areas in semantic processing.


Inhibition of N-type Ca2+ channels ameliorates an imbalance in cardiac autonomic nerve activity and prevents lethal arrhythmias in mice with heart failure.
Dysregulation of autonomic nervous system activity can trigger ventricular arrhythmias and sudden death in patients with heart failure. N-type Ca(2+) channels (NCCs) play an important role in sympathetic nervous system activation by regulating the calcium entry that triggers release of neurotransmitters from peripheral sympathetic nerve terminals. We have investigated the ability of NCC blockade to prevent lethal arrhythmias associated with heart failure. We compared the effects of cilnidipine, a dual N- and L-type Ca(2+) channel blocker, with those of nitrendipine, a selective L-type Ca(2+) channel blocker, in transgenic mice expressing a cardiac-specific, dominant-negative form of neuron-restrictive silencer factor (dnNRSF-Tg). In this mouse model of dilated cardiomyopathy leading to sudden arrhythmic death, cardiac structure and function did not significantly differ among the control, cilnidipine, and nitrendipine groups. However, cilnidipine dramatically reduced arrhythmias in dnNRSF-Tg mice, significantly improving their survival rate and correcting the imbalance between cardiac sympathetic and parasympathetic nervous system activity. A β-blocker, bisoprolol, showed similar effects in these mice. Genetic titration of NCCs, achieved by crossing dnNRSF-Tg mice with mice lacking CACNA1B, which encodes the α1 subunit of NCCs, improved the survival rate. With restoration of cardiac autonomic balance, dnNRSF-Tg;CACNA1B(+/-) mice showed fewer malignant arrhythmias than dnNRSF-Tg;CACNA1B(+/+) mice. Both pharmacological blockade of NCCs and their genetic titration improved cardiac autonomic balance and prevented lethal arrhythmias in a mouse model of dilated cardiomyopathy and sudden arrhythmic death. Our findings suggest that NCC blockade is a potentially useful approach to preventing sudden death in patients with heart failure.


Effects of amphibian chytrid fungus exposure on American toads in the presence of an insecticide.
Abiotic factors such as pesticides may alter the impact of a pathogen on hosts, which could have implications for host-pathogen interactions and may explain variation in disease outbreaks in nature. In the present laboratory experiment, American toad (Anaxyrus americanus) metamorphs were exposed to the amphibian chytrid fungal pathogen Batrachochytrium dendrobatidis (Bd) and environmentally relevant concentrations of the insecticide malathion to determine whether malathion altered the effects of Bd exposure on growth and survival of toad metamorphs. Exposure to Bd significantly decreased survival over the 51 d of the experiment, suggesting that Bd could reduce recruitment into the terrestrial life stage when exposure occurs at metamorphosis. Malathion did not impact survival, but a 12-h exposure at metamorphosis significantly reduced terrestrial growth. Toads that were exposed to both Bd and malathion showed a nonsignificant trend toward the smallest growth compared with other treatments. The present study suggests that Bd may pose a threat to American toads even though population declines have not been observed for this species; in addition, the presence of both the insecticide malathion and Bd could reduce terrestrial growth, which could have implications for lifetime fitness and suggests that environmental factors could play a role in pathogen impacts in nature.


Prevention of ocular scarring after glaucoma filtering surgery using the monoclonal antibody LT1009 (Sonepcizumab) in a rabbit model.
Excessive scarring leading to failure of the filtering bleb continues to be a major problem after glaucoma filtration surgery. This study examines the antifibrotic effects of the anti-S1P monoclonal antibody LT1009 (Sonepcizumab) in prolonging bleb survival in a rabbit model of glaucoma filtering surgery. The frequency of LT1009 dosage was determined initially using an enzyme-linked immunosorbent assay assay measuring LT1009 eye tissue retention in 6 New Zealand White rabbits. A further 21 New Zealand White rabbits underwent glaucoma filtering surgery. Bleb tissues were observed and compared clinically and histologically. The duration of bleb elevation was compared among LT1009, balanced saline solution (BSS) negative control, and mitomycin-C (MMC)-positive control. The mean duration of bleb survival was 28.5±8.5 days for rabbits receiving injections of LT1009, 21.0±5.6 days for those receiving injections of BSS, and 33.8±5.6 days for rabbits receiving MMC. Analysis of variance with post hoc testing suggests a statistically significant trend of improvement in bleb duration for LT1009 when compared with BSS controls. Nonpainful, upper eyelid edema was noted after 5 injections of LT1009, which resolved over a 10-day period. MMC eyes developed avascular conjunctivas with areas of thinning and sparse cellularity, whereas the conjunctiva of LT1009 and BSS eyes remained relatively normal. The monoclonal antibody LT1009 demonstrated a longer duration of bleb elevation than BSS control without adverse conjunctival effects associated with MMC. However, after multiple doses LT1009 use was associated with short-term upper eyelid edema.


A classification framework for Bacillus anthracis defined by global genomic structure.
Bacillus anthracis, the causative agent of anthrax, is a considerable global health threat affecting wildlife, livestock, and the general public. In this study, whole-genome sequence analysis of over 350 B. anthracis isolates was used to establish a new high-resolution global genotyping framework that is both biogeographically informative and compatible with multiple genomic assays. The data presented in this study shed new light on the diverse global dissemination of this species and indicate that many lineages may be uniquely suited to the geographic regions in which they are found. In addition, we demonstrate that plasmid genomic structure for this species is largely consistent with chromosomal population structure, suggesting vertical inheritance in this bacterium has contributed to its evolutionary persistence. This classification methodology is the first based on population genomic structure for this species and has potential use for local and broader institutions seeking to understand both disease outbreak origins and recent introductions. In addition, we provide access to a newly developed genotyping script as well as the full whole-genome sequence analyses output for this study, allowing future studies to rapidly employ and append their data in the context of this global collection. This framework may act as a powerful tool for public health agencies, wildlife disease laboratories, and researchers seeking to utilize and expand this classification scheme for further investigations into B. anthracis evolution.


Optimizing a community-engaged multi-level group intervention to reduce substance use: an application of the multiphase optimization strategy.
Rates of alcohol and illicit drug use (AIDU) are consistently similar across racial groups (Windsor and Negi, J Addict Dis 28:258-68, 2009; Keyes et al. Soc Sci Med 124:132-41, 2015). Yet AIDU has significantly higher consequences for residents in distressed communities with concentrations of African Americans (DCAA - i.e., localities with high rates of poverty and crime) who also have considerably less access to effective treatment of substance use disorders (SUD). This project is optimizing Community Wise, an innovative multi-level behavioral-health intervention created in partnership with service providers and residents of distressed communities with histories of SUD and incarceration, to reduce health inequalities related to AIDU. Grounded in critical consciousness theory, community-based participatory research principles (CBPR), and the multiphase optimization strategy (MOST), this study employs a 2 × 2 × 2 × 2 factorial design to engineer the most efficient, effective, and scalable version of Community Wise that can be delivered for US$250 per person or less. This study is fully powered to detect change in AIDU in a sample of 528 men with a histories of SUD and incarceration, residing in Newark, NJ in the United States. A community collaborative board oversees recruitment using a variety of strategies including indigenous field worker sampling, facility-based sampling, community advertisement through fliers, and street outreach. Participants are randomly assigned to one of 16 conditions that include a combination of the following candidate intervention components: peer or licensed facilitator, group dialogue, personal goal development, and community organizing. All participants receive a core critical-thinking component. Data are collected at baseline plus five post-baseline monthly follow ups. Once the optimized Community Wise intervention is identified, it will be evaluated against an existing standard of care in a future randomized clinical trial. This paper describes the protocol of the first ever study using CBPR and MOST to optimize a substance use intervention targeting a marginalized population. Data from this study will culminate in an optimized Community Wise manual; enhanced methodological strategies to develop multi-component scalable interventions using MOST and CBPR; and a better understanding of the application of critical consciousness theory to the field of health inequalities related to AIDU. ClinicalTrials.gov, NCT02951455 . Registered on 1 November 2016.


How do species, population and active ingredient influence insecticide susceptibility in Culicoides biting midges (Diptera: Ceratopogonidae) of veterinary importance?
Culicoides biting midges are biological vectors of internationally important arboviruses of livestock and equines. Insecticides are often employed against Culicoides as a part of vector control measures, but systematic assessments of their efficacy have rarely been attempted. The objective of the present study is to determine baseline susceptibility of multiple Culicoides vector species and populations in Europe and Africa to the most commonly used insecticide active ingredients. Six active ingredients are tested: three that are based on synthetic pyrethroids (alpha-cypermethrin, deltamethrin and permethrin) and three on organophosphates (phoxim, diazinon and chlorpyrifos-methyl). Susceptibility tests were conducted on 29,064 field-collected individuals of Culicoides obsoletus Meigen, Culicoides imicola Kieffer and a laboratory-reared Culicoides nubeculosus Meigen strain using a modified World Health Organization assay. Populations of Culicoides were tested from seven locations in four different countries (France, Spain, Senegal and South Africa) and at least four concentrations of laboratory grade active ingredients were assessed for each population. The study revealed that insecticide susceptibility varied at both a species and population level, but that broad conclusions could be drawn regarding the efficacy of active ingredients. Synthetic pyrethroid insecticides were found to inflict greater mortality than organophosphate active ingredients and the colony strain of C. nubeculosus was significantly more susceptible than field populations. Among the synthetic pyrethroids, deltamethrin was found to be the most toxic active ingredient for all species and populations. The data presented represent the first parallel and systematic assessment of Culicoides insecticide susceptibility across several countries. As such, they are an important baseline reference to monitor the susceptibility status of Culicoides to current insecticides and also to assess the toxicity of new active ingredients with practical implications for vector control strategies.


Training recollection in healthy older adults: clear improvements on the training task, but little evidence of transfer.
Normal aging holds negative consequences for memory, in particular for the ability to recollect the precise details of an experience. With this in mind, Jennings and Jacoby (2003) developed a recollection training method using a single-probe recognition memory paradigm in which new items (i.e., foils) were repeated during the test phase at increasingly long intervals. In previous reports, this method has appeared to improve older adults' performance on several non-trained cognitive tasks. We aimed to further examine potential transfer effects of this training paradigm and to determine which cognitive functions might predict training gains. Fifty-one older adults were assigned to either recollection training (n = 30) or an active control condition (n = 21) for six sessions over 2 weeks. Afterward, the recollection training group showed a greatly enhanced ability to reject the repeated foils. Surprisingly, however, the training and the control groups improved to the same degree in recognition accuracy (d') on their respective training tasks. Further, despite the recollection group's significant improvement in rejecting the repeated foils, we observed little evidence of transfer to non-trained tasks (including a temporal source memory test). Younger age and higher baseline scores on a measure of global cognitive function (as measured by the Montreal Cognitive Assessment tool) and working memory (as measured by Digit Span Backward) predicted gains made by the recollection training group members.


Human salivary acidic proline-rich proteins and statherin promote the attachment of Actinomyces viscosus LY7 to apatitic surfaces.
Actinomyces viscosus LY7 cells adsorbed in high numbers to experimental pellicles formed on hydroxyapatite (HA) from human parotid or submandibular saliva but not to pellicles prepared from human plasma or serum. To determine the nature of the salivary components responsible for promoting adhesion, pellicles were prepared from fractions of submandibular and parotid saliva obtained by chromatography on Trisacryl GF 2000 columns. Adsorption of LY7 cells was promoted by two groups of fractions. Each group was rechromatographed on DEAE-agarose. Fractions which promoted adsorption of LY7 cells were found by polyacrylamide gel electrophoresis to contain the acidic proline-rich proteins (PRPs) and statherin. Pellicles prepared from 12-micrograms/ml solutions of pure PRP-1, PRP-2, or parotid isoelectric focusing (PIF-slow) variant promoted maximal adsorption of A. viscosus LY7 cells. Somewhat higher concentrations of PRP-3 and PRP-4 were required for maximal adsorption, indicating that the 44-residue carboxy-terminal segment of PRP-1, PRP-2, and PIF-slow enhances LY7 binding but is not essential. Much higher concentrations of statherin were required to promote LY7 adsorption. Adsorption of LY7 cells to pellicles prepared from PRP-1 was not affected over the range of pH 5 to 8. Adsorption was also not inhibited by 50 mM lactose, which is consistent with the notion that type 1 fimbriae, rather than type 2 fimbriae, were responsible. A. viscosus T14, Actinomyces odontolyticus ATCC 17982, and Actinomyces israelii 12597 also adsorbed to PRP-1 pellicles, whereas Actinomyces naeslundii ATCC 12104 did not. Although A. viscosus cells bind strongly to adsorbed PRP-1, the presence of PRP-1 or PRP-3 in solution did not inhibit adhesion. Similarly, [3H]PRP-1 did not bind to LY7 cells, nor was it degraded when incubated with the organism. However, LY7 cells adsorbed to [3H]PRP-1 pellicles. These data suggest that hidden molecular segments of PRP become exposed when the protein adsorbs to HA; these segments then react with adhesins of LY7 cells. The apparent ability of A. viscosus cells to recognize segments of PRPs which are exposed only in surface-adsorbed molecules provides a novel mechanism which enables the organism to attach to teeth when suspended in salivary secretions.


[Research of anti-aging mechanism of ginsenoside Rg1 on brain].
Neurodegenerative disease is common and frequently occurs in elderly patients. Previous studies have shown that ginsenoside Rg1 was able to inhibit senescent of brain, but the mechanism on the brain during the treatment remains elucidated. To study the mechanism of ginsenoside Rg1 in the process of anti-aging of brain, forty male SD rats were randomly divided into normal group, Rg1 normal group, brain aging model group and Rg1 brain aging model group, each group with 10 rats (brain aging model group: subcutaneous injection of D-galactose (120 mg kg(-1)), qd for 42 consecutive days; Rg1 brain aging model group: while copying the same test as that of brain aging model group, begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Rg1 normal group: subcutaneous injection of the same amount of saline; begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Normal: injected with an equal volume of saline within the same time. Perform the related experiment on the second day after finishing copying the model or the completion of the first two days of drug injections). Learning and memory abilities were measured by Morris water maze. The number of senescent cells was detected by SA-beta-Gal staining while the level of IL-1 and IL-6 proinflammatory cytokines in hippocampus were detected by ELISA. The activities of SOD, contents of GSH in hippo- campus were quantified by chromatometry. The change of telomerase activities and telomerase length were performed by TRAP-PCR and southern blotting assay, respectively. It is pointed that, in brain aging model group, the spatial learning and memory capacities were weaken, SA-beta-Gal positive granules increased in section of brain tissue, the activity of antioxidant enzyme SOD and the contents of GSH decreased in hippocampus, the level of IL-1 and IL-6 increased in hippocampus, while the length of telomere and the activity of telomerase decreased in hippocampus. Rats of Rg1 brain aging group had their spatial learning and memory capacities enhanced, SA-beta-Gal positive granules in section of brain tissue decreased, the activity of antioxidant enzyme SOD and the contents of GSH increased in hippocampus, the level of IL-1 and IL-6 in hippocampus decreased, the length contraction of telomere suppressed while the change of telomerase activity increased in hippocampus. Compared with that of normal group, the spatial learning and memory capacities were enhanced in Rg1 normal group, SA-beta-Gal positive granules in section of brain tissue decreased in Rg1 normal group, the level of IL-1 and IL-6 in hippocampus decreased in Rg1 normal group. The results indicated that improvement of antioxidant ability, regulating the level of proinflammatory cytokines and regulation of telomerase system may be the underlying anti-aging mechanism of Ginsenoside Rg1.


[An approach to prevent lifestyle-related diseases of children in collaboration with various organizations in Tokushima].
To support the goal of "Lifetime health promotion from childhood", a Committee for Strategies to Prevent Lifestyle-related Diseases was established as part of the Tokushima Prefecture Medical Association in 2000. In this report, we present the activities of this committee, in collaboration with various organizations such as schools, a medical association, health administrators and universities. In 2000, a physical survey was performed for all students in primary and junior high schools in Tokushima prefecture. Subsequently, a software program for determining the degree of obesity using the standard body weight of Tokushima children was produced. In 2001, the committee conducted a survey concerning measures taken against lifestyle-related diseases by each organization. In 2003, a "Health management system for obesity in children" and a "School urine examination system" were established to identify high-risk students who should be taken to consult primary physicians. These medical intervention systems have allowed continuous calculation of real numbers and actual status of problems with overweight and obese children. Moreover, we performed lifestyle habit surveys among about 3000 students and produced manuals for population-based approaches. Compared with nationwide values, there was no difference in height, but the weight and BMI (Body Mass Index) of Tokushima students were larger. The survey concerning measures against lifestyle-related diseases clarified the present status of school health committees, staffing available to provide individual/nutritional guidance and the execution rate of collaborative projects in each organization. The intervention systems for visits to primary physicians have continued to show almost constant consultation rates. Approximately 80% of severely obese children had at least one medical problem. The lifestyle habits survey did not identify any marked differences in children of Tokushima Prefecture compared with nationwide values, except for a slightly earlier waking-up time. However, this survey demonstrated differences in lifestyle habits according to the body physique, and a relationship between eating meals with the family and other lifestyle habits. The numbers of overweight and severely obese children in Tokushima have been decreasing since the peaks of 2001 and 2002. Activities to prevent lifestyle-related diseases from childhood have continued in collaboration with various organizations in Tokushima. The prefecture-wide physical surveys and high-risk intervention strategies might have had good social effects in Tokushima. As a result, the number of obese children may be decreasing.


Lack of adequate sun protection for children with oculocutaneous albinism in South Africa.
Childhood is a high risk time for ultraviolet induced skin damage as this age group has more time and opportunity to be outdoors in the sun. Children in Africa with the inherited condition oculocutaneous albinism (OCA) are especially vulnerable due to their lack of protective melanin. They are highly susceptible to developing skin lesions that have both cosmetic and health complications, with a high risk of developing skin cancers. The study aimed to explore the adequacy of sun protection strategies of children with albinism in order to inform future provision. Community based participatory research methods were employed to investigate sun protection strategies in 90 pupils with OCA (40 female and 50 male) boarding at a special school educating pupils with visual impairment in a rural area of northern South Africa. Hats worn and sunscreen preparations used were examined during semi-structured face to face interviews conducted in small peer groups. The resident nurse interpreted if necessary and provided additional information on monitoring and treatment of skin lesions. Participants with albinism in this study were exposed to high levels of ultraviolet radiation throughout the year and showed skin damage despite wearing protective head gear. All except one pupil possessed at least one hat, with a mean brim width of 5.4 cm. Gender differences in sun avoidance behaviour were documented, with females seeking shade during recreational periods and males playing soccer outside. Although 38% of pupils were using a sunscreen with a sun protection factor (SPF) rating, only 12% had government sponsored tubes of SPF15 cream. Government sponsored sunscreen preparations were only provided if actively sought, involving time consuming trips to regional hospitals, with inadequate availability and insufficient supply. Children with albinism living away from home in rural areas appear to have inadequate sun protection strategies. Changes in health policy could address these deficiencies. We recommend providing more detailed health care information, giving advice on appropriate styles of hat to wear and how to assess commercial SPF products. Health promotional material should also be evaluated to determine its effectiveness among user groups.


Moving beyond Mindfulness: Defining Equanimity as an Outcome Measure in Meditation and Contemplative Research.
In light of a growing interest in contemplative practices such as meditation, the emerging field of contemplative science has been challenged to describe and objectively measure how these practices affect health and well-being. While "mindfulness" itself has been proposed as a measurable outcome of contemplative practices, this concept encompasses multiple components, some of which, as we review here, may be better characterized as equanimity. Equanimity can be defined as an even-minded mental state or dispositional tendency toward all experiences or objects, regardless of their origin or their affective valence (pleasant, unpleasant, or neutral). In this article we propose that equanimity be used as an outcome measure in contemplative research. We first define and discuss the inter-relationship between mindfulness and equanimity from the perspectives of both classical Buddhism and modern psychology and present existing meditation techniques for cultivating equanimity. We then review psychological, physiological, and neuroimaging methods that have been used to assess equanimity, either directly or indirectly. In conclusion, we propose that equanimity captures potentially the most important psychological element in the improvement of well-being, and therefore should be a focus in future research studies.


Malignant melanoma in the elderly: different regional disease and poorer prognosis.
Age is a poor prognostic factor in melanoma patients. Elderly melanoma patients have a different presentation and clinical course than younger patients. We evaluated the impact of age ≥70 years (yrs) on the diagnosis and natural history of melanoma. Retrospective review of 610 patients with malignant melanoma entered into a prospective sentinel lymph node (SLN) database, treated from June 1997 to June 2010. Disease characteristics and clinical outcomes were compared between patients ≥70 yrs vs. <70 yrs of age. 237 patients (39%) were ≥70 yrs. Elderly patients had a higher proportion of head and neck melanomas (34% vs. 20%, p<0.001), and greater mean tumor thickness (2.4mm vs. 1.8mm, p<0.001). A greater proportion of T3 or T4 melanoma was seen in the elderly (p<0.001) as well as a greater mean number of mitotic figures: 3.6/mm(2) vs. 2.7/mm(2) (p=0.005). Despite greater mean thickness, the incidence of SLN metastases was less in the ≥70 yrs group with T3/T4 melanomas (18% vs. 33%, p=0.02). The elderly had a higher rate of local and in-transit recurrences, 14.5% vs. 3.4% at 5 yrs (p<0.001). 5 yr disease-specific mortality and overall mortality were worse for those ≥70 yrs: 16% vs. 8% (p=0.004), and 30% vs. 12% (p<0.001), respectively. Elderly (≥70 yrs) melanoma patients present with thicker melanomas and a higher mitotic rate but have fewer SLN metastases. Melanoma in the elderly is more common on the head and neck. Higher incidence of local/in-transit metastases is seen among the elderly. Five-year disease-specific mortality and overall mortality are both worse for these patients.


Origin, Reactivity, and Bioavailability of Mercury in Wildfire Ash.
Wildfires are expected to become more frequent and intensive at the global scale due to climate change. Many studies have focused on the loss of mercury (Hg) from burned forests; however, little is known about the origins, concentration, reactivity, and bioavailability of Hg in residual ash materials in postfire landscapes. We examine Hg levels and reactivity in black ash (BA, low burn intensity) and white ash (WA, high burn intensity) generated from two recent northern California wildfires and document that all ash samples contained measurable, but highly variable, Hg levels ranging from 4 to 125 ng/g dry wt. ( n = 28). Stable Hg isotopic compositions measured in select ash samples suggest that most Hg in wildfire ash is derived from vegetation. Ash samples had a highly variable fraction of Hg in recalcitrant forms (0-75%), and this recalcitrant Hg pool appears to be associated with the black carbon fraction in ash. Both BA and WA were found to strongly sequester aqueous inorganic Hg but not gaseous elemental Hg under controlled conditions. During anoxic ash incubation with natural surface water, we find that Hg in most ash samples had a minimal release and low methylation potential. Thus, the formation of wildfire ash can sequester Hg into relatively nonbioavailable forms, attenuating the potentially adverse effects of Hg erosion and transport to aquatic environments along with eroded wildfire ash.


DNA hypermethylation biomarkers to predict response to cisplatin treatment, radiotherapy or chemoradiation: the present state of art.
Concurrent platinum-based chemoradiation significantly improved survival of advanced stage cervical patients over radiotherapy alone. However, the 5-year overall survival is still only 66%. Presently, no biomarkers are available to select those cervical cancer patients that might benefit from concurrent platinum-based chemoradiation therapy. DNA methylation is a well-established contributor to the regulation of gene transcription, predominantly causing transcriptional silencing. Differences in promoter hypermethylation patterns and subsequent silencing, could contribute to the variety of responses observed in clinical practice. Several clinical trials on various malignancies reported a better response when Decitabine was administered prior to or in combination with standard therapy. This sensitization is thought to be due to re-expression of tumor suppressor genes. However, not all patients might benefit from demethylating agents, since re-expression of oncogenes could render patients more resistant. In this review, we summarized the present state of art regarding hypermethylated genes and their affected signaling pathways that are associated with outcome after cisplatin treatment, radiotherapy or chemoradiation. Since only few studies were reported in cervical cancer, other malignancies were reviewed as well. From the data presented in this review, we conclude that, in order to select patients that benefit most optimally from demethylating strategies, a comprehensive screening of a large panel of methylation markers, associated with both good as well as poor clinical outcome have to be investigated. Since such panels are not available at this moment, global methylation screening approaches are required to profile such methylated genes. Such methylated gene profiles might be very useful to optimize personalized treatment planning not only in cervical cancer but also in other malignancies.


Challenging the in-vivo assessment of biomechanical properties of the uterine cervix: A critical analysis of ultrasound based quasi-static procedures.
Measuring the stiffness of the uterine cervix might be useful in the prediction of preterm delivery, a still unsolved health issue of global dimensions. Recently, a number of clinical studies have addressed this topic, proposing quantitative methods for the assessment of the mechanical properties of the cervix. Quasi-static elastography, maximum compressibility using ultrasound and aspiration tests have been applied for this purpose. The results obtained with the different methods seem to provide contradictory information about the physiologic development of cervical stiffness during pregnancy. Simulations and experiments were performed in order to rationalize the findings obtained with ultrasound based, quasi-static procedures. The experimental and computational results clearly illustrate that standardization of quasi-static elastography leads to repeatable strain values, but for different loading forces. Since force cannot be controlled, this current approach does not allow the distinction between a globally soft and stiff cervix. It is further shown that introducing a reference elastomer into the elastography measurement might overcome the problem of force standardization, but a careful mechanical analysis is required to obtain reliable stiffness values for cervical tissue. In contrast, the maximum compressibility procedure leads to a repeatable, semi-quantitative assessment of cervical consistency, due to the nonlinear nature of the mechanical behavior of cervical tissue. The evolution of cervical stiffness in pregnancy obtained with this procedure is in line with data from aspiration tests.


Extracellular FGF-1 acts as a lens differentiation factor in transgenic mice.
The vertebrate ocular lens undergoes a spatially defined pattern of differentiation which may be regulated by the ocular distribution of proteins from the fibroblast growth factor (FGF) family. The ability of altered FGF-1 (acidic FGF) distribution to disrupt the normal pattern of lens differentiation was evaluated by the production of transgenic mice which express FGF-1 under the control of the lens-specific alpha A-crystallin promoter. Since FGF-1 lacks a classical signal peptide consensus sequence, transgenic mice were also produced with a chimeric construct containing the signal peptide sequence of the FGF-4 gene fused in frame to the coding sequences of the FGF-1 cDNA in order to obtain extracellular expression of the transgene. The presence of transgenic mRNA and protein was confirmed by in situ hybridization, Western analysis and immunohistochemistry. The ocular histology of newborn and young adult transgenic mice expressing FGF-1 without a signal peptide appeared normal. In contrast, mice expressing secreted FGF-1 exhibited lens abnormalities including the elongation of anterior epithelial cells. Epithelial cell elongation was accompanied by expression of the fiber cell differentiation marker, beta-crystallin. These observations provide an in vivo demonstration that FGF-1 can induce anterior lens epithelial cells to express characteristics consistent with the onset of fiber cell differentiation. The transgenic induction of differentiation confirms that normal lens morphology reflects an asymmetric distribution of inductive factors within the eye.