Genetic relatedness among hepatitis A virus strains associated with food-borne outbreaks.
The genetic characterization of hepatitis A virus (HAV) strains is commonly accomplished by sequencing subgenomic regions, such as the VP1/P2B junction. HAV genome is not extensively variable, thus presenting opportunity for sharing sequences of subgenomic regions among genetically unrelated isolates. The degree of misrepresentation of phylogenetic relationships by subgenomic regions is especially important for tracking transmissions. Here, we analyzed whole-genome (WG) sequences of 101 HAV strains identified from 4 major multi-state, food-borne outbreaks of hepatitis A in the Unites States and from 14 non-outbreak-related HAV strains that shared identical VP1/P2B sequences with the outbreak strains. Although HAV strains with an identical VP1/P2B sequence were specific to each outbreak, WG were different, with genetic diversity reaching 0.31% (mean 0.09%). Evaluation of different subgenomic regions did not identify any other section of the HAV genome that could accurately represent phylogenetic relationships observed using WG sequences. The identification of 2-3 dominant HAV strains in 3 out of 4 outbreaks indicates contamination of the implicated food items with a heterogeneous HAV population. However, analysis of intra-host HAV variants from eight patients involved in one outbreak showed that only a single sequence variant established infection in each patient. Four non-outbreak strains were found closely related to strains from 2 outbreaks, whereas ten were genetically different from the outbreak strains. Thus, accurate tracking of HAV strains can be accomplished using HAV WG sequences, while short subgenomic regions are useful for identification of transmissions only among cases with known epidemiological association.


Symptoms and self-care following pancreaticoduodenectomy: Perspectives from patients and healthcare professionals - Foundation for an interactive ICT application.
Poor prognosis and a problematic recovery period after pancreaticoduodenectomy means that patients may benefit from early detection of symptoms and support for self-management. Interactive Information and Communication Technology tools can be used for this purpose, but the content needs to be relevant to patients as well as healthcare professionals. To facilitate development of the content of an application for this purpose, the aim of this study was to explore common symptoms and self-care in the first six months after pancreaticoduodenectomy, as identified by patients and healthcare professionals. Data were collected through individual interviews with patients (n = 14), along with two focus group interviews and one individual interview with healthcare professionals (n = 10). Data were analysed using qualitative content analysis. Common symptoms after surgery were those related to eating, bowel function and emotional wellbeing, along with fatigue and pain. Some self-care activities and advice were mentioned in the interviews. The patients often experienced a lack of advice on self-care at discharge. The results render knowledge of the symptoms it is important to be aware of and to assess regularly after pancreaticoduodenectomy. The results also contribute to knowledge about specific self-care related to these symptoms, even though it was not extensively described, and further research is needed to define evidence-based self-care advice.


Genotypic susceptibility testing of Mycobacterium tuberculosis isolates for amikacin and kanamycin resistance by use of a rapid sloppy molecular beacon-based assay identifies more cases of low-level drug resistance than phenotypic Lowenstein-Jensen testing.
Resistance to amikacin (AMK) and kanamycin (KAN) in clinical Mycobacterium tuberculosis strains is largely determined by specific mutations in the rrs gene and eis gene promoter. We developed a rapid, multiplexed sloppy molecular beacon (SMB) assay to identify these mutations and then evaluated assay performance on 603 clinical M. tuberculosis DNA samples collected in South Korea. Assay performance was compared to gold-standard phenotypic drug susceptibility tests, including Lowenstein-Jensen (LJ) absolute concentration, mycobacterial growth indicator tubes (MGIT), and TREK Sensititre MycoTB MIC plate (MycoTB) methods. Target amplicons were also tested for mutations by Sanger sequencing. The SMB assay correctly detected 115/116 mutant and mixed sequences and 487/487 wild-type sequences (sensitivity and specificity of 99.1 and 100%, respectively). Using the LJ method as the reference, sensitivity and specificity for AMK resistance were 92.2% and 100%, respectively, and sensitivity and specificity for KAN resistance were 87.7% and 95.6%, respectively. Mutations in the rrs gene were unequivocally associated with high-level cross-resistance to AMK and KAN in all three conventional drug susceptibility testing methods. However, eis promoter mutations were associated with KAN resistance using the MGIT or MycoTB methods but not the LJ method. No testing method associated eis promoter mutations with AMK resistance. Among the discordant samples with AMK and/or KAN resistance but wild-type sequence at the target genes, we discovered four new mutations in the whiB7 5' untranslated region (UTR) in 6/22 samples. All six samples were resistant only to KAN, suggesting the possible role of these whiB7 5' UTR mutations in KAN resistance.


Evaluation of cell-laden polyelectrolyte hydrogels incorporating poly(L-Lysine) for applications in cartilage tissue engineering.
To address the lack of reliable long-term solutions for cartilage injuries, strategies in tissue engineering are beginning to leverage developmental processes to spur tissue regeneration. This study focuses on the use of poly(L-lysine) (PLL), previously shown to up-regulate mesenchymal condensation during developmental skeletogenesis in vitro, as an early chondrogenic stimulant of mesenchymal stem cells (MSCs). We characterized the effect of PLL incorporation on the swelling and degradation of oligo(poly(ethylene) glycol) fumarate) (OPF)-based hydrogels as functions of PLL molecular weight and dosage. Furthermore, we investigated the effect of PLL incorporation on the chondrogenic gene expression of hydrogel-encapsulated MSCs. The incorporation of PLL resulted in early enhancements of type II collagen and aggrecan gene expression and type II/type I collagen expression ratios when compared to blank controls. The presentation of PLL to MSCs encapsulated in OPF hydrogels also enhanced N-cadherin gene expression under certain culture conditions, suggesting that PLL may induce the expression of condensation markers in synthetic hydrogel systems. In summary, PLL can function as an inductive factor that primes the cellular microenvironment for early chondrogenic gene expression but may require additional biochemical factors for the generation of fully functional chondrocytes.


Eculizumab Modifies Outcomes in Adults with Atypical Hemolytic Uremic Syndrome with Acute Kidney Injury.
Atypical hemolytic uremic syndrome (aHUS) is a rare disease associated with congenital or acquired genetic abnormalities that result in uncontrolled complement activation, leading to thrombotic microangiopathy and kidney failure. Until recently, the only treatment was plasma exchange or plasma infusion (PE/PI), but 60% of patients died or had permanent kidney damage despite treatment. Eculizumab, a complement inhibitor, has shown promising results in aHUS. However, data are mainly extracted from case reports or studies of heterogeneous cohorts, and no direct comparison with PE/PI is available. An observational retrospective study of adult, dialysis-dependent aHUS patients with acute kidney injury (AKI) who were treated with either PE/PI alone or with second-line eculizumab in our center. We compared the effect of PE/PI and eculizumab on kidney function, hypertension, proteinuria, hematologic values, relapse, and death. Thirty-one patients were included (females, 18; sporadic aHUS, 29; mean age, 46 ± 20 years). Twenty-six patients were treated with PE/PI alone, and 5 were deemed to be plasma-resistant and received eculizumab after stopping PE/PI. Among patients receiving eculizumab, 80% attained complete recovery of kidney function, 100% stopped dialysis, 20% had decreased proteinuria, and no patient relapsed (vs. 38.5, 50, 15.4, and 11.5%, respectively, of patients receiving only PE/PI). At 1-year of follow-up, no deaths had occurred in either group. Eculizumab shows greater efficacy than PE/PI alone for the treatment of adult aHUS patients with AKI. Prospective studies and meta-analyses are warranted to confirm our findings and set guidelines for treatment, monitoring, and maintenance.


Prognostic value of stage IV neuroblastoma metastatic immunophenotype in the bone marrow: preliminary report.
To correlate the immunophenotype of metastatic cells in the bone marrow of patients with neuroblastoma with early treatment failure. The studies were performed on bone marrow material obtained from children treated in the department of paediatric oncology, haematology, and transplantology, Poznan University of Medical Sciences, Poland from 1996 to 2003. Immunocytochemical analysis of nervous tissue markers (using the immunomax technique) was performed on 108 bone marrow preparations obtained from 36 children diagnosed with neuroblastoma (stage IV with bone marrow metastases). The analysis included expression of PGP 9.5 protein, substance P, chromogranin A, bombesin, galanin, neuropeptide Y (NPY), and vasoactive intestinal peptide in neuroblastoma metastatic cells defined by the expression of neurone specific enolase. Nineteen relapses occurred within 12 months of the end of treatment. Correlation between the various markers studied and early treatment failure, using Fisher's exact test, revealed that chromogranin A and NPY are strong indicators of an unfavourable prognosis in patients with stage IV neuroblastoma (p < 0.001 and p < 0.0002, respectively). Determination of metastatic cell immunophenotypes in bone marrow (particularly chromogranin A and NPY) may help establish the short term prognosis in children with neuroblastoma.


Multispecific lymphoid cell surface receptors.
When mice are sequentially immunized with two antigens to give an oligoclonal "double-binding" antibody response, there is a concomitant increase of "double-binding" cell surface receptors on their splenic lymphocytes. Competition studies suggest that the capacity to bind the two ligands, bovine pancreatic ribonuclease (EC 18.104.22.168) and a 2,4-dinitrophenyl (DNP) derivative, is a function of the same molecules. In ribo-nuclease-primed mice, an early response to bovine gamma globulin containing an average of 60 Dnp groups per molecule is the appearance of an increasing number of cells bearing surface receptors binding both ribonuclease and Dnp. Later, these double-binding cells are diluted by cells that bind Dnp, but not ribonuclease. The analogous phenomenon is observed when the two antigens are used in reverse order. While other reports suggest that there may be several different receptors in relatively undifferentiated cells from unimmunized mice, it seems likely that cells committed to antibody production carry a predominant multispecific cell surface immunoglobulin receptor.


Association of respiratory symptoms and lung function with occupation in the multinational Burden of Obstructive Lung Disease (BOLD) study.
Chronic obstructive pulmonary disease has been associated with exposures in the workplace. We aimed to assess the association of respiratory symptoms and lung function with occupation in the Burden of Obstructive Lung Disease study. We analysed cross-sectional data from 28 823 adults (≥40 years) in 34 countries. We considered 11 occupations and grouped them by likelihood of exposure to organic dusts, inorganic dusts and fumes. The association of chronic cough, chronic phlegm, wheeze, dyspnoea, forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV1)/FVC with occupation was assessed, per study site, using multivariable regression. These estimates were then meta-analysed. Sensitivity analyses explored differences between sexes and gross national income. Overall, working in settings with potentially high exposure to dusts or fumes was associated with respiratory symptoms but not lung function differences. The most common occupation was farming. Compared to people not working in any of the 11 considered occupations, those who were farmers for ≥20 years were more likely to have chronic cough (OR 1.52, 95% CI 1.19-1.94), wheeze (OR 1.37, 95% CI 1.16-1.63) and dyspnoea (OR 1.83, 95% CI 1.53-2.20), but not lower FVC (β=0.02 L, 95% CI -0.02-0.06 L) or lower FEV1/FVC (β=0.04%, 95% CI -0.49-0.58%). Some findings differed by sex and gross national income. At a population level, the occupational exposures considered in this study do not appear to be major determinants of differences in lung function, although they are associated with more respiratory symptoms. Because not all work settings were included in this study, respiratory surveillance should still be encouraged among high-risk dusty and fume job workers, especially in low- and middle-income countries.


Analysis of phi and chi 1 torsion angles for hen lysozyme in solution from 1H NMR spin-spin coupling constants.
Three-bond 3JHN alpha coupling constants have been determined for 106 residues and 3J alpha beta coupling constants have been measured for 57 residues of the 129-residue protein hen egg white lysozyme. These NMR data have been compared with torsion angles defined in the tetragonal and the triclinic crystal forms of the protein. For most residues the measured 3JHN alpha values were consistent with the phi torsion angles found in both crystal forms; the RMS difference between the coupling constants calculated by using the tetragonal crystal structure phi angles and the experimental 3JHN alpha values is 0.88 Hz. Thus there appears to be no significant averaging of the phi torsion angle either in the interior or at the surface of the protein. For 41 of the residues where 3J alpha beta coupling constants have been determined, the values are consistent with a single staggered conformation about the chi 1 torsion angle and there is complete agreement between the NMR data in solution and the torsion angles defined in the crystalline state. In contrast, for the other 16 residues where 3J alpha beta coupling constant values have been measured, the data indicate extensive motional averaging about the chi 1 torsion angle. These residues occur largely on the surface of the protein and examination of the crystal structures shows that many of these residues adopt a different conformation in the triclinic and tetragonal crystal forms and have high crystallographic temperature factors. It appears, however, that in solution conformational flexibility of the side chains of surface residues is significantly more pronounced than in individual crystal structures.


Case report: intra-tendinous ganglion of the anterior cruciate ligament in a young footballer.
A 20-year-old male medical student and keen rugby player presented with a 12-month history of progressively worsening right knee pain and stiffness with no history of trauma. Clinical examination revealed effusion and posterior knee pain exacerbated by end range movement and an extension lag of 15 degrees. Physiotherapy to improve the range of motion proved unsuccessful. Magnetic resonance imaging showed that the ACL was grossly thickened and displaced by material reported as mucoid in nature. There were also areas of focally high signal in relation to its tibial attachment and intra osseous small cysts. Arthroscopic examination revealed a ganglion related to the tibial attachment of the ACL and gross thickening and discoloration of the ACL. Biopsies were taken showing foci of mucoid degeneration in the ACL. A large intra-ACL mass of brownish coloured tissue was excised arthroscopically. Already at 2 weeks follow up the patient had greatly improved range of movement and was pain free. However, upon returning to rugby, joint instability was noticed and a tear of the ACL was confirmed. This rare clinical condition can be diagnosed with MRI and arthroscopic debridement effectively relieves symptoms. This case report illustrates that augmentation or reconstruction may end up being the definitive treatment for athletes. It may also offer some support to the argument that mucoid degeneration and ganglion cyst formation share a similar pathogenesis to intra-osseous cyst formation.


Prevascularized silicon membranes for the enhancement of transport to implanted medical devices.
Recent advances in drug delivery and sensing devices for in situ applications are limited by the diffusion-limiting foreign body response of fibrous encapsulation. In this study, we fabricated prevascularized synthetic device ports to help mitigate this limitation. Membranes with rectilinear arrays of square pores with widths ranging from 40 to 200 μm were created using materials (50 μm thick double-sided polished silicon) and processes (photolithography and directed reactive ion etching) common in the manufacturing of microfabricated sensors. Vascular endothelial cells responded to membrane geometry by either forming vascular tubes that extended through the pore or completely filling membrane pores after 4 days in culture. Although tube formation began to predominate overgrowth around 75 μm and continued to increase at even larger pore sizes, tubes formed at these large pore sizes were not completely round and had relatively thin walls. Thus, the optimum range of pore size for prevascularization of these membranes was estimated to be 75-100 μm. This study lays the foundation for creating a prevascularized port that can be used to reduce fibrous encapsulation and thus enhance diffusion to implanted medical devices and sensors. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1602-1609, 2016.


Shared decision making and risk communication in practice: a qualitative study of GPs' experiences.
Important barriers to the wider implementation of shared decision making remain. The experiences of professionals who are skilled in this approach may identify how to overcome these barriers. To identify the experiences and views of professionals skilled in shared decision making and risk communication, exploring the opportunities and challenges for implementation. Qualitative study. Gwent Health Authority. Exit interviews using focus group methodology with 20 GPs who had been in practice between 1 and 10 years, and participated in an explanatory trial lasting 6 months. The trial interventions comprised training in shared decision-making skills and the use of risk communication materials. The doctors consulted with up to 48 patients each (mean = 40, half of them audiotaped) for the study. The GPs indicated positive attitudes towards involving patients and described positive effects on their consultations. However, the frequency of applying the new skills and tools was limited outside the trial. Doctors were selective about when they felt greater patient involvement was appropriate and feasible, rather than seeking to apply the approaches to the majority of consultations. They felt they often responded to consumer preferences for low levels of involvement in decision making. Time limitations were important in not implementing the approach more widely. The promotion of 'patient involvement' appears likely to continue. Professionals appear receptive to this, and willing to acquire the relevant skills. Strategies for wider implementation of patient involvement could address how consultations are scheduled in primary care, and raise consumers' expectations or desires for involvement.


Modeling uncertainties in mining pillar stability analysis.
Many countries are now facing problems related to their past mining activities. One of the greatest problems concerns the potential surface instability. In areas where a room-and-pillar extraction method was used, deterministic methodologies are generally used to assess the hazard of surface collapses. However, those methodologies suffer from not being able to take into account all the uncertainties inherent in any hazard analysis. Through the practical example of the assessment of a single pillar stability in a very simple mining layout, this article introduces a logical framework that can be used to incorporate the different kinds of uncertainties related to data and models, as well as to specific expert's choices in the hazard or risk analysis process. Practical recommendations and efficient tools are also provided to help engineers and experts in their daily work.


Benefits and challenges with diagnosing chronic and late acute GVHD in children using the NIH consensus criteria.
Chronic graft-versus-host disease (cGVHD) and late acute graft-versus-host disease (L-aGVHD) are understudied complications of allogeneic hematopoietic stem cell transplantation in children. The National Institutes of Health Consensus Criteria (NIH-CC) were designed to improve the diagnostic accuracy of cGVHD and to better classify graft-versus-host disease (GVHD) syndromes but have not been validated in patients <18 years of age. The objectives of this prospective multi-institution study were to determine: (1) whether the NIH-CC could be used to diagnose pediatric cGVHD and whether the criteria operationalize well in a multi-institution study; (2) the frequency of cGVHD and L-aGVHD in children using the NIH-CC; and (3) the clinical features and risk factors for cGVHD and L-aGVHD using the NIH-CC. Twenty-seven transplant centers enrolled 302 patients <18 years of age before conditioning and prospectively followed them for 1 year posttransplant for development of cGVHD. Centers justified their cGVHD diagnosis according to the NIH-CC using central review and a study adjudication committee. A total of 28.2% of reported cGVHD cases was reclassified, usually as L-aGVHD, following study committee review. Similar incidence of cGVHD and L-aGVHD was found (21% and 24.7%, respectively). The most common organs involved with diagnostic or distinctive manifestations of cGVHD in children include the mouth, skin, eyes, and lungs. Importantly, the 2014 NIH-CC for bronchiolitis obliterans syndrome perform poorly in children. Past acute GVHD and peripheral blood grafts are major risk factors for cGVHD and L-aGVHD, with recipients ≥12 years of age being at risk for cGVHD. Applying the NIH-CC in pediatrics is feasible and reliable; however, further refinement of the criteria specifically for children is needed.


Application of spatio-temporal model to estimate burden of diseases, injuries and risk factors in Iran 1990 - 2013.
Identifying the burden of disease and its inequality between geographical regions is an important issue to study health priorities. Estimating burden of diseases using statistical models is inevitable especially in the context of rare data availability. To this purpose, the spatio-temporal model can provide a statistically sound approach for explaining the response variable observed over a region and various times. However, there are some methodological challenges in analysis of these complex data. Our primary objective is to provide some remedies to overcome these challenges. Data from nationally representative surveys and systematic reviews have been gathered across contiguous areal units over a period of more than 20 years (1990 - 2013). Generally, observations of areal units are spatially and temporally correlated in such a way that observations closer in space and time tend to be more correlated than observations farther away. It is critical to determine the correlation structure in space-time process which has been observed over a set of irregular regions. Moreover, these data sets are subject to high percentage of missing, including misaligned areal units, areas with small sample size, and may have nonlinear trends over space and time. Furthermore, the Gaussian assumption might be overly restrictive to represent the data. In this setting, the traditional statistical techniques are not appropriate and more flexible and comprehensive methodology is required. Particularly, we focus on approaches that allow extending spatio-temporal models proposed previously in the literature.Since statistical models include both continuous and categorical outcomes, we assume a latent variable framework for describing the underlying structure in mixed outcomes and use a conditionally autoregressive (CAR) prior for the random effects. In addition, we will employ misalignment modeling to combine incompatible areal units between data sources and/or over the years to obtain a unified clear picture of population health status over this period. In order to take parameter uncertainties into account, we pursue a Bayesian sampling-based inference. Hence, a hierarchical Bayes approach is constructed to model the data. The hierarchical structure enables us to "borrow information" from neighboring areal units to improve estimates for areas with missing values and small number of observations. For their general applicability and ease of implementation, the MCMC methods are the most adapted tool to perform Bayesian inference. This study aims to combine different available data sources and produce precise and reliable evidences for Iranian burden of diseases and risk factors and their disparities among geographical regions over time. Providing appropriate statistical methods and models for analyzing the data is undoubtedly crucial to circumvent the problems and obtain satisfactory estimates of model parameters and reach accurate assessment.


Liquid-liquid phase separation of type II diabetes-associated IAPP initiates hydrogelation and aggregation.
Amyloidoses (misfolded polypeptide accumulation) are among the most debilitating diseases our aging societies face. Amyloidogenesis can be catalyzed by hydrophobic-hydrophilic interfaces (e.g., air-water interface in vitro [AWI]). We recently demonstrated hydrogelation of the amyloidogenic type II diabetes-associated islet amyloid polypeptide (IAPP), a hydrophobic-hydrophilic interface-dependent process with complex kinetics. We demonstrate that human IAPP undergoes AWI-catalyzed liquid-liquid phase separation (LLPS), which initiates hydrogelation and aggregation. Insulin modulates these processes but does not prevent them. Using nonamyloidogenic rat IAPP, we show that, whereas LLPS does not require the amyloidogenic sequence, hydrogelation and aggregation do. Interestingly, both insulin and rat sequence delayed IAPP LLPS, which may reflect physiology. By developing an experimental setup and analysis tools, we show that, within the whole system (beyond the droplet stage), macroscopic interconnected aggregate clusters form, grow, fuse, and evolve via internal rearrangement, leading to overall hydrogelation. As the AWI-adsorbed gelled layer matures, its microviscosity increases. LLPS-driven aggregation may be a common amyloid feature and integral to pathology.


Reciprocity on Demand : Sharing and Exchanging Food in Northwestern Namibia.
Two competing models concerning food transfers prominent in the anthropological literature conceptualize such transfers either as sharing or as exchange. Sharing is understood as situational transactions formed through demands and unconditional giving, whereas reciprocal exchange is understood in terms of networking and keeping score. I propose that the picture is more complicated than these classifications suggests. Drawing on data collected in Northwestern Namibia, I show that sharing and reciprocal exchange are dynamically interrelated in actual food transfers. As a local norm, people can demand food from anyone, and they are typically given food in response to a demand. However, in practice, food transfer networks emerge (N = 62) that are highly reciprocal and fit the exchange model much better. Although the sharing norm makes no restrictions on whom to ask, in practice people often turn to their neighbors. Interpersonal dynamics account for why some of those ties become strongly reciprocal and others do not. Under these circumstances, unconditional sharing, a norm that has been viewed as an alternative to exchange, can lead to reciprocity via reciprocity on demand.


Geometry as a factor for tissue growth: towards shape optimization of tissue engineering scaffolds.
Scaffolds for tissue engineering are usually designed to support cell viability with large adhesion surfaces and high permeability to nutrients and oxygen. Recent experiments support the idea that, in addition to surface roughness, elasticity and chemistry, the macroscopic geometry of the substrate also contributes to control the kinetics of tissue deposition. In this study, a previously proposed model for the behavior of osteoblasts on curved surfaces is used to predict the growth of bone matrix tissue in pores of different shapes. These predictions are compared to in vitro experiments with MC3T3-E1 pre-osteoblast cells cultivated in two-millimeter thick hydroxyapatite plates containing prismatic pores with square- or cross-shaped sections. The amount and shape of the tissue formed in the pores measured by phase contrast microscopy confirms the predictions of the model. In cross-shaped pores, the initial overall tissue deposition is twice as fast as in square-shaped pores. These results suggest that the optimization of pore shapes may improve the speed of ingrowth of bone tissue into porous scaffolds.


Targeting of p32 in peritoneal carcinomatosis with intraperitoneal linTT1 peptide-guided pro-apoptotic nanoparticles.
Gastrointestinal and gynecological malignancies disseminate in the peritoneal cavity - a condition known as peritoneal carcinomatosis (PC). Intraperitoneal (IP) administration can be used to improve therapeutic index of anticancer drugs used for PC treatment. Activity of IP anticancer drugs can be further potentiated by encapsulation in nanocarriers and/or affinity targeting with tumor-specific affinity ligands, such as tumor homing peptides. Here we evaluated a novel tumor penetrating peptide, linTT1 (AKRGARSTA), as a PC targeting ligand for nanoparticles. We first demonstrated that the primary homing receptor for linTT1, p32 (or gC1qR), is expressed on the cell surface of peritoneal carcinoma cell lines of gastric (MKN-45P), ovarian (SKOV-3), and colon (CT-26) origin, and that peritoneal tumors in mice and clinical peritoneal carcinoma explants express p32 protein accessible from the IP space. Iron oxide nanoworms (NWs) functionalized with the linTT1 peptide were taken up and routed to mitochondria in cultured PC cells. NWs functionalized with linTT1 peptide in tandem with a pro-apoptotic [D(KLAKLAK)2] peptide showed p32-dependent cytotoxicity in MKN-45P, SKOV-3, and CT-26 cells. Upon IP administration in mice bearing MKN-45P, SKOV-3, and CT-26 tumors, linTT1-functionalized NWs showed robust homing and penetration into malignant lesions, whereas only a background accumulation was seen in control tissues. In tumors, the linTT1-NW accumulation was seen predominantly in CD31-positive blood vessels, in LYVE-1-positive lymphatic structures, and in CD11b-positive tumor macrophages. Experimental therapy of mice bearing peritoneal MKN-45P xenografts and CT-26 syngeneic tumors with IP linTT1-D(KLAKLAK)2-NWs resulted in significant reduction of weight of peritoneal tumors and significant decrease in the number of metastatic tumor nodules, whereas treatment with untargeted D(KLAKLAK)2-NWs had no effect. Our data show that targeting of p32 with linTT1 tumor-penetrating peptide improves tumor selectivity and antitumor efficacy of IP pro-apoptotic NWs. P32-directed intraperitoneal targeting of other anticancer agents and nanoparticles using peptides and other affinity ligands may represent a general strategy to increase their therapeutic index.


A cyanobacterial light activated adenylyl cyclase partially restores development of a Dictyostelium discoideum, adenylyl cyclase a null mutant.
A light-regulated adenylyl cyclase, mPAC, was previously identified from the cyanobacterium Microcoleus chthonoplastes PCC7420. MPAC consists of a flavin-based blue light-sensing LOV domain and a catalytic domain. In this work, we expressed mPAC in an adenylate cyclase A null mutant (aca-) of the eukaryote Dictyostelium discoideum and tested to what extent light activation of mPAC could restore the cAMP-dependent developmental programme of this organism. Amoebas of Dictyostelium, a well-established model organism, generate and respond to cAMP pulses, which cause them to aggregate and construct fruiting bodies. mPAC was expressed under control of a constitutive actin-15 promoter in D. discoideum and displayed low basal adenylyl cyclase activity in darkness that was about five-fold stimulated by blue light. mPAC expression in aca- cells marginally restored aggregation and fruiting body formation in darkness. However, more and larger fruiting bodies were formed when mPAC expressing cells were incubated in light. Extending former applications of light-regulated AC, these results demonstrate that mPAC can be used to manipulate multicellular development in eukaryotes in a light dependent manner.


SNAP: predict effect of non-synonymous polymorphisms on function.
Many genetic variations are single nucleotide polymorphisms (SNPs). Non-synonymous SNPs are 'neutral' if the resulting point-mutated protein is not functionally discernible from the wild type and 'non-neutral' otherwise. The ability to identify non-neutral substitutions could significantly aid targeting disease causing detrimental mutations, as well as SNPs that increase the fitness of particular phenotypes. Here, we introduced comprehensive data sets to assess the performance of methods that predict SNP effects. Along we introduced SNAP (screening for non-acceptable polymorphisms), a neural network-based method for the prediction of the functional effects of non-synonymous SNPs. SNAP needs only sequence information as input, but benefits from functional and structural annotations, if available. In a cross-validation test on over 80,000 mutants, SNAP identified 80% of the non-neutral substitutions at 77% accuracy and 76% of the neutral substitutions at 80% accuracy. This constituted an important improvement over other methods; the improvement rose to over ten percentage points for mutants for which existing methods disagreed. Possibly even more importantly SNAP introduced a well-calibrated measure for the reliability of each prediction. This measure will allow users to focus on the most accurate predictions and/or the most severe effects. Available at http://www.rostlab.org/services/SNAP.


Three-dimensional shape measurement of large-aperture aspheric mirrors by off-axis null Ronchi test.
An off-axis null Ronchi test is presented to measure the three-dimensional (3D) shape of a large-aperture aspheric mirror. The method designs curved fringe patterns as null sinusoidal gratings by means of phase information and ray tracing. In the process of measurement, the curved fringe patterns are displayed on a transmission-type liquid crystal display (T-LCD) screen, and a CCD camera records the fringe patterns containing the information of deviations of the mirror. The slopes of the deviations of the mirror are obtained by using the recorded fringe patterns. The deviations are restored by integrating, and then the 3D shape of the mirror can be reconstructed. Compared with the classical null Ronchi test, the method can provide enough measured data points and avoid the jagged edges of bands on the null gratings. Moreover, the method can conveniently change period and direction of the curved fringes and accurately control phase shifting. Computer simulations and a preliminary experiment are presented to show the performance of the method.


A New Surgical Technique for Closure of Pilonidal Sinus Defects: Triangular Closure Technique.
BACKGROUND We present a clinical experience with a new local flap procedure, namely the triangular closure technique, for reconstruction of sacrococcygeal skin defects resulting from excision of the pilonidal sinus. MATERIAL AND METHODS In this technique, the defect is surgically converted to a triangle in shape. Then, the triangular defect is closed by transposition of 2 skin flaps designed in an unequal z-plasty manner. Over 6 years, this technique has been used for closure of defects of chronic pilonidal sinus disease in 27 patients (6 females, 21 males). The size of the defect ranged between 3.5 cm and 12 cm in dimension. RESULTS A tension-free defect closure was obtained in all patients. All flaps except one healed with no complications. A mean follow-up of 3.62±1.77 months revealed aesthetically and functionally acceptable results with the obliteration of the natal cleft in all patients. No recurrence was observed in the follow-up period. CONCLUSIONS The triangular closure technique was found to be a useful technique for the treatment of pilonidal sinus with favorable results regarding the time before return to work.


Ultrastructural immunoperoxidase demonstration of autologous albumin in the alveolar capillary membrane and in the alveolar lining material in normal rats.
The location of autologous serum albumin within the alveolar-capillary membrane was studied in the rat under physiological conditions using antialbumin antibodies labeled with peroxidase. Albumin was detected in the lung interstitium, and in numerous pinocytic vesicles within endothelial cells and type I alveolar epithelial cells. The immunoreaction was also positive at the level of plasmalemmal membranes of both cell types and in the alveolar lining material.


Role stress among physician executives.
On the one hand, physician executives are clinicians who place value on professional autonomy. As clinicians, the best interests of the patient drive their decision making and their value system. On the other hand, as managers, physician executives serve as agents of an organization. Because of the differences in the two cultures, some physicians have called the physician executive position a "no man's land" To address these issues and answer the questions that surround them, the authors developed a survey that was mailed to a random sample of the membership of the American College of Physician Executives. Parts of the survey served in other studies of role conflict and role ambiguity. Parts of the survey are new, developed specifically to analyze the physician executive role. The findings are reported in this article.


El Niño-southern oscillation effect on a fire regime in northeastern Mexico has changed over time.
The El Niño Southern Oscillation (ENSO) is a climate-forcing mechanism that has been shown to affect precipitation and the occurrence of wildfires in many parts of the world. In the southern United States and northern Mexico, warm events (El Niño) are associated with moist winter conditions and fewer fires, while cool events (La Niñia) tend to favor dry winters and more fires. We tested this relationship in a region of northeastern Mexico by characterizing the historical fire regime and climatic influences: Fire regimes were reconstructed from fire-scar samples collected from 100 trees in three high-elevation sites on Peña Nevada in southern Nuevo Le6n. The sites were approximately 25 ha each, and the site centers were approximately 1 km apart. The earliest recorded fire occurred in 1521 and the time period we used for analysis was 1645-1929. The sites were characterized by frequent surface fires before the 1920s. In the three sites, mean fire intervals ranged from 8.6 to 9.6 years (all fires) and 11.9 to 18.6 years (fires that scarred > or = 25% of recording trees). The per-tree mean fire return interval was 17 years, and all three sites burned in the same year seven times between 1774 and 1929. After 1929, fires were nearly eliminated in all sites, likely due to human causes. We found a temporal change in the association between ENSO events and fires; before the 1830s La Niña events were significantly associated with fire years, while after the 1830s this association was not significant. In 1998, when the most severe El Niño event of the past century occurred, the three sites experienced severe, stand-replacing fires that killed many trees that had survived multiple surface fires in the past. Prior to the 1830s, fires tended to occur during dry La Niña years, but since then both La Niña and El Niño have been associated with dry years in this region, especially during the last three decades. This result suggests that ENSO effects have changed over time in this location and that phases of ENSO are not consistent indicators of precipitation, fire occurrence, or fire behavior in this area of northeastern Mexico.


Optical switching and logic gates with vertical cavity surface-emitting lasers.
We consider a cylindrical vertical cavity surface-emitting laser with a square-shaped active region, operating with the two Gauss-Hermite modes TEM(10) and TEM(01). Using a rate-equation model that includes the effects of the linewidth enhancement factor, we identify in the parameter space two bistability domains. The two stable states are in one case the doughnut modes and in the other the modes TEM(10) and TEM(01). Focusing on the latter case, we study numerically the conditions for optical switching from one mode to the other on injection of an appropriate control beam, and we demonstrate the possibility of using the laser as a NOR logic gate.


Deaths from chronic renal failure under the age of 50. Medical Services Study Group of the Royal College of Physicians.
From a survey of the West Midlands and Mersey Regions and the Grampian Health Board, we found that in 1978 and 1979 some 122 patients with chronic renal failure died in hospital under the age of 50. Of these, 69 had been given dialysis or transplantation, or both, while for many reasons the remainder had been considered unsuitable. While the criteria varied, the reasons given for non-acceptance of cases seemed sound, and in no instance during this particular period was a patient denied dialysis because of a shortage of machines. We think that the public should be aware of these findings and not led to think that if only enough dialysis machines were available death from renal failure would be a rarity.


gamma-Secretase dependent production of intracellular domains is reduced in adult compared to embryonic rat brain membranes.
gamma-Secretase is an intramembrane aspartyl protease whose cleavage of the amyloid precursor protein (APP) generates the amyloid beta-peptide (Abeta) and the APP intracellular domain. Abeta is widely believed to have a causative role in Alzheimer's disease pathogenesis, and therefore modulation of gamma-secretase activity has become a therapeutic goal. Besides APP, more than 50 substrates of gamma-secretase with different cellular functions during embryogenesis as well as adulthood have been revealed. Prior to gamma-secretase cleavage, substrates are ectodomain shedded, producing membrane bound C-terminal fragments (CTFs). Here, we investigated gamma-secretase cleavage of five substrates; APP, Notch1, N-cadherin, ephrinB and p75 neurotrophin receptor (p75-NTR) in membranes isolated from embryonic, young or old adult rat brain by analyzing the release of the corresponding intracellular domains (ICDs) or Abeta40 by western blot analysis and ELISA respectively. The highest levels of all ICDs and Abeta were produced by embryonic membranes. In adult rat brain only cleavage of APP and Notch1 could be detected and the Abeta40 and ICD production from these substrates was similar in young and old adult rat brain. The CTF levels of Notch1, N-cadherin, ephrinB and p75-NTR were also clearly decreased in the adult brain compared to embryonic brain, whereas the APP CTF levels were only slightly decreased. In summary our data suggests that gamma-secretase dependent ICD production is down-regulated in the adult brain compared to embryonic brain. In addition, the present approach may be useful for evaluating the specificity of gamma-secretase inhibitors.


Myd88-dependent positioning of Ptgs2-expressing stromal cells maintains colonic epithelial proliferation during injury.
We identified cellular and molecular mechanisms within the stem cell niche that control the activity of colonic epithelial progenitors (ColEPs) during injury. Here, we show that while WT mice maintained ColEP proliferation in the rectum following injury with dextran sodium sulfate, similarly treated Myd88(-/-) (TLR signaling-deficient) and prostaglandin-endoperoxide synthase 2(-/-) (Ptgs2(-/-)) mice exhibited a profound inhibition of epithelial proliferation and cellular organization within rectal crypts. Exogenous addition of 16,16-dimethyl PGE(2) (dmPGE(2)) rescued the effects of this injury in both knockout mouse strains, indicating that Myd88 signaling is upstream of Ptgs2 and PGE(2). In WT and Myd88(-/-) mice, Ptgs2 was expressed in scattered mesenchymal cells. Surprisingly, Ptgs2 expression was not regulated by injury. Rather, in WT mice, the combination of injury and Myd88 signaling led to the repositioning of a subset of the Ptgs2-expressing stromal cells from the mesenchyme surrounding the middle and upper crypts to an area surrounding the crypt base adjacent to ColEPs. These findings demonstrate that Myd88 and prostaglandin signaling pathways interact to preserve epithelial proliferation during injury using what we believe to be a previously undescribed mechanism requiring proper cellular mobilization within the crypt niche.


Complex social contagion makes networks more vulnerable to disease outbreaks.
Social network analysis is now widely used to investigate the dynamics of infectious disease spread. Vaccination dramatically disrupts disease transmission on a contact network, and indeed, high vaccination rates can potentially halt disease transmission altogether. Here, we build on mounting evidence that health behaviors - such as vaccination, and refusal thereof - can spread across social networks through a process of complex contagion that requires social reinforcement. Using network simulations that model health behavior and infectious disease spread, we find that under otherwise identical conditions, the process by which the health behavior spreads has a very strong effect on disease outbreak dynamics. This dynamic variability results from differences in the topology within susceptible communities that arise during the health behavior spreading process, which in turn depends on the topology of the overall social network. Our findings point to the importance of health behavior spread in predicting and controlling disease outbreaks.


Alignment: a conceptual shift from adherence.
Developing a treatment plan with a patient is one of the cornerstones of clinical medicine. The current concept of adherence, a conceptual shift away from the notion of compliance, connotes an agreement to and implementation of a plan. There are, however, myriad causes that can impinge on whether or not a treatment plan is followed through. The current clinical usage of the word 'adherence' tends to focus on the ultimate behaviour of the patient, and as such the factors acting on the behaviour may escape attention. We propose the concept of treatment 'alignment'. The term 'alignment' conveys an image of the patient and clinician in the context of a range of factors that must 'line up' to maximise the chances of a treatment plan being successfully implemented.


Patients with generalized social phobia direct their attention away from faces.
The experiment tested whether patients with social phobia direct their attention to or away from faces with a range of emotional expressions. A modified dot probe paradigm (J. Abnorm. Psychol. 95 (1986) 15) measured whether participants attended more to faces or to household objects. Twenty patients with social phobia were faster in identifying the probe when it occurred in the location of the household objects, regardless of whether the facial expressions were positive, neutral, or negative. In contrast, controls did not exhibit an attentional preference. The results are in line with recent theories of social phobia that emphasize the role of reduced processing of external social cues in maintaining social anxiety.


Endospore abundance, microbial growth and necromass turnover in deep sub-seafloor sediment.
Two decades of scientific ocean drilling have demonstrated widespread microbial life in deep sub-seafloor sediment, and surprisingly high microbial-cell numbers. Despite the ubiquity of life in the deep biosphere, the large community sizes and the low energy fluxes in this vast buried ecosystem are not yet understood. It is not known whether organisms of the deep biosphere are specifically adapted to extremely low energy fluxes or whether most of the observed cells are in a dormant, spore-like state. Here we apply a new approach--the D:L-amino-acid model--to quantify the distributions and turnover times of living microbial biomass, endospores and microbial necromass, as well as to determine their role in the sub-seafloor carbon budget. The approach combines sensitive analyses of unique bacterial markers (muramic acid and D-amino acids) and the bacterial endospore marker, dipicolinic acid, with racemization dynamics of stereo-isomeric amino acids. Endospores are as abundant as vegetative cells and microbial activity is extremely low, leading to microbial biomass turnover times of hundreds to thousands of years. We infer from model calculations that biomass production is sustained by organic carbon deposited from the surface photosynthetic world millions of years ago and that microbial necromass is recycled over timescales of hundreds of thousands of years.


Upper-mantle water stratification inferred from observations of the 2012 Indian Ocean earthquake.
Water, the most abundant volatile in Earth's interior, preserves the young surface of our planet by catalysing mantle convection, lubricating plate tectonics and feeding arc volcanism. Since planetary accretion, water has been exchanged between the hydrosphere and the geosphere, but its depth distribution in the mantle remains elusive. Water drastically reduces the strength of olivine and this effect can be exploited to estimate the water content of olivine from the mechanical response of the asthenosphere to stress perturbations such as the ones following large earthquakes. Here, we exploit the sensitivity to water of the strength of olivine, the weakest and most abundant mineral in the upper mantle, and observations of the exceptionally large (moment magnitude 8.6) 2012 Indian Ocean earthquake to constrain the stratification of water content in the upper mantle. Taking into account a wide range of temperature conditions and the transient creep of olivine, we explain the transient deformation in the aftermath of the earthquake that was recorded by continuous geodetic stations along Sumatra as the result of water- and stress-activated creep of olivine. This implies a minimum water content of about 0.01 per cent by weight-or 1,600 H atoms per million Si atoms-in the asthenosphere (the part of the upper mantle below the lithosphere). The earthquake ruptured conjugate faults down to great depths, compatible with dry olivine in the oceanic lithosphere. We attribute the steep rheological contrast to dehydration across the lithosphere-asthenosphere boundary, presumably by buoyant melt migration to form the oceanic crust.


Effect of training and familiarity on responsiveness to human cues in domestic dogs (Canis familiaris).
Domestic dogs (Canis familiaris) seem to possess an evolved competency to follow human-given cues, often out-performing their wild progenitor the wolf (Canis lupus) on cue-following tasks. However, domestication may not be solely responsible for the socio-cognitive skills of dogs, with ontogenetic experience also playing a role. This research evaluated the effects of intensive training on cue-following behaviour using an unreinforced object-choice paradigm. The responses of dogs that were trained to competitive levels were compared to those of pet dogs with only basic training, and dogs living in an animal shelter that demonstrated no or only rudimentary following of basic commands. Using a cue-following task where three types of cues were presented by familiar and unfamiliar human partners, the number of cues followed by each training group were recorded. All dogs found cues where gesture was combined with a congruent head and eye movement easier to follow than either gesture or eye gaze alone. Whether the cue-giver was familiar or not had a significant effect on number of cues followed in homed dogs, and the performance of shelter dogs was comparable to the other groups when faced with an unfamiliar cue-giver. Contrary to predictions, level of training did not improve performance on the cue-following task. This work does provide support for the presence of an evolved adaptation to exploit social cues provided by humans that can be augmented by familiarity with the cue giver. However, additional joint activity as experienced in an intensive training regime does not seem to increase accuracy in following human-given cues.


Stress fiber-based movement of Shigella flexneri within cells.
icsA (virG), a gene located on pWR100, the virulence plasmid of Shigella flexneri serotype 5 (M90T), encodes a 120-kDa outer membrane protein. This protein promotes a random intracellular movement of the bacteria and leads to the infection of adjacent cells by the formation of protrusions. This movement, which involves the nucleation, polymerization, and subsequent polarization of actin, is referred to as the Ics phenotype (intra/intercellular spread). Here we present evidence that a second, distinct form of locomotion is also elaborated by S. flexneri in chicken embryo fibroblasts in which the Ics phenotype is not expressed. Using a combination of phase-contrast microcinematography and confocal microscopy, we have demonstrated that bacteria adopt parallel orientations by interacting with stress fibers. This interaction subsequently results in bacterial movement along the stress fibers themselves. This phenomenon occurs independently of the presence of a phagocytic vacuole which is lysed shortly after entry of the bacteria into the cell. It is expressed by M90T and SC560, its icsA mutant. This movement has been termed organelle-like movement (Olm phenotype) and is thought to account for the early accumulation of bacteria seen near the nucleus.


MF63 [2-(6-chloro-1H-phenanthro[9,10-d]imidazol-2-yl)-isophthalonitrile], a selective microsomal prostaglandin E synthase-1 inhibitor, relieves pyresis and pain in preclinical models of inflammation.
Microsomal prostaglandin E synthase-1 (mPGES-1) is a terminal prostaglandin E(2) (PGE(2)) synthase in the cyclooxygenase pathway. Inhibitors of mPGES-1 may block PGE(2) production and relieve inflammatory symptoms. To test the hypothesis, we evaluated the antipyretic and analgesic properties of a novel and selective mPGES-1 inhibitor, MF63 [2-(6-chloro-1H-phenanthro-[9,10-d]imidazol-2-yl)isophthalonitrile], in animal models of inflammation. MF63 potently inhibited the human mPGES-1 enzyme (IC(50) = 1.3 nM), with a high degree (>1000-fold) of selectivity over other prostanoid synthases. In rodent species, MF63 strongly inhibited guinea pig mPGES-1 (IC(50) = 0.9 nM) but not the mouse or rat enzyme. When tested in the guinea pig and a knock-in (KI) mouse expressing human mPGES-1, the compound selectively suppressed the synthesis of PGE(2), but not other prostaglandins inhibitable by nonsteroidal anti-inflammatory drugs (NSAIDs), yet retained NSAID-like efficacy at inhibiting lipopolysaccharide-induced pyresis, hyperalgesia, and iodoacetate-induced osteoarthritic pain. In addition, MF63 did not cause NSAID-like gastrointestinal toxic effects, such as mucosal erosions or leakage in the KI mice or nonhuman primates, although it markedly inhibited PGE(2) synthesis in the KI mouse stomach. Our data demonstrate that mPGES-1 inhibition leads to effective relief of both pyresis and inflammatory pain in preclinical models of inflammation and may be a useful approach for treating inflammatory diseases.


A dark yellow fluorescent protein (YFP)-based Resonance Energy-Accepting Chromoprotein (REACh) for Förster resonance energy transfer with GFP.
Förster resonance energy transfer (FRET) microscopy is a powerful technique that enables the visualization of signaling intermediates, protein interactions, and protein conformational and biochemical status. With the availability of an ever-increasing collection of fluorescent proteins, pairs of spectrally different variants have been used for the study of FRET in living cells. However, suitable spectral overlap, necessary for efficient FRET, is limited by the requirement for proper emission separation. Currently used FRET pairs represent compromises between these opposing spectral demands that reduce the maximally attainable FRET sensitivity. We present a previously undescribed FRET acceptor, a nonfluorescent yellow fluorescent protein (YFP) mutant called REACh (for Resonance Energy-Accepting Chromoprotein). REACh allows the use of the photophysically superior FRET donor EGFP, with which it exhibits optimal spectral overlap, which obviates the need for narrow spectral filtering and allows additional fluorescent labels to be used within the same cell. The latter allows the generation of sophisticated bioassays for complex biological questions. We show that this dark acceptor is ideally suited for donor fluorescence lifetime imaging microscopy (FLIM) and confirm these measurements with an independent intensity-based donor fluorescence quenching resonance energy transfer (FqRET) assay. REACh also can be used in donor photobleaching kinetics-based FRET studies. By detecting FRET between a GFP-tagged ubiquitination substrate and REACh-labeled ubiquitin, we imaged the active ubiquitination machinery inside cells. This assay therefore can be used to study proteins whose function is regulated by ubiquitination.


HMGB1 mediates endogenous TLR2 activation and brain tumor regression.
Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor that carries a 5-y survival rate of 5%. Attempts at eliciting a clinically relevant anti-GBM immune response in brain tumor patients have met with limited success, which is due to brain immune privilege, tumor immune evasion, and a paucity of dendritic cells (DCs) within the central nervous system. Herein we uncovered a novel pathway for the activation of an effective anti-GBM immune response mediated by high-mobility-group box 1 (HMGB1), an alarmin protein released from dying tumor cells, which acts as an endogenous ligand for Toll-like receptor 2 (TLR2) signaling on bone marrow-derived GBM-infiltrating DCs. Using a combined immunotherapy/conditional cytotoxic approach that utilizes adenoviral vectors (Ad) expressing Fms-like tyrosine kinase 3 ligand (Flt3L) and thymidine kinase (TK) delivered into the tumor mass, we demonstrated that CD4(+) and CD8(+) T cells were required for tumor regression and immunological memory. Increased numbers of bone marrow-derived, tumor-infiltrating myeloid DCs (mDCs) were observed in response to the therapy. Infiltration of mDCs into the GBM, clonal expansion of antitumor T cells, and induction of an effective anti-GBM immune response were TLR2 dependent. We then proceeded to identify the endogenous ligand responsible for TLR2 signaling on tumor-infiltrating mDCs. We demonstrated that HMGB1 was released from dying tumor cells, in response to Ad-TK (+ gancyclovir [GCV]) treatment. Increased levels of HMGB1 were also detected in the serum of tumor-bearing Ad-Flt3L/Ad-TK (+GCV)-treated mice. Specific activation of TLR2 signaling was induced by supernatants from Ad-TK (+GCV)-treated GBM cells; this activation was blocked by glycyrrhizin (a specific HMGB1 inhibitor) or with antibodies to HMGB1. HMGB1 was also released from melanoma, small cell lung carcinoma, and glioma cells treated with radiation or temozolomide. Administration of either glycyrrhizin or anti-HMGB1 immunoglobulins to tumor-bearing Ad-Flt3L and Ad-TK treated mice, abolished therapeutic efficacy, highlighting the critical role played by HMGB1-mediated TLR2 signaling to elicit tumor regression. Therapeutic efficacy of Ad-Flt3L and Ad-TK (+GCV) treatment was demonstrated in a second glioma model and in an intracranial melanoma model with concomitant increases in the levels of circulating HMGB1. Our data provide evidence for the molecular and cellular mechanisms that support the rationale for the clinical implementation of antibrain cancer immunotherapies in combination with tumor killing approaches in order to elicit effective antitumor immune responses, and thus, will impact clinical neuro-oncology practice.


[Current status of costs and utilizations of hospital based home health nursing care in Korea].
The purpose of this study was to describe the current status of utilization and costs of home health nursing care by the levels of medical institutes in Korea. A secondary analysis of existing data was used from the national electronic data information(EDI) of 148 home health agencies for 6 months from May to Oct 2005 in total. The 148 agencies had multiple services in cerebral infaction, essential hypertension, sequela of cerebrovascular disease, type 2 diabetes mellitus, etc.. The highest 10 rankings of 76 categories of home health nursing services were composed of 96.4% of the total services, such as simple treatment, inflammatory treatment, urethra & bladder irrigation, inserting indwelling catheter etc., in that order. The highest 20 rankings of 226 categories of home examination services were composed of 77.0% of the total home examination services. In addition, the average cost of home health care per visit was 46,088 Won ( approximately 48 $, 1 $=960 Won). The costs ranged from 74,523 Won ( approximately 78 $, loss of chronic kidney function, N18) to 32,270 Won ( approximately 34 $, other cerebrovascular diseases, I67). Results suggest that client characteristics of hospital based HHNC are not different from community based HHNC or visiting nursing services for elderly. The national results will contribute to baseline data used to establish a policy for the home health nursing care system and education.


Nutrimetabolomics reveals food-specific compounds in urine of adults consuming a DASH-style diet.
Although health benefits of the Dietary Approaches to Stop Hypertension (DASH) diet are established, it is not understood which food compounds result in these benefits. We used metabolomics to identify unique compounds from individual foods of a DASH-style diet and determined if these Food-Specific Compounds (FSC) are detectable in urine from participants in a DASH-style dietary study. We also examined relationships between urinary compounds and blood pressure (BP). Nineteen subjects were randomized into 6-week controlled DASH-style diet interventions. Mass spectrometry-based metabolomics was performed on 24-hour urine samples collected before and after each intervention and on 12 representative DASH-style foods. Between 66-969 compounds were catalogued as FSC; for example, 4-hydroxydiphenylamine was found to be unique to apple. Overall, 13-190 of these FSC were detected in urine, demonstrating that these unmetabolized food compounds can be discovered in urine using metabolomics. Although linear mixed effects models showed no FSC from the 12 profiled foods were significantly associated with BP, other endogenous and food-related compounds were associated with BP (N = 16) and changes in BP over time (N = 6). Overall, this proof of principle study demonstrates that metabolomics can be used to catalog FSC, which can be detected in participant urine following a dietary intervention.


Effect of losartan on performance and physiological responses to exercise at high altitude (5035 m).
Altitude-related and exercise-related elevations in blood pressure (BP) increase the likelihood of developing pulmonary hypertension and high-altitude illness during high-altitude sojourn. This study examined the antihypertensive effect and potential exercise benefit of the angiotensin II receptor antagonist losartan when taken at altitude. Twenty participants, paired for age and ACE genotype status, completed a double-blinded, randomised study, where participants took either losartan (100 mg/day) or placebo for 21 days prior to arrival at 5035 m (Whymper Hut, Mt Chimborazo, Ecuador). Participants completed a maximal exercise test on a supine cycle ergometer at sea level (4 weeks prior) and within 48 hours of arrival to 5035 m (10-day ascent). Power output, beat-to-beat BP, oxygen saturation (SpO2) and heart rate (HR) were recorded during exercise, with resting BP collected from daily medicals during ascent. Before and immediately following exercise at 5035 m, extravascular lung water prevalence was assessed with ultrasound (quantified via B-line count). At altitude, peak power was reduced relative to sea level (p<0.01) in both groups (losartan vs placebo: down 100±29 vs 91±28 W, p=0.55), while SpO2 (70±6 vs 70±5%, p=0.96) and HR (146±21 vs 149±24 bpm, p=0.78) were similar between groups at peak power, as was the increase in systolic BP from rest to peak power (up 80±37 vs 69±33 mm Hg, p=0.56). Exercise increased B-line count (p<0.05), but not differently between groups (up 5±5 vs 8±10, p=0.44). Losartan had no observable effect on resting or exercising BP, exercise-induced symptomology of pulmonary hypertension or performance at 5035 m.


Comorbid diabetes and the risk of progressive chronic kidney disease in HIV-infected adults: data from the Veterans Aging Cohort Study.
Approximately, 15% of HIV-infected individuals have comorbid diabetes. Studies suggest that HIV and diabetes have an additive effect on chronic kidney disease (CKD) progression; however, this observation may be confounded by differences in traditional CKD risk factors. We studied a national cohort of HIV-infected and matched HIV-uninfected individuals who received care through the Veterans Healthcare Administration. Subjects were divided into 4 groups based on baseline HIV and diabetes status, and the rate of progression to an estimated glomerular filtration rate (eGFR) <45 mL/min/1.73m was compared using Cox-proportional hazards modeling to adjust for CKD risk factors. About 31,072 veterans with baseline eGFR ≥45 mL/min/1.73m (10,626 with HIV only, 5088 with diabetes only, and 1796 with both) were followed for a median of 5 years. Mean baseline eGFR was 94 mL/min/1.73m, and 7% progressed to an eGFR < 45 mL/min/1.73m. Compared with those without HIV or diabetes, the relative rate of progression was increased in individuals with diabetes only [adjusted hazard ratio (HR) 2.48; 95% confidence interval (CI): 2.19 to 2.80], HIV only [HR: 2.80, 95% CI: 2.50 to 3.15], and both HIV and diabetes [HR: 4.47, 95% CI: 3.87 to 5.17]. Compared with patients with only HIV or diabetes, patients with both diagnoses are at significantly increased risk of progressive CKD even after adjusting for traditional CKD risk factors. Future studies should evaluate the relative contribution of complex comorbidities and accompanying polypharmacy to the risk of CKD in HIV-infected individuals and prospectively investigate the use of cART, glycemic control, and adjunctive therapy to delay CKD progression.


Effectiveness of restarting lithium treatment after its discontinuation in bipolar I and bipolar II disorders.
This study tested the hypothesis that resumption of lithium treatment of bipolar disorders may be less effective after maintenance treatment has been discontinued. Eighty-six patients with type I or II bipolar disorder, not selected according to response to treatment, were followed prospectively during two periods of lithium maintenance treatment averaging 4.6 and 4.4 years. Morbidity (illness episodes per year, hospitalizations per year, percentage of time ill) was assessed, and use of adjunctive medication was rated. Morbidity was similar in the first and second treatment periods (mean number of episodes = 0.83 and 0.94 per year, respectively; mean percentage of time ill = 18.0% and 24.2%), with no differences in numbers of manic and depressive episodes or differences by gender, diagnostic type, length of first treatment, interval between treatments, or discontinuation rate. There was 12.8% more use of adjunctive medication in the second period. The efficacy of lithium did not differ significantly between the first and second treatment periods.


Anisotropic hydrogen diffusion in α-Zr and Zircaloy predicted by accelerated kinetic Monte Carlo simulations.
This report presents an accelerated kinetic Monte Carlo (KMC) method to compute the diffusivity of hydrogen in hcp metals and alloys, considering both thermally activated hopping and quantum tunneling. The acceleration is achieved by replacing regular KMC jumps in trapping energy basins formed by neighboring tetrahedral interstitial sites, with analytical solutions for basin exiting time and probability. Parameterized by density functional theory (DFT) calculations, the accelerated KMC method is shown to be capable of efficiently calculating hydrogen diffusivity in α-Zr and Zircaloy, without altering the kinetics of long-range diffusion. Above room temperature, hydrogen diffusion in α-Zr and Zircaloy is dominated by thermal hopping, with negligible contribution from quantum tunneling. The diffusivity predicted by this DFT + KMC approach agrees well with that from previous independent experiments and theories, without using any data fitting. The diffusivity along <c> is found to be slightly higher than that along <a>, with the anisotropy saturated at about 1.20 at high temperatures, resolving contradictory results in previous experiments. Demonstrated using hydrogen diffusion in α-Zr, the same method can be extended for on-lattice diffusion in hcp metals, or systems with similar trapping basins.


A reaction-diffusion mechanism influences cell lineage progression as a basis for formation, regeneration, and stability of intestinal crypts.
Colon crypts, a single sheet of epithelia cells, consist of a periodic pattern of stem cells, transit-amplifying cells, and terminally differentiated cells that constantly renew and turnover. Experimental evidence suggests that Wnt signaling promotes and regulates stem cell division, differentiation, and possible cell migrations while intestinal BMP signaling inhibits stem cell self-renewal and repression in crypt formation. As more molecular details on Wnt and BMP in crypts are being discovered, little is still known about how complex interactions among Wnt, BMP, and different types of cells, and surrounding environments may lead to de novo formation of multiple crypts or how such interactions affect regeneration and stability of crypts. We present a mathematical model that contains Wnt and BMP, a cell lineage, and their feedback regulations to study formation, regeneration, and stability of multiple crypts. The computational explorations and linear stability analysis of the model suggest a reaction-diffusion mechanism, which exhibits a short-range activation of Wnt plus a long-range inhibition with modulation of BMP signals in a growing tissue of cell lineage, can account for spontaneous formation of multiple crypts with the spatial and temporal pattern observed in experiments. Through this mechanism, the model can recapitulate some distinctive and important experimental findings such as crypt regeneration and crypt multiplication. BMP is important in maintaining stability of crypts and loss of BMP usually leads to crypt multiplication with a fingering pattern. The study provides a mechanism for de novo formation of multiple intestinal crypts and demonstrates a synergetic role of Wnt and BMP in regeneration and stability of intestinal crypts. The proposed model presents a robust framework for studying spatial and temporal dynamics of cell lineages in growing tissues driven by multiple signaling molecules.


Feasibility study using surface-enhanced Raman spectroscopy for the quantitative detection of excitatory amino acids.
The release of excitatory amino acids (EAAs) from injured neurons has been associated with secondary injury following head trauma. The development of a rapid and sensitive method for the quantification of EAAs may provide a means for clinical management of patients affected by head trauma. We explore the potential application of surface-enhanced Raman spectroscopy (SERS) for rapid quantification of the concentration of EAAs in aqueous silver colloids. The EAAs glutamate (Glu) and aspartate (Asp) are released following head injury and have been observed to exhibit SERS spectra that should enable them to be distinguished in a complex aqueous media. Of the two EAAs, the concentration of Glu has been shown to be more indicative of injury to the central nervous system. Using 30-s scans and a 50-mW argon laser, aqueous Glu is quantifiable from 0.4 to 5 micromol/L and is spectrally distinguishable from Asp. In addition, initial in vivo microdialysis experiments suggest that this SERS system is capable of measuring chemical changes following head trauma in the rat brain. Compared with current high-performance liquid chromatography (HPLC) techniques for amino acid detection, the short scanning and processing time associated with the SERS approach enables measurement on a near-real-time basis, providing clinical information in anticipation of pharmaceutical intervention.


Modelling disease introduction as biological control of invasive predators to preserve endangered prey.
Invasive species are a significant cause of bio-diversity loss particularly in island ecosystems. It has been suggested to release pathogenic parasites as an efficient control measure of these mostly immune-naïve populations. In order to explore the potential impacts of such bio-control approach, we construct and investigate mathematical models describing disease dynamics in a host population that acts as a predator embedded in a simple food chain. The consequences of Feline Immunodeficiency Virus (FIV) introduction into a closed ecosystem are addressed using a bi-trophic system, comprising an indigenous prey (birds) and an introduced predator (cats). Our results show that FIV is unlikely to fully eradicate cats on sub-Antarctic islands, but it can be efficient in depressing their population size, allowing for the recovery of the endangered prey. Depending on the ecological setting and disease transmission mode (we consider proportionate mixing as well as mass action), successful pathogen invasion can induce population oscillations that are not possible in the disease-free predator-prey system. These fluctuations can be seen as a mixed blessing from a management point of view. On the one hand, they may increase the extinction risk of the birds. On the other hand, they provide an opportunity to eradicate cats more easily in combination with other methods such as trapping or culling.


Familial cancer syndromes: a survey.
Several pre-malignant diseases are known to have a genetic etiology. This study focuses attention upon precancerous disorders wherein the mode of inheritance is either well established or wherein it remains unclear even though familial aggregation of the particular diseases have been amply documented. These conditions will be discussed as useful models for systematic investigations of the host etiologic component in carcinogenesis. Our survey of hereditary precancerous syndromes includes multiple polyposis of the coli, the multiple mucosal neuroma syndrome, the Cancer Family Syndrome, Sipple's syndrome, Von Recklinghausen's neurofibromatosus, the multiple nevoid basal cell carcinoma syndrome, tuberous sclerosis, familial cutaneous malignant melanoma, and carcinoma of the breast. We have emphasized the heterogeneous character of many forms of familial cancer. Familial breast cancer associations clearly show such heterogeneity, as do colon cancer syndromes. Certain of these precancerous states are characterized by phenotypes which are clinically apparent, polyposis coli being the classic example. Others, such as Sipple's syndrome are amenable to routine screening for biochemical markers. The bulk of putative genetic cancer-predisposing problems require further basic investigation of modes of inheritance. Cancer control may be enhanced through communication of useful genetic and diagnostic information to primary care physicians. Referral of cancer clusters of possible genetic etiology from clinicians to human geneticists facilitates the necessary basic research.


Probing the faintest stars in a globular star cluster.
NGC 6397 is the second closest globular star cluster to the Sun. Using 5 days of time on the Hubble Space Telescope, we have constructed an ultradeep color-magnitude diagram for this cluster. We see a clear truncation in each of its two major stellar sequences. Faint red main-sequence stars run out well above our observational limit and near to the theoretical prediction for the lowest mass stars capable of stable hydrogen burning in their cores. We also see a truncation in the number counts of faint blue stars, namely white dwarfs. This reflects the limit to which the bulk of the white dwarfs can cool over the lifetime of the cluster. There is also a turn toward bluer colors in the least luminous of these objects. This was predicted for the very coolest white dwarfs with hydrogen-rich atmospheres as the formation of H(2) and the resultant collision-induced absorption cause their atmospheres to become largely opaque to infrared radiation.


α-Difluoromethylornithine reduces gastric carcinogenesis by causing mutations in Helicobacter pylori cagY.
Infection by Helicobacter pylori is the primary cause of gastric adenocarcinoma. The most potent H. pylori virulence factor is cytotoxin-associated gene A (CagA), which is translocated by a type 4 secretion system (T4SS) into gastric epithelial cells and activates oncogenic signaling pathways. The gene cagY encodes for a key component of the T4SS and can undergo gene rearrangements. We have shown that the cancer chemopreventive agent α-difluoromethylornithine (DFMO), known to inhibit the enzyme ornithine decarboxylase, reduces H. pylori-mediated gastric cancer incidence in Mongolian gerbils. In the present study, we questioned whether DFMO might directly affect H. pylori pathogenicity. We show that H. pylori output strains isolated from gerbils treated with DFMO exhibit reduced ability to translocate CagA in gastric epithelial cells. Further, we frequently detected genomic modifications in the middle repeat region of the cagY gene of output strains from DFMO-treated animals, which were associated with alterations in the CagY protein. Gerbils did not develop carcinoma when infected with a DFMO output strain containing rearranged cagY or the parental strain in which the wild-type cagY was replaced by cagY with DFMO-induced rearrangements. Lastly, we demonstrate that in vitro treatment of H. pylori by DFMO induces oxidative DNA damage, expression of the DNA repair enzyme MutS2, and mutations in cagY, demonstrating that DFMO directly affects genomic stability. Deletion of mutS2 abrogated the ability of DFMO to induce cagY rearrangements directly. In conclusion, DFMO-induced oxidative stress in H. pylori leads to genomic alterations and attenuates virulence.


12 GHz passive harmonic mode-locking in a 1.06 μm semiconductor optical amplifier-based fiber laser with figure-eight cavity configuration.
We report the generation of passively harmonic mode-locked pulses using a 1.06 μm semiconductor optical amplifier (SOA) in a figure-eight laser configuration operated in the all-normal-dispersion regime. Different orders of harmonic mode-locking can be obtained from 30 MHz to 12.02 GHz by changing the injection current of the SOA from 80 to 660 mA together with the adjustment of polarization controllers. The highest pulse repetition rate increases almost linearly with the SOA current. As SOA current is set to 660 mA, we obtain the intracavity power of 46 mW at the highest repetition rate of 12.02 GHz, corresponding to the 1202th harmonic of the fundamental mode-locking frequency. To our best knowledge, this is the lowest intracavity power to generate the highest repetition rate with a passively mode-locked laser in the all-normal-dispersion regime.


Diagnostic value of magnetic resonance imaging in perimesencephalic and nonperimesencephalic subarachnoid hemorrhage of unknown origin.
The aim of this study was to evaluate the diagnostic value of MR imaging in perimesencephalic (PM) and nonperimesencephalic (non-PM) subarachnoid hemorrhage (SAH) of unknown origin. The authors conducted a retrospective review of all patients with SAH (1226 patients) in their department between January 1991 and December 2008. Included in the study were cases of spontaneous SAH diagnosed using CT scans obtained within 24 hours of the initial symptoms and initially negative digital subtraction (DS) angiograms. Patients with traumatic SAH and an unknown history were excluded from the study. Patients with initially negative DS angiograms were divided into 2 groups: Group 1, a typically PM bleeding pattern (PM SAH); and Group 2, a non-PM bleeding pattern (non-PM SAH) such as hemorrhage in the sylvian or interhemispheric fissure. Cranial MR imaging including the craniocervical region was performed within 72 hours after SAH was diagnosed in all patients in Groups 1 and 2. One thousand sixty-eight patients underwent DS angiography, and among them were 179 (16.7%) with negative angiograms--47 patients (26.3%) from Group 1 and 132 patients (73.7%) from Group 2. Magnetic resonance imaging demonstrated no bleeding sources in any case (100% negative). Thirty-four patients in Group 1 and 120 patients in Group 2 underwent a second DS angiography study. Digital subtraction angiography revealed an aneurysm as the bleeding source in 1 case in Group 1 and in 13 cases in Group 2. Magnetic resonance imaging of the brain and craniocervical region did not produce additional benefit for the detection of a bleeding source and the therapy administered for PM SAH and non-PM SAH (100% negative). The costs of this examination exceeded the clinical value. Despite the results of this study, MR imaging should be discussed on a case-by-case basis because rare bleeding sources are periodically diagnosed in cases of non-PM SAH. A second-look DS angiogram is necessary because aneurysmal hemorrhage occasionally produces PM SAH as well as non-PM SAH. Further prospective studies are needed to verify the authors' results in the future.


Controlled trial of direct physician order entry: effects on physicians' time utilization in ambulatory primary care internal medicine practices.
Direct physician order entry (POE) offers many potential benefits, but evidence suggests that POE requires substantially more time than traditional paper-based ordering methods. The Medical Gopher is a well-accepted system for direct POE that has been in use for more than 15 years. The authors hypothesized that physicians using the Gopher would not spend any more time writing orders than physicians using paper-based methods. A randomized controlled trial of POE using the Medical Gopher system in 11 primary care internal medicine practices. The authors collected detailed time use data using time motion studies of the physicians and surveyed their opinions about the POE system. The authors found that physicians using the Gopher spent 2.2 min more per patient overall, but when duplicative and administrative tasks were taken into account, physicians were found to have spent only 0.43 min more per patient. With experience, the order entry time fell by 3.73 min per patient. The survey revealed that the physicians believed that the system improved their patient care and wanted the Gopher to continue to be available in their practices. Little extra time, if any, was required for physicians to use the POE system. With experience in its use, physicians may even save time while enjoying the many benefits of POE.


Efficacy of step-oblique mammography for confirmation and localization of densities seen on only one standard mammographic view.
Step-oblique mammography is a technique used to determine with confidence whether a mammographic finding visible on multiple images on only one projection (but not elucidated using standard additional mammographic projections such as the roll view) represents a summation artifact or a true mass, and to precisely localize the true mass for further evaluation (if applicable). This paper describes the step-oblique technique and evaluates its efficacy. Between January 1, 1993 and December 31, 1998, 69 consecutive women underwent step-oblique mammography for the evaluation of densities seen on multiple images in only one standard projection. Additional images were obtained at 15 degrees stepped increments in obliquity. If a one-projection-only finding was not seen on step-oblique images, the density was judged to represent a summation artifact, completing the examination. If a density was visualized and could be triangulated concordantly on step-oblique images ranging from the craniocaudal to the 90 degrees lateral projection, then it was judged to represent a real lesion. Such a lesion was further characterized (mass, neodensity, architectural distortion, focal asymmetric density) and was localized precisely in three dimensions, permitting imaging-guided tissue diagnosis, if appropriate. For all study patients, we also recorded BI-RADS (American College of Radiology Breast Imaging and Data Reporting System) assessment categories; pathology results for biopsied lesions; and mammographic follow-up, clinical follow-up, and linkage to regional tumor registry for nonbiopsied lesions for which at least 2 years had elapsed since step-oblique mammography. Step-oblique mammography differentiated 50 real lesions from 19 summation artifacts. All 50 real lesions, although initially visible on only one standard projection, were successfully localized in three dimensions. Subsequent management resulted in the prompt detection and diagnosis of seven breast cancers and 21 benign lesions. None of the remaining findings managed by follow-up rather than biopsy have subsequently been found to be malignant. Step-oblique mammography is an effective means of evaluating the mammographic finding visible on multiple images on only one standard projection.


Minimally invasive surgery for gastric gastrointestinal stromal tumors.
Minimally invasive surgery has been increasingly performed for gastric gastrointestinal stromal tumors (GIST). In this review we discuss and summarize the current evidence on minimally invasive surgery for gastric GISTs. Laparoscopic resection for gastric GIST has been consistently shown to be associated with superior perioperative outcomes with no compromise in oncological outcomes when compared to open resection in numerous retrospective case-control studies. It has also been shown to be safe and feasible for large tumors or tumors located in unfavorable sites. However, to date, there remains a lack of level 1 evidence from prospective randomized control trials in support of laparoscopic resection.


On TCR binding predictors failing to generalize to unseen peptides.
Several recent studies investigate TCR-peptide/-pMHC binding prediction using machine learning or deep learning approaches. Many of these methods achieve impressive results on test sets, which include peptide sequences that are also included in the training set. In this work, we investigate how state-of-the-art deep learning models for TCR-peptide/-pMHC binding prediction generalize to unseen peptides. We create a dataset including positive samples from IEDB, VDJdb, McPAS-TCR, and the MIRA set, as well as negative samples from both randomization and 10X Genomics assays. We name this collection of samples TChard. We propose the hard split, a simple heuristic for training/test split, which ensures that test samples exclusively present peptides that do not belong to the training set. We investigate the effect of different training/test splitting techniques on the models' test performance, as well as the effect of training and testing the models using mismatched negative samples generated randomly, in addition to the negative samples derived from assays. Our results show that modern deep learning methods fail to generalize to unseen peptides. We provide an explanation why this happens and verify our hypothesis on the TChard dataset. We then conclude that robust prediction of TCR recognition is still far for being solved.


Improved method for estimation of multiple parameters in self-mixing interferometry.
There are two categories of applications for self-mixing interference (SMI)-based sensing: (1) estimation of parameters associated with a semiconductor laser (SL) and (2) measurement of the metrological quantities of the external target. To achieve high resolution sensing, each category of applications requires knowledge from the other. This paper proposes an improved method that can simultaneously measure the parameters of an SL and the target movement in arbitrary form. Starting with the existing SMI model, we derive a new matrix equation for the measurement. The measurement matrix is built by employing all the available data samples obtained from an SMI signal. The total least squares estimation approach is used to estimate the parameters. The proposed method is verified by both simulations and experiments.


Cognitive control of intentions for voluntary actions in individuals with a high level of autistic traits.
Impairments in cognitive control generating deviant adaptive cognition have been proposed to account for the strong preference for repetitive behavior in autism. We examined if this preference reflects intentional deficits rather than problems in task execution in the broader autism phenotype using the Autism-Spectrum Quotient (AQ). Participants chose between two tasks differing in their relative strength by indicating first their voluntary task choice and then responding to the subsequently presented stimulus. We observed a stronger repetition bias for the harder task in high AQ participants, with no other differences between the two groups. These findings indicate that the interference between competing tasks significantly contributes to repetitive behavior in autism by modulating the formation of task intentions when choosing tasks voluntarily.


Older Adults' Casino Gambling Behavior and Their Attitudes Toward New Casino Development.
Research on new casinos typically focuses upon their impact on the community, rather than on specific at-risk groups. This research study explores the impact of the opening of a new casino on attitudes of older adult casino patrons, especially those at particular risk of having gambling problems. Results demonstrate that over 80% of older adult casino patrons would not change their attitudes toward gambling or expect to increase their gambling as a result of the opening of a new casino. However, older adults with problem gambling issues are more likely to indicate they would visit a casino more, spend more time at a casino, and gamble more as a result of the opening of a new casino. In addition, older adults with signs of a gambling problem are more likely to say the opening of a new casino would change their opinions of gambling in general or casino gambling.


Repair versus Checkpoint Functions of BRCA1 Are Differentially Regulated by Site of Chromatin Binding.
The product of the Brca1 tumor-suppressor gene is involved in multiple aspects of the cellular DNA damage response (DDR), including activation of cell-cycle arrests and DNA double-stranded break (DSB) repair by homologous recombination. Prior reports demonstrated that BRCA1 recruitment to areas of DNA breakage depended on RAP80 and the RNF8/RNF168 E3 ubiquitin ligases. Here, we extend these findings by showing that RAP80 is only required for the binding of BRCA1 to regions flanking the DSB, whereas BRCA1 binding directly to DNA breaks requires Nijmegen breakage syndrome 1 (NBS1). These differential recruitment mechanisms differentially affect BRCA1 functions: (i) RAP80-dependent recruitment of BRCA1 to chromatin flanking DNA breaks is required for BRCA1 phosphorylation at serine 1387 and 1423 by ATM and, consequently, for the activation of S and G(2) checkpoints; and (ii) BRCA1 interaction with NBS1 upon DSB induction results in an NBS1-dependent recruitment of BRCA1 directly to the DNA break and is required for nonhomologous end-joining repair. Together, these findings illustrate that spatially distinct fractions of BRCA1 exist at the DSB site, which are recruited by different mechanisms and execute different functions in the DDR.


The relationship between various exposure metrics for elongate mineral particles (EMP) in the taconite mining and processing industry.
Different dimensions of elongate mineral particles (EMP) have been proposed as being relevant to respiratory health end-points such as mesothelioma and lung cancer. In this article, a methodology for converting personal EMP exposures measured using the National Institute for Occupational Safety and Health (NIOSH) 7400/7402 methods to exposures based on other size-based definitions has been proposed and illustrated. Area monitoring for EMP in the taconite mines in Minnesota's Mesabi Iron Range was conducted using a Micro Orifice Uniform Deposit Impactor (MOUDI) size-fractionating sampler. EMP on stages of the MOUDI were counted and sized according to each EMP definition using an indirect-transfer transmission electron microscopy (ISO Method 13794). EMP were identified using energy-dispersive x-ray and electron diffraction analysis. Conversion factors between the EMP counts based on different definitions were estimated using (1) a linear regression model across all locations and (2) a location-specific ratio of the count based on each EMP definition to the NIOSH 7400/7402 count. The highest fractions of EMP concentrations were found for EMP that were 1-3 μm in length and 0.2-0.5 μm in width. Therefore, the current standard NIOSH Method 7400, which only counts EMP >5 μm in length and ≥ 3 in aspect ratio, may underestimate amphibole EMP exposures. At the same time, there was a high degree of correlation between the exposures estimated according to the different size-based metrics. Therefore, the various dimensional definitions probably do not result in different dose-response relationships in epidemiological analyses. Given the high degree of correlation between the various metrics, a result consistent with prior research, a more reasonable metric might be the measurement of all EMP irrespective of size. [Supplementary materials are available for this article. Go to the publisher's online edition of Journal of Occupational and Environmental Hygiene for the following free supplemental resource: figures detailing EMP concentration.].


Species-specific oligonucleotide probes for rRNA of Clostridium difficile and related species.
The large copy number of rRNA makes it an appealing target for oligonucleotide probes designed to identify microorganisms. Given that nucleotide sequences in rRNA are known to reflect phylogeny, species-specific rRNA probes should be feasible if the sequences found in closely related species are different. We sequenced portions of the 16S rRNA of three closely related clostridia found in the human colonic microflora: Clostridium bifermentans, C. sordellii, and C. difficile. The rRNAs of these three species showed 97 to 98% sequence similarity. Five oligonucleotide probes complementary to unique segments of the sequences were end labeled with 32P and hybridized on a nylon filter to the immobilized rRNA of each clostridium. Each probe efficiently hybridized only to the rRNA of the species to which it was directed. Complementary probes emitted a signal that exceeded by a factor of 100 to 1,000 the signal of probes that mismatched the target rRNA by 2 to 5 bases. Even a 1-base difference in rRNA sequence allowed a clear distinction between species. A systematic approach can efficiently yield taxon-specific oligonucleotide probes directed at rRNA.


RAMESES II reporting standards for realist evaluations.
Realist evaluation is increasingly used in health services and other fields of research and evaluation. No previous standards exist for reporting realist evaluations. This standard was developed as part of the RAMESES II project. The project's aim is to produce initial reporting standards for realist evaluations. We purposively recruited a maximum variety sample of an international group of experts in realist evaluation to our online Delphi panel. Panel members came from a variety of disciplines, sectors and policy fields. We prepared the briefing materials for our Delphi panel by summarising the most recent literature on realist evaluations to identify how and why rigour had been demonstrated and where gaps in expertise and rigour were evident. We also drew on our collective experience as realist evaluators, in training and supporting realist evaluations, and on the RAMESES email list to help us develop the briefing materials. Through discussion within the project team, we developed a list of issues related to quality that needed to be addressed when carrying out realist evaluations. These were then shared with the panel members and their feedback was sought. Once the panel members had provided their feedback on our briefing materials, we constructed a set of items for potential inclusion in the reporting standards and circulated these online to panel members. Panel members were asked to rank each potential item twice on a 7-point Likert scale, once for relevance and once for validity. They were also encouraged to provide free text comments. We recruited 35 panel members from 27 organisations across six countries from nine different disciplines. Within three rounds our Delphi panel was able to reach consensus on 20 items that should be included in the reporting standards for realist evaluations. The overall response rates for all items for rounds 1, 2 and 3 were 94 %, 76 % and 80 %, respectively. These reporting standards for realist evaluations have been developed by drawing on a range of sources. We hope that these standards will lead to greater consistency and rigour of reporting and make realist evaluation reports more accessible, usable and helpful to different stakeholders.