Protein profiling of microdissected pancreas carcinoma and identification of HSP27 as a potential serum marker.
Patients with pancreatic adenocarcinomas have a poor prognosis because of late clinical manifestation and the tumor's aggressive nature. We used proteomic techniques to search for markers of pancreatic carcinoma. We performed protein profiling of microdissected cryostat sections of 9 pancreatic adenocarcinomas and 10 healthy pancreatic tissue samples using ProteinChip technology (surface-enhanced laser desorption/ionization). We identified proteins by use of 2-dimensional gel electrophoresis, peptide fingerprint mapping, and immunodepletion and used immunohistochemistry for in situ localization of the proteins found. We used ELISA to quantify these proteins in preoperative serum samples from 35 patients with pancreatic cancer and 37 healthy individuals. From among the differentially expressed signals that were detected by ProteinChip technology, we identified 2 proteins, DJ-1 and heat shock protein 27 (HSP27). We then detected HSP27 in sera of patients by use of ELISA, indicating a sensitivity of 100% and a specificity of 84% for the recognition of pancreatic cancer. The detection of DJ-1 and HSP27 in pure defined tissue and the retrieval of HSP27 in serum by antibody-based methods identifies a potential marker for pancreatic cancer.


Regulation of p53 activity through lysine methylation.
p53 is a tumour suppressor that regulates the cellular response to genotoxic stresses. p53 is a short-lived protein and its activity is regulated mostly by stabilization via different post-translational modifications. Here we report a novel mechanism of p53 regulation through lysine methylation by Set9 methyltransferase. Set9 specifically methylates p53 at one residue within the carboxyl-terminus regulatory region. Methylated p53 is restricted to the nucleus and the modification positively affects its stability. Set9 regulates the expression of p53 target genes in a manner dependent on the p53-methylation site. The crystal structure of a ternary complex of Set9 with a p53 peptide and the cofactor product S-adenosyl-l-homocysteine (AdoHcy) provides the molecular basis for recognition of p53 by this lysine methyltransferase.


Coming up short: Comparing venous blood, dried blood spots & saliva samples for measuring telomere length in health equity research.
Telomere length (TL) in peripheral blood mononuclear cells (PBMC) from fresh venous blood is increasingly used to estimate molecular impacts of accumulated social adversity on population health. Sometimes, TL extracted from saliva or dried blood spots (DBS) are substituted as less invasive and more scalable specimen collection methods; yet, are they interchangeable with fresh blood? Studies find TL is correlated across tissues, but have not addressed the critical question for social epidemiological applications: Do different specimen types show the same association between TL and social constructs? We integrate expertise in social epidemiology, molecular biology, and the statistical impact of measurement error on parameter estimates. Recruiting a diverse sample of 132 Metro-Detroit women, we measure TL for each woman from fresh blood PBMC, DBS, and saliva. Using regression methods, we estimate associations between social characteristics and TL, comparing estimates across specimen types for each woman. Associations between TL and social characteristics vary by specimen type collected from the same woman, sometimes qualitatively altering estimates of the magnitude or direction of a theorized relationship. Being Black is associated with shorter TL in PBMC, but longer TL in saliva or DBS. Education is positively associated with TL in fresh blood, but negatively associated with TL using DBS. Findings raise concerns about the use of TL measures derived from different tissues in social epidemiological research. Investigators need to consider the possibility that associations between social variables and TL may be systematically related to specimen type, rather than be valid indicators of socially-patterned biopsychosocial processes.


TMEM16F forms a Ca2+-activated cation channel required for lipid scrambling in platelets during blood coagulation.
Collapse of membrane lipid asymmetry is a hallmark of blood coagulation. TMEM16F of the TMEM16 family that includes TMEM16A/B Ca(2+)-activated Cl(-) channels (CaCCs) is linked to Scott syndrome with deficient Ca(2+)-dependent lipid scrambling. We generated TMEM16F knockout mice that exhibit bleeding defects and protection in an arterial thrombosis model associated with platelet deficiency in Ca(2+)-dependent phosphatidylserine exposure and procoagulant activity and lack a Ca(2+)-activated cation current in the platelet precursor megakaryocytes. Heterologous expression of TMEM16F generates a small-conductance Ca(2+)-activated nonselective cation (SCAN) current with subpicosiemens single-channel conductance rather than a CaCC. TMEM16F-SCAN channels permeate both monovalent and divalent cations, including Ca(2+), and exhibit synergistic gating by Ca(2+) and voltage. We further pinpointed a residue in the putative pore region important for the cation versus anion selectivity of TMEM16F-SCAN and TMEM16A-CaCC channels. This study thus identifies a Ca(2+)-activated channel permeable to Ca(2+) and critical for Ca(2+)-dependent scramblase activity during blood coagulation. PAPERFLICK:


Novel quantitative TaqMan® MGB real-time PCR for sensitive detection of Vibrio aestuarianus in Crassostrea gigas.
Validation of a novel quantitative real-time PCR using TaqMan® minor groove binder (MGB) chemistry is described for sensitive and rapid detection of Vibrio aestuarianus, an increasingly important pathogen of Pacific cupped oyster Crassostrea gigas aquaculture. Primers and TaqMan® MGB hydrolysis probe were designed to specifically amplify a 58bp DNA fragment of the V. aestuarianus dnaJ gene. Real-time PCR selectivity was empirically tested using DNA extracted from isolates of V. aestuarianus and a selection of different aquatic bacterial species, including other Vibrio spp. Theoretical selectivity was assessed through sequence comparison using the NCBI BLAST similarity tool. Quantitative PCR plasmid standards were generated to test assay linearity, amplification efficiency and the limit of quantitation (LOQ), according to International Organisation for Standardisation ISO 16140 validation recommendations. LOQ ranged between 5 and 10 PCR copies, although the detection range extended beyond this with reduced precision. Applied performance was tested using C. gigas samples taken from a selection of Irish aquaculture sites. Increasing levels of V. aestuarianus, accompanied by the development of tissue pathology in examined oysters, were found at 1 site that was sampled repeatedly in 2013. Rapid, sensitive and reproducible detections of V. aestuarianus from C. gigas tissue samples were attained during this validation study with a small sample size, and a practical application for disease management is described.


Ecotoxicity test methods for engineered nanomaterials: practical experiences and recommendations from the bench.
Ecotoxicology research is using many methods for engineered nanomaterials (ENMs), but the collective experience from researchers has not been documented. This paper reports the practical issues for working with ENMs and suggests nano-specific modifications to protocols. The review considers generic practical issues, as well as specific issues for aquatic tests, marine grazers, soil organisms, and bioaccumulation studies. Current procedures for cleaning glassware are adequate, but electrodes are problematic. The maintenance of exposure concentration is challenging, but can be achieved with some ENMs. The need to characterize the media during experiments is identified, but rapid analytical methods are not available to do this. The use of sonication and natural/synthetic dispersants are discussed. Nano-specific biological endpoints may be developed for a tiered monitoring scheme to diagnose ENM exposure or effect. A case study of the algal growth test highlights many small deviations in current regulatory test protocols that are allowed (shaking, lighting, mixing methods), but these should be standardized for ENMs. Invertebrate (Daphnia) tests should account for mechanical toxicity of ENMs. Fish tests should consider semistatic exposure to minimize wastewater and animal husbandry. The inclusion of a benthic test is recommended for the base set of ecotoxicity tests with ENMs. The sensitivity of soil tests needs to be increased for ENMs and shortened for logistics reasons; improvements include using Caenorhabditis elegans, aquatic media, and metabolism endpoints in the plant growth tests. The existing bioaccumulation tests are conceptually flawed and require considerable modification, or a new test, to work for ENMs. Overall, most methodologies need some amendments, and recommendations are made to assist researchers.


Pulmonary Nodule Detection in CT Images: False Positive Reduction Using Multi-View Convolutional Networks.
We propose a novel Computer-Aided Detection (CAD) system for pulmonary nodules using multi-view convolutional networks (ConvNets), for which discriminative features are automatically learnt from the training data. The network is fed with nodule candidates obtained by combining three candidate detectors specifically designed for solid, subsolid, and large nodules. For each candidate, a set of 2-D patches from differently oriented planes is extracted. The proposed architecture comprises multiple streams of 2-D ConvNets, for which the outputs are combined using a dedicated fusion method to get the final classification. Data augmentation and dropout are applied to avoid overfitting. On 888 scans of the publicly available LIDC-IDRI dataset, our method reaches high detection sensitivities of 85.4% and 90.1% at 1 and 4 false positives per scan, respectively. An additional evaluation on independent datasets from the ANODE09 challenge and DLCST is performed. We showed that the proposed multi-view ConvNets is highly suited to be used for false positive reduction of a CAD system.


An exploration and comparison of food and drink availability in homes in a sample of families of White and Pakistani origin within the UK.
Knowledge of the types and quantities of foods and drinks available in family homes supports the development of targeted intervention programmes for obesity prevention or management, or for overall diet improvement. In the UK, contemporary data on foods that are available within family homes are lacking. The present study aimed to explore home food and drink availability in UK homes. An exploratory study using researcher-conducted home food availability inventories, measuring all foods and drinks within the categories of fruits, vegetables, snack foods and beverages. Bradford, a town in the north of the UK. Opportunistic sample of mixed ethnicity families with infants approximately 18 months old from the Born in Bradford birth cohort. All homes had at least one type of fruit, vegetable and snack available. Fresh fruits commonly available were oranges, bananas, apples, satsumas and grapes. Commonly available fresh vegetables included potatoes, cucumber, tomatoes and carrots. The single greatest non-fresh fruit available in homes was raisins. Non-fresh vegetables contributing the most were frozen mixed vegetables, tinned tomatoes and tinned peas. Ethnic differences were found for the availability of fresh fruits and sugar-sweetened beverages, which were both found in higher amounts in Pakistani homes compared with White homes. These data contribute to international data on availability and provide an insight into food availability within family homes in the UK. They have also supported a needs assessment of the development of a culturally specific obesity prevention intervention in which fruits and vegetables and sugar-sweetened beverages are targeted.


Agalsidase therapy in patients with Fabry disease on renal replacement therapy: a nationwide study in Italy.
In Fabry disease, end-stage renal disease (ESRD) and severe neurologic and cardiac complications represent the leading causes of late morbidity and mortality. A comprehensive Italian nationwide survey study was conducted to explore changes in cardiac status and renal allograft function in Fabry patients on renal replacement therapy (RRT) and enzyme replacement therapy (ERT). This study was designed as a cross-sectional survey study with prospective follow-up. Of the 34 patients identified via searches in registries, 31 males and 2 females who received RRT and ERT (agalsidase beta in 30 patients, agalsidase alpha in 3) were included. Left ventricular mass index (LVMI), interventricular septal thickness at end diastole (IVSD), left ventricular posterior wall thickness (LVPWT) and renal allograft function were assessed at ERT baseline and subsequently at yearly intervals. The patients in the dialysis and transplant groups had been started on dialysis at age 42.0 and 37.1 years (mean), respectively, and patients in the transplant group received their renal allograft at age 39.8 years (mean). The mean age at the start of ERT was similar, 44.1 and 44.6 years, respectively. The mean RRT follow-up was 61.1 and 110.6 months for dialysis and transplant patients, respectively, whereas the ERT duration was 45.1 and 48.4 months, respectively. Cardiac parameters increased in dialysis patients. In transplant patients, mean LVMI seemed to plateau during agalsidase therapy at a lower level as compared to baseline. Decline in renal allograft function was relatively mild (-1.92 ml/min/year). Agalsidase therapy was well tolerated. Serious ERT-unrelated events occurred more often in the dialysis group. Kidney transplantation should be the standard of care for Fabry patients progressing towards ESRD. Transplanted Fabry patients on ERT may do better than patients remaining on maintenance dialysis. Larger, controlled studies in Fabry patients with ESRD will have to demonstrate if ERT is able to change the trajectory of cardiac disease and can preserve graft renal function.


Telomere length and pancreatic cancer: a case-control study.
Telomeres, the ends of chromosomes, are critical for maintaining genomic stability and grow shorter with age. Shortened telomeres in pancreatic tissue play a key role in the pathogenesis of pancreatic cancer, and shorter telomeres in peripheral blood leukocytes (PBL) have been associated with increased risk for several cancer types. We hypothesized that shorter blood telomeres are associated with higher risk for pancreatic cancer. Telomere length was measured in PBLs using quantitative real-time PCR in 499 cases with pancreatic cancer and 963 cancer-free controls from the Mayo Clinic. ORs and confidence intervals (CI) were computed using logistic generalized additive models (GAM) adjusting for multiple variables. In multivariable adjusted models, we observed a significant nonlinear association between telomere length in peripheral blood samples and the risk for pancreatic cancer. Risk was lower among those with longer telomeres compared with shorter telomeres across a range from the 1st percentile to 90th percentile of telomere length. There was also some evidence for higher risk among those with telomeres in the longest extreme. Short telomeres in peripheral blood are associated with an increased risk for pancreatic cancer across most of the distribution of length, but extremely long telomeres may also be associated with higher risk. Although the temporality of this relationship is unknown, telomere length may be useful as either a marker of pancreatic cancer risk or of the presence of undetected pancreatic cancer. If telomere shortening precedes cancer incidence, interventions to preserve telomere length may be an effective strategy to prevent pancreatic cancer.


On the relative importance of individual-level characteristics and dyadic interaction effects in negotiations: Variance partitioning evidence from a twins study.
Negotiations are inherently dyadic. Negotiators' individual-level characteristics may not only make them perform better or worse in general, but also may make them particularly well- or poorly-suited to negotiate with a particular counterpart. The present research estimates the extent to which performance in a distributive negotiation is affected by (a) the negotiators' individual-level characteristics and (b) dyadic interaction effects that are defined by the unique pairings between the negotiators and their counterparts. Because negotiators cannot interact multiple times without carryover effects, we estimated the relative importance of these factors with a new methodology that used twin siblings as stand-ins for each other. Participants engaged in a series of 1-on-1 negotiations with counterparts while, elsewhere, their cotwins engaged in the same series of 1-on-1 negotiations with the cotwins of those counterparts. In these data, dyadic interaction effects explained more variation in negotiation economic outcomes than did individual differences, whereas individual differences explain more than twice as much of the variation in subjective negotiation outcomes than did dyadic interaction effects. These results suggest dyadic interaction effects represent an understudied area for future research, particularly with regard to the economic outcomes of negotiations. (PsycINFO Database Record


Exacerbation of reperfusion arrhythmias by sudden oxidant stress.
A burst of free oxygen radical production has been demonstrated during the early moments of reperfusion, coincident with the onset of reperfusion arrhythmias, which can be attenuated by antioxidants. We have investigated whether a sudden burst of oxidant stress, superimposed on that occurring during reperfusion, can exacerbate reperfusion arrhythmias. Rat hearts (n = 12/group) were subjected to 12 minutes of aerobic perfusion; during the last 2 minutes, rose bengal (1 mumol/l) was added to the perfusion fluid. Then, regional ischemia was induced, and rose bengal-free perfusion was restored. After 5 minutes of ischemia, reperfusion was initiated for 5 minutes, and during the first 30 seconds of reperfusion, hearts were uniformly illuminated (8,500 lux) with green light (530-590 nm). The photoactivation of rose bengal trapped in the tissue, producing singlet oxygen and superoxide, resulted in an exacerbation of reperfusion arrhythmias. Thus, 92% of hearts developed ventricular premature beats, 83% ventricular tachycardia, and 33% ventricular fibrillation. In contrast, hearts with regional ischemia and reperfusion in the absence of rose bengal and/or illumination did not develop ventricular fibrillation; only one heart exhibited ventricular tachycardia, and the incidence of ventricular premature beats was lower (42-50%). Furthermore, the burst of oxidant stress shortened the time to onset of ventricular premature beats from 21.7 +/- 5.6 to 9.9 +/- 2.1 seconds. Additional studies revealed that rose bengal photoactivation without reperfusion was less arrhythmogenic compared with the combination of reperfusion plus photoactivation. These results demonstrate that a sudden burst of oxidant stress during the early moments of reperfusion can exacerbate the vulnerability to reperfusion arrhythmias.


Prediction of data stream parameters in atmospheric turbulent wireless communication links.
A unified approach for calculation of information data stream parameters in the atmospheric optical communication channel is presented based on irradiance fluctuations of optical wave propagation through turbulence and on a generalized Ricean K-parameter distribution. The effects of turbulence are described via the well-known Kolmogorov scheme of turbulent structure relaxation in terms of stochastic scintillation theory described by the gamma-gamma distribution along with measurements of the values of the refractive index structure parameter, C(n)(2). The relation between the Ricean parameter K and the signal scintillation parameter sigma(I)(2) is considered to develop a unified description of the corresponding probability density function (pdf) of signal fading within an atmospheric wireless communication link. Through the corresponding pdf and parameter K, signal data stream parameters such as the signal-to-noise ratio (SNR), bit error rate (BER), and capacity of the optical atmospheric channel (C) are estimated. Such an approach permits the reliable prediction of the effects of fading caused by different levels of turbulence and agrees with experimental data observed at different atmospheric levels, at the heights of both 100-200 m and above 1-2 km. It is shown that at heights of 100-200 m, effects of fading, caused by turbulence, occur much more frequently than those at the heights of 1-2 km. Data stream parameters such as channel capacity, SNR, and spectral efficiency become stronger at higher altitudes, while at the same time the BER becomes relatively negligible.


Heterogeneity in children's developing moral judgments about different types of harm.
The patterning of 160 U.S. 4- to 9-year-olds' (M = 6.23 years, SD = 1.46) moral judgments regarding physical harm, psychological harm, and unfair resource distribution transgressions were examined in separate latent profile analyses. Judgments regarding physical harm yielded a single Prototypical profile, where transgressions were judged as very unacceptable, punishable, wrong even if not regulated, intentional, harmful, and as causing very negative emotions for victims. A similar Prototypical profile, as well as an Emergent profile, was extracted for both unfair resource distribution and psychological harm; children in the Emergent profile evaluated transgressions as more acceptable, particularly when rules and authority were removed, less punishable, less intentional, and less harmful than did Prototypical children. In addition, psychological harm was characterized by a small Inconsistent profile, where children judged moral transgressions as very acceptable but also as intentional and harmful. Profiles were differentiated by child age and authority independence justifications, but not by individual differences in social competence, gender, or act acceptability justifications. Children in the Inconsistent profile for psychological harm or in the Emergent profile for unfair resource distribution were younger than children in the other profile(s) for the respective harm type. Children who were classified in the Prototypical (or the Emergent) profile when judging unequal distribution were more likely than would be expected by chance to be classified in the equivalent profile when evaluating psychological harm transgressions. The results show that moral judgments are organized and reflect age differences in children's moral understanding of different types of harms. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB 1954) by human NAD(P)H quinone oxidoreductase 2: a novel co-substrate-mediated antitumor prodrug therapy.
A novel prodrug activation system, endogenous in human tumor cells, is described. A latent enzyme-prodrug system is switched on by a simple synthetic, small molecule co-substrate. This ternary system is inactive if any one of the components is absent. CB 1954 [5-(aziridin-1-yl)-2,4-dinitrobenzamide] is an antitumor prodrug that is activated in certain rat tumors via its 4-hydroxylamine derivative to a potent bifunctional alkylating agent. However, human tumor cells are resistant to CB 1954 because they are unable to catalyze this bioactivation efficiently. A human enzyme has been discovered that can activate CB 1954, and it has been shown to be commonly present in human tumor cells. The enzyme is NQO2 [NAD(P)H quinone oxidoreductase 2], but its activity is normally latent, and a nonbiogenic co-substrate such as NRH [nicotinamide riboside (reduced)] is required for enzymatic activity. There is a very large (100-3000-fold) increase in CB 1954 cytotoxicity toward either NQO2-transfected rodent or nontransfected human tumor cell lines in the presence of NRH. Other reduced pyridinium compounds can also act as co-substrates for NQO2. Thus, the simplest quaternary salt of nicotinamide, 1-methyl-3-carboxamidopyridinium iodide, was a co-substrate for NQO2 when reduced to the corresponding 1,4-dihydropyridine derivative. Increased chain length and/or alkyl load at the 1-position of the dihydropyridine ring improved specific activity, and compounds more active than NRH were found. However, little activity was seen with either the 1-benzyl or 1-(2-phenylethyl) derivatives. A negatively charged substituent at the 3-position of the reduced pyridine ring also negated the ability of these compounds to act as cosubstrates for NQO2. In particular, 1-carbamoylmethyl-3-carbamoyl-1,4dihydropyridine was shown to be a co-substrate for NQO2 with greater stability than NRH, with the ability to enter cells and potentiate the cytotoxicity of CB 1954. Furthermore, this agent is synthetically accessible and suitable for further pharmaceutical development. NQO2 activity appears to be related to expression of NQO1 (DT-diaphorase), an enzyme that is known to have a favorable distribution toward certain human cancers. NQO2 is a novel target for prodrug therapy and has a unique activation mechanism that relies on a synthetic co-substrate to activate an apparently latent enzyme. Our findings may reopen the use of CB 1954 for the direct therapy of human malignant disease.


Expression of drug targets in primary and matched metastatic renal cell carcinoma tumors.
Targeted therapies in renal cell carcinoma can have different effects on primary and metastatic tumors. To pave the way for predictive biomarker development, we assessed differences in expression of targets of currently approved drugs in matched primary and metastatic specimens from 34 patients. Four cores from each site were embedded in tissue microarray blocks. Expression of B-Raf, C-Raf, cKIT, FGF-R1, HIF-2α, mTOR, PDGF-Rβ, VEGF-R1, VEGF-R2, VEGF-R3, VEGF, VEGF-B, VEGF-C, VEGF-D, MEK1, and ERK1/2 was studied using a quantitative immunofluorescence method. No significant differences were observed in global expression levels in primary and metastatic renal cell carcinoma tumors, with the exception of MEK, which had higher expression in metastatic than primary specimens. Similarly, more ki67 positive cells were seen in metastatic specimens. Correlations between marker expression in primary and metastatic specimens were variable, with the lowest correlation seen for FGF-R1 and VEGF-D. There were no significant differences in the degree of heterogeneity in primary versus metastatic tumors. Expression of most of the studied markers was similar in primary and metastatic renal cell carcinoma tumors, suggesting that predictive biomarker testing for these markers can be conducted on either the primary or metastatic tumors for most markers.


Cardiovascular disease in kidney donors: matched cohort study.
To determine whether people who donate a kidney have an increased risk of cardiovascular disease. Retrospective population based matched cohort study. All people who were carefully selected to become a living kidney donor in the province of Ontario, Canada, between 1992 and 2009. The information in donor charts was manually reviewed and linked to provincial healthcare databases. Matched non-donors were selected from the healthiest segment of the general population. A total of 2028 donors and 20,280 matched non-donors were followed for a median of 6.5 years (maximum 17.7 years). Median age was 43 at the time of donation (interquartile range 34-50) and 50 at the time of follow-up (42-58). The primary outcome was a composite of time to death or first major cardiovascular event. The secondary outcome was time to first major cardiovascular event censored for death. The risk of the primary outcome of death and major cardiovascular events was lower in donors than in non-donors (2.8 v 4.1 events per 1000 person years; hazard ratio 0.66, 95% confidence interval 0.48 to 0.90). The risk of major cardiovascular events censored for death was no different in donors than in non-donors (1.7 v 2.0 events per 1000 person years; 0.85, 0.57 to 1.27). Results were similar in all sensitivity analyses. Older age and lower income were associated with a higher risk of death and major cardiovascular events in both donors and non-donors when each group was analysed separately. The risk of major cardiovascular events in donors is no higher in the first decade after kidney donation compared with a similarly healthy segment of the general population. While we will continue to follow people in this study, these interim results add to the evidence base supporting the safety of the practice among carefully selected donors.


Shared care for high-dependency patients: mental illness, neurological disorders and terminal care--a review.
Changes in the structure of health services in the UK have increased the need to identify and formalize shared responsibilities between primary and secondary care for patients with chronic conditions. There are well-established schemes for the management of patients with some chronic diseases but very little for other high-dependency groups. This review examines the extent of systematic and shared care for some of the less well served groups: these are mental illness, neurological disorders and terminal care. Examples of good practice are highlighted.


Identification of a transmembrane glycoprotein specific for secretory vesicles of neural and endocrine cells.
Several types of cells store proteins in secretory vesicles from which they are released by an appropriate stimulus. It might be expected that the secretory vesicles in different cell types use similar molecular machinery. Here we describe a transmembrane glycoprotein (Mr approximately 100,000) that is present in secretory vesicles in all neurons and endocrine cells studied, in species from elasmobranch fish to mammals, and in neural and endocrine cell lines. It was detected by cross-reactivity with monoclonal antibodies raised to highly purified cholinergic synaptic vesicles from the electric organ of fish. By immunoprecipitation of intact synaptic vesicles and electron microscopic immunoperoxidase labeling, we have shown that the antigenic determinant is on the cytoplasmic face of the synaptic vesicles. However, the electrophoretic mobility of the antigen synthesized in the presence of tunicamycin is reduced to Mr approximately 62,000, which suggests that the antigen is glycosylated and must therefore span the vesicle membrane.


Reward-Modulated Response Inhibition, Cognitive Shifting, and the Orbital Frontal Cortex in Early Adolescence.
Immaturities in cognitive shifting are associated with adolescent risk behaviors. The orbital frontal cortex (OFC) regulates reward processing and response inhibition. This study tested the relationship between cognitive shifting, OFC activity, and reward-modulated response inhibition in young adolescents. An fMRI antisaccade (AS) paradigm examined the effects of reward conditions on inhibitory response and OFC processing. A validated self-report inventory assessed cognitive shifting. Compared to neutral, reward trials showed better AS performance and increased OFC activation. Cognitive shifting positively associated with AS performance in reward and neutral trials. Poorer cognitive shifting predicted greater OFC activation. Results indicate lower OFC efficiency, as greater activation to achieve correct performance, underlies cognitive shifting problems. These neurocognitive impairments are relevant for understanding adolescent risk behaviors.


Accounting for individual differences in human associative learning.
Associative learning has provided fundamental insights to understanding psychopathology. However, psychopathology occurs along a continuum and as such, identification of disruptions in processes of associative learning associated with aspects of psychopathology illustrates a general flexibility in human associative learning. A handful of studies have looked specifically at individual differences in human associative learning, but while much work has concentrated on accounting for flexibility in learning caused by external factors, there has been limited work considering how to model the influence of dispositional factors. This review looks at the range of individual differences in human associative learning that have been explored and the attempts to account for, and model, this flexibility. To fully understand human associative learning, further research needs to attend to the causes of variation in human learning.


Expression of adhesion molecules, monocyte interactions and oxidative stress in human endothelial cells exposed to wood smoke and diesel exhaust particulate matter.
Toxicological effects of wood smoke particles are less investigated than traffic-related combustion particles. We investigated the effect of wood smoke particles, generated by smouldering combustion conditions, on human umbilical endothelial cells (HUVECs) co-cultured with or without monocytic THP-1 cells. Standard reference material (SRM) 2975 diesel exhaust particles were used as benchmark particles. Wood smoke particles at 50 μg/ml or 100 μg/ml caused adhesion of THP-1 monocytes onto HUVECs in co-cultures, whereas SRM2975 had no such effect. Both types of particles from 1 μg/ml increased VCAM-1 expression on HUVECs in mono-cultures. However, only the exposure to wood smoke particles was associated with increased expression of TNF and IL8 mRNA in THP-1 cells. We found no effect on the intracellular production of reactive oxygen species by the fluorescent probe DCFH-DA, whereas especially the wood smoke particles caused increased level of DNA strand breaks and oxidised guanines at concentrations with low cytotoxicity. In conclusion, our results indicate that the adherence of monocytes on endothelial cells in wood smoke particle exposed cultures depend on activation of both cell types.


Investigating the Feasibility of Targeted Next-Generation Sequencing to Guide the Treatment of Head and Neck Squamous Cell Carcinoma.
Head and neck squamous cell carcinoma (HNSCC) is a deadly disease in which precision medicine needs to be incorporated. We aimed to implement next-generation sequencing (NGS) in determining actionable targets to guide appropriate molecular targeted therapy in HNSCC patients. Ninety-three tumors and matched blood samples underwent targeted sequencing of 244 genes using the Illumina HiSeq 2500 platform with an average depth of coverage of greater than 1,000×. Clinicopathological data from patients were obtained from 17 centers in Korea, and were analyzed in correlation with NGS data. Ninety-two of the 93 tumors were amenable to data analysis. TP53 was the most common mutation, occurring in 47 (51%) patients, followed by CDKN2A (n=23, 25%), CCND1 (n=22, 24%), and PIK3CA (n=19, 21%). The total mutational burden was similar between human papillomavirus (HPV)-negative vs. positive tumors, although TP53, CDKN2A and CCND1 gene alterations occurred more frequently in HPV-negative tumors. HPV-positive tumors were significantly associated with immune signature-related genes compared to HPV-negative tumors. Mutations of NOTCH1 (p=0.027), CDKN2A (p < 0.001), and TP53 (p=0.038) were significantly associated with poorer overall survival. FAT1 mutations were highly enriched in cisplatin responders, and potentially targetable alterations such as PIK3CA E545K and CDKN2A R58X were noted in 14 patients (15%). We found several targetable genetic alterations, and our findings suggest that implementation of precision medicine in HNSCC is feasible. The predictive value of each targetable alteration should be assessed in a future umbrella trial using matched molecular targeted agents.


Transcriptional biomarkers--high throughput screening, quantitative verification, and bioinformatical validation methods.
Molecular biomarkers found their way into many research fields, especially in molecular medicine, medical diagnostics, disease prognosis, risk assessment but also in other areas like food safety. Different definitions for the term biomarker exist, but on the whole biomarkers are measureable biological molecules that are characteristic for a specific physiological status including drug intervention, normal or pathological processes. There are various examples for molecular biomarkers that are already successfully used in clinical diagnostics, especially as prognostic or diagnostic tool for diseases. Molecular biomarkers can be identified on different molecular levels, namely the genome, the epigenome, the transcriptome, the proteome, the metabolome and the lipidome. With special "omic" technologies, nowadays often high throughput technologies, these molecular biomarkers can be identified and quantitatively measured. This article describes the different molecular levels on which biomarker research is possible including some biomarker candidates that have already been identified. Hereby the transcriptomic approach will be described in detail including available high throughput methods, molecular levels, quantitative verification, and biostatistical requirements for transcriptional biomarker identification and validation.


Differential effect of 5- and 15-lipoxygenase products on ischemically sensitive abdominal visceral afferents.
The effects of 5- and 15-lipoxygenase products, leukotriene B4 (LTB4) and (8R,15S)-dihydroxyeicosa(5E-9,11,13Z)tetraenoic acid (8R,15S-diHETE), on ischemically sensitive abdominal visceral C fiber afferents were evaluated, because this system is important in sensitizing cutaneous afferents. Single-unit activity of abdominal visceral C fiber afferents was recorded from the right thoracic sympathetic chain of anesthetized cats during 5 min of ischemia. Inhibition of 5-lipoxygenase with WY-50295 tromethamine (5 mg/kg iv) augmented the impulse activity from 0.48 +/- 0.15 to 0.79 +/- 0.24 impulses/s (P < 0.05) in seven ischemically sensitive afferents. Conversely, topical application of LTB4 (125 ng) directly onto the receptive field attenuated impulse activity of 10 ischemically sensitive C fiber afferents from 0.82 +/- 0.23 to 0.42 +/- 0.10 impulses/s (P < 0.05). In additional cats, application of 8R,15S-diHETE (125 ng) onto the receptive field augmented the impulse activity of nine ischemically sensitive C fiber afferents (from 0.48 +/- 0.15 to 0.70 +/- 0.15 impulses/s, P < 0.05) and significantly decreased the mechanical threshold of these nine afferents, whereas application of 8S,15S-diHETE (125 ng), a stereoisomer of 8R,15S-diHETE, attenuated the impulse activity from 0.77 +/- 0.48 to 0.45 +/- 0.13 impulses/s (P < 0.05) in six additional ischemically sensitive C fiber afferents. In animals pretreated with aspirin (50 mg/kg iv, n = 6) or 8S,15S-diHETE (125 ng, n = 6), WY-50295 tromethamine (5 mg/kg iv) still potentiated the impulse activity of ischemically sensitive C fiber afferents. These data indicate that 8R,15S-diHETE interacts with stereospecific receptors to sensitize, whereas LTB4 reduces, the response of abdominal visceral afferents to ischemia. Furthermore the data suggest that the augmented response of afferents to abdominal ischemia after inhibition of 5-lipoxygenase is, at least in part, independent of shunting to the cyclooxygenase or 15-lipoxygenase system.


Navigator channel adaptation to reconstruct three dimensional heart volumes from two dimensional radiotherapy planning data.
Biologically-based models that utilize 3D radiation dosimetry data to estimate the risk of late cardiac effects could have significant utility for planning radiotherapy in young patients. A major challenge arises from having only 2D treatment planning data for patients with long-term follow-up. In this study, we evaluate the accuracy of an advanced deformable image registration (DIR) and navigator channels (NC) adaptation technique to reconstruct 3D heart volumes from 2D radiotherapy planning images for Hodgkin's Lymphoma (HL) patients. Planning CT images were obtained for 50 HL patients who underwent mediastinal radiotherapy. Twelve image sets (6 male, 6 female) were used to construct a male and a female population heart model, which was registered to 23 HL "Reference" patients' CT images using a DIR algorithm, MORFEUS. This generated a series of population-to-Reference patient specific 3D deformation maps. The technique was independently tested on 15 additional "Test" patients by reconstructing their 3D heart volumes using 2D digitally reconstructed radiographs (DRR). The technique involved: 1) identifying a matching Reference patient for each Test patient using thorax measurements, 2) placement of six NCs on matching Reference and Test patients' DRRs to capture differences in significant heart curvatures, 3) adapting the population-to-Reference patient-specific deformation maps to generate population-to-Test patient-specific deformation maps using linear and bilinear interpolation methods, 4) applying population-to-Test patient specific deformation to the population model to reconstruct Test-patient specific 3D heart models. The percentage volume overlap between the NC-adapted reconstruction and actual Test patient's true heart volume was calculated using the Dice coefficient. The average Dice coefficient expressed as a percentage between the NC-adapted and actual Test model was 89.4 ± 2.8%. The modified NC adaptation technique made significant improvements to the population deformation heart models (p = 0.01). As standard evaluation, the residual Dice error after adaptation was comparable to the volumetric differences observed in free-breathing heart volumes (p = 0.62). The reconstruction technique described generates accurate 3D heart models from limited 2D planning data. This development could potentially be used to retrospectively calculate delivered dose to the heart for historically treated patients and thereby provide a better understanding of late radiation-related cardiac effects.


Physically based reflectance model utilizing polarization measurement.
A surface bidirectional reflectance distribution function (BRDF) depends on both the optical properties of the material and the microstructure of the surface and appears as combination of these factors. We propose a method for modeling the BRDF based on a separate optical-property (refractive-index) estimation by polarization measurement. Because the BRDF and the refractive index for precisely the same place can be determined, errors cased by individual difference or spatial dependence can be eliminated. Our BRDF model treats the surface as an aggregation of microfacets, and the diffractive effect is negligible because of randomness. An example model of a painted aluminum plate is presented.


Wheezing and dyspnoea caused by aberrant left innominate artery.
We present a rare case of a branching anomaly of the aortic arch that resulted in wheezing and dyspnoea. The patient was a 60-year-old male with severe wheezing from babyhood, originally diagnosed with severe bronchial asthma. On auscultation, the inspiratory and expiratory wheezes appeared when the patient leaned forward. He also had difficulty in swallowing solid mass. Tests for airway reversibility and hyperresponsiveness were negative, and asthma treatment was ineffective. He had a right aortic arch. A barium oesophagogram and endoscopic examination indicated narrowing of the oesophagus from behind. Three-dimensional reconstruction of enhanced chest CT images indicated a right aortic arch and an aberrant enlarged left innominate artery, which compressed and narrowed the oesophagus and trachea from behind. Although the patient had been diagnosed with intractable bronchial asthma, his symptoms were more likely caused by this mechanical narrowing as wheezing and dyspnoea disappeared completely after total aortic arch replacement operation.


The 2016 Survey of the Association of Directors of Medical Student Education in Psychiatry.
The members of the Association of Directors of Medical Student Education in Psychiatry (ADMSEP) had been last surveyed in 2010 to identify trends in medical student education in psychiatry, teaching methods, academic productivity, and administrative issues. With increasing requirements, ADMSEP members were surveyed in 2016 to characterize current expectations for and needs of medical student educators in psychiatry. A 53-item survey was sent to members of ADMSEP (n=260) and to clerkship directors nationwide. The current survey expanded past surveys with questions about current issues in medical education. Questions included categorical and scaled items (1 = strongly disagree to 5 = strongly agree). All ADMSEP members were invited to complete the survey, but half of the questions were for those who identified as clerkship directors. The survey was sent electronically with two reminders. The survey had a response rate of 42.7%. Half identified themselves as clerkship directors (49.5%), with an average of 6 years as both a clerkship director and member of ADMSEP. Respondents noted that being involved in education (mean 4.1) and ADMSEP (mean 4.2) were a benefit to their career, and also reported that they had adequate access to faculty resources. Respondents also reported concerns with access to educational research consultants and personnel management. Results of the 2016 ADMSEP survey echoed previous membership surveys. However, training in personnel management, diminishing faculty resources, and compensation for education time emerged in this survey. Conducting a membership survey continues to provide beneficial information regarding the administration and education that occurs within psychiatry clerkship. Additionally, topics meriting further details have been identified that will be investigated in future membership surveys.


Subretinal neovascularization after naphthalene damage to the rabbit retina.
To examine the retinotoxic effect of naphthalene, a powerful oxidative agent and a well-known cataractogenic agent. A 10% solution of naphthalene dissolved in paraffin oil was given every other day by gavage to 31 pigmented rabbits for 5 weeks, at a dose of 1 g/kg body weight. Four rabbits who received only paraffin oil served as controls. The eyes were clinically followed up by means of ophthalmoscopy and retinal fluorangiography. At selected intervals, the eyes were examined with light microscopy and transmission electron microscopy. The first lesions were focal and appeared in the periphery of the fundus about 3 weeks after the beginning of treatment and tended to spread over the entire retina. Histologically, there was a degeneration of photoreceptors, accompanied by a reaction and proliferation of retinal pigment epithelium (RPE) that phagocytized the damaged visual cells. After about 3 months, the proliferation of RPE was followed by subretinal neovascularization (SRN). Both mature fenestrated and thick-walled non-fenestrated capillaries penetrated Bruch's membrane, enveloped by abundant fibrous extracellular matrix and accompanied by pericytes. As a consequence of this process, the retina was focally transformed into a "neovascular complex" in which a vascular plexus was intermingled with pseudo-acinar cavities lined by RPE. There were no signs of SRN at retinal fluorangiography, possibly because of the dense microenvironment of extracellular matrix and RPE cells of the neovascular complexes. Naphthalene degeneration of the rabbit retina appears to be a simple model of photoreceptor vulnerability in the first stages of SRN thereafter. The close chronologic and topographic relationship between the appearance of the anomalous vessels and RPE alteration and the close resemblance with previous models of experimental SRN may support the hypothesis of an experimental model of SRN triggered by the RPE.


Process mining routinely collected electronic health records to define real-life clinical pathways during chemotherapy.
There is growing interest in the use of routinely collected electronic health records to enhance service delivery and facilitate clinical research. It should be possible to detect and measure patterns of care and use the data to monitor improvements but there are methodological and data quality challenges. Driven by the desire to model the impact of a patient self-test blood count monitoring service in patients on chemotherapy, we aimed to (i) establish reproducible methods of process-mining electronic health records, (ii) use the outputs derived to define and quantify patient pathways during chemotherapy, and (iii) to gather robust data which is structured to be able to inform a cost-effectiveness decision model of home monitoring of neutropenic status during chemotherapy. Electronic Health Records at a UK oncology centre were included if they had (i) a diagnosis of metastatic breast cancer and received adjuvant epirubicin and cyclosphosphamide chemotherapy or (ii) colorectal cancer and received palliative oxaliplatin and infusional 5-fluorouracil chemotherapy, and (iii) were first diagnosed with cancer between January 2004 and February 2013. Software and a Markov model were developed, producing a schematic of patient pathways during chemotherapy. Significant variance from the assumed care pathway was evident from the data. Of the 535 patients with breast cancer and 420 with colorectal cancer there were 474 and 329 pathway variants respectively. Only 27 (5%) and 26 (6%) completed the planned six cycles of chemotherapy without having unplanned hospital contact. Over the six cycles, 169 (31.6%) patients with breast cancer and 190 (45.2%) patients with colorectal cancer were admitted to hospital. The pathways of patients on chemotherapy are complex. An iterative approach to addressing semantic and data quality issues enabled the effective use of routinely collected patient records to produce accurate models of the real-life experiences of chemotherapy patients and generate clinically useful information. Very few patients experience the idealised patient pathway that is used to plan their care. A better understanding of real-life clinical pathways through process mining can contribute to care and data quality assurance, identifying unmet needs, facilitating quantification of innovation impact, communicating with stakeholders, and ultimately improving patient care and outcomes.


Tissue adhesive wound repair revisited.
The purpose of this experimental study was to compare the effect of a tissue adhesive, N-butyl-2-cyanoacrylate, on the wound's ability to resist infection and gain strength to the effect of percutaneous polypropylene suture. Percutaneous sutures damaged host defenses, inviting the growth of bacteria to a level that was significantly greater than that encountered with the tissue adhesive. Immediately after wound closure, percutaneous sutures provided a more secure closure, as measured by breaking strength, than did tissue adhesives. Seven days later, the breaking strengths of wounds closed by tissue adhesives did not differ significantly from those repaired with percutaneous sutures. Tissue adhesive closure requires less psychomotor skills than suture closure and is accomplished more rapidly than suture closure.


Human Asunder promotes dynein recruitment and centrosomal tethering to the nucleus at mitotic entry.
Recruitment of dynein motors to the nuclear surface is an essential step for nucleus-centrosome coupling in prophase. In cultured human cells, this dynein pool is anchored to nuclear pore complexes through RanBP2-Bicaudal D2 (BICD2) and Nup133- centromere protein F (CENP-F) networks. We previously reported that the asunder (asun) gene is required in Drosophila spermatocytes for perinuclear dynein localization and nucleus-centrosome coupling at G2/M of male meiosis. We show here that male germline expression of mammalian Asunder (ASUN) protein rescues asun flies, demonstrating evolutionary conservation of function. In cultured human cells, we find that ASUN down-regulation causes reduction of perinuclear dynein in prophase of mitosis. Additional defects after loss of ASUN include nucleus-centrosome uncoupling, abnormal spindles, and multinucleation. Coimmunoprecipitation and overlapping localization patterns of ASUN and lissencephaly 1 (LIS1), a dynein adaptor, suggest that ASUN interacts with dynein in the cytoplasm via LIS1. Our data indicate that ASUN controls dynein localization via a mechanism distinct from that of either BICD2 or CENP-F. We present a model in which ASUN promotes perinuclear enrichment of dynein at G2/M that facilitates BICD2- and CENP-F-mediated anchoring of dynein to nuclear pore complexes.


Long-term stability of angle-stable versus conventional locked intramedullary nails in distal tibia fractures.
In the last years intramedullary nailing has become the treatment of choice for most displaced diaphyseal tibia fractures. In contrast intramedullary nailing of distal tibia fractures is accompanied by problems like decreased biomechanical stability. Nevertheless the indications for intramedullary nailing have been extended to include even more distal fractures. The purpose of this study was to compare long-term mechanical characteristics of angle-stable versus conventional locked intramedullary nails in the treatment of unstable distal tibia fractures. Therefore, the effect of time on the mechanical properties of biodegradable sleeves was assessed. 8 pairs of fresh, frozen porcine tibiae were used. The expert tibial nail (Synthes) was equipped with either three conventional locking screws (CL) or the angle-stable locking system (AS), consisting of a special ASLS screw and a biodegradable sleeve. Biomechanical testing included torsional and axial loading at different time-points over 12 weeks. The AS group showed a significantly higher torsional stiffness at all time-points (at least 60%) compared to the CL group (p < 0.001). The neutral zone was at least 5 times higher in the CL group (p < 0.001). The mean axial stiffness was maximum 10% higher (week 6) in the angle-stable locked group compared to the conventional group. There was no significant change of the torsional mechanical characteristics over the 12 weeks in both groups (p > 0.05). For axial stiffness and range of motion significant differences were found in the AS group. The angle-stable locking system (ASLS) with the biodegradable sleeve provides significantly higher long-term stability. Especially the differences determined under torsional loading in this study may have clinical relevance. The ASLS permits the potential to decrease complications like secondary loss of reduction and mal-/non-union.


MRI of the cardiomyopathies.
We examined the potentialities of Magnetic resonance imaging (MRI) in the evaluation of the main cardiomyopathies: hypertrophic, dilated, restrictive and arrhythmogenic right ventricular. The hypertrophic cardiomyopathy is generally adequately investigated by echocardiography, that well defines the myocardial thickening and the obstruction of the left ventricular output. However, by echocardiography we still have difficulties in the evaluation of the apex of the left ventricle and the right ventricle involvement. MRI provides a complete evaluation of the heart with a clear evidence also of the echocardiographic dark zones by means of a clear evidence of the apex of the right ventricle. The dilated form is also well investigated by MRI that provides a clear evaluation of the volumes, mass and ejection fraction by means of the 3D analysis including conditions of the ventricular remodelling. Moreover, this technique helps in the differential diagnosis of acute myocarditis. In the acute phase of myocarditis (first 2 weeks), in fact, the myocardium produces high signal intensity on the T2 weighted sequences due to the presence of oedema. The third form of cardiomyopathy is the restrictive one, characterised by reduced diastolic filling and diastolic volume, normality of the systolic function and parietal thickness, interstitial fibrosis and enlargement of both atria. The mean potentiality of MRI is related to the differential diagnosis with constrictive pericarditis. Only in the former, the pericardium appears irregularly thickened with areas exceeding 4 mm of pericardial thickness. Finally, the right ventricular arrhythmogenic cardiomyopathy represents the main indication to MRI evaluation. With this imaging modality we are can obtain a clear morpho-functional evaluation of the right ventricle and distinguish the intramyocardial adipose substitution characterised by areas of high signal in the myocardium.


Metal-induced DNA translocation leads to DNA polymerase conformational activation.
Binding of the catalytic divalent ion to the ternary DNA polymerase β/gapped DNA/dNTP complex is thought to represent the final step in the assembly of the catalytic complex and is consequently a critical determinant of replicative fidelity. We have analyzed the effects of Mg(2+) and Zn(2+) on the conformational activation process based on NMR measurements of [methyl-(13)C]methionine DNA polymerase β. Unexpectedly, both divalent metals were able to produce a template base-dependent conformational activation of the polymerase/1-nt gapped DNA complex in the absence of a complementary incoming nucleotide, albeit with different temperature thresholds. This conformational activation is abolished by substituting Glu295 with lysine, thereby interrupting key hydrogen bonds necessary to stabilize the closed conformation. These and other results indicate that metal-binding can promote: translocation of the primer terminus base pair into the active site; expulsion of an unpaired pyrimidine, but not purine, base from the template-binding pocket; and motions of polymerase subdomains that close the active site. We also have performed pyrophosphorolysis studies that are consistent with predictions based on these results. These findings provide new insight into the relationships between conformational activation, enzyme activity and polymerase fidelity.


Survival of Prosthodontic Restorations Luted with Resin-Based versus Composite-Based Cements: Retrospective Cohort Study.
The purpose of this study was to evaluate clinical performance, survival, and complications of indirect composite inlays, onlays, and overlays on posterior teeth. Digital records of 282 patients treated between 2014 and 2018 were accessed and analyzed retrospectively. The included patients received 469 composite restorations luted with seven different resin-based types of cement, i.e., Filtek Ultimate Flow, Enamel Plus, Relyx Ultimate, Harvard Premium Flow, Relyx Unicem, Filtek Bulk Fill Flowable, and Filtek Ultimate. The restorations had been clinically and radiographically evaluated annually. The mechanical and clinical complications, e.g., debonding, fracture, and secondary caries, were evaluated and recorded. The examined restorations exhibited a high survival rate (84.9%), and failure was found in only 71 cases. Fracture was the most common cause (n = 36), followed by prosthetic work release (n = 19) and secondary caries (n = 16). There was a statistically significant difference between failure and cement material (Sig. < 0.001); the composite-based cements (87.2%) had a high survival rate compared to the resin-based cement (72.7%). Similarly, the cements with high viscosity (90.2%) had significantly higher survival rates than the low-viscosity cements (78.9%). Moreover, onlays showed higher longevity compared to overlays (Sig. = 0.007), and patients aged under 55 years showed less complications (Sig. = 0.036). Indirect composite restoration was a successful solution to tooth structure loss. The material of the cementation is an important part of the success. Higher survival rate was found in our study when the fixation materials with high viscosity were used, thus suggesting using these materials with indirect restorations. Composite-based cements had significantly higher survival rate than resin-based cements.


[Anti-inflammatory mechanism of Lactobacillus rhamnosus GG in lipopolysaccharide- stimulated HT-29 cell].
Probiotics are live non-pathogenic organisms that belong to the resident microflora, and confer health benefits by multiple mechanisms. Lactobacillus rhamnosus GG (LGG) is one of the probiotic bacteria that ameliorates intestinal injury and inflammation caused by various stimuli. We aimed to evaluate the anti-inflammatory effect and mechanism of LGG in lipopolysaccharide (LPS)-stimulated HT-29 cells. HT-29 cells were stimulated with interleukin (IL)-1β (2 ng/mL), tumor necrosis factor (TNF)-α (20 ng/mL), and LPS (20 μg/mL) in the presence or absence of LGG (10⁷-10⁹ colony forming units/mL). Production of the pro-inflammatory chemokine IL-8 was measured by ELISA and semi-quantitative PCR. Transcriptional activity of NF-kB-responsive gene was evaluated by luciferase assay with reporter gene. Toll-like receptor 4 (TLR4) mRNA expression was assessed by semi-quantitative PCR. The IκBa degradation was evaluated by western blot and intranuclear translocation of NF-κB was determined by western blot and immunofluorescence. LGG did not affect the viability of HT-29 cells. Pretreatment of HT-29 cells with LGG significantly blocked TNF-α, and LPS induced IL-8 activation at both mRNA and protein level (p<0.05). Pretreatment of HT-29 cells with LGG attenuated LPS-induced NF-κB nuclear translocation and also blocked LPS-induced IκBα degradation. LGG also down-regulated TLR4 mRNA activated by LPS. LGG attenuates LPS induced inflammation, and this may be associated with TLR4/NF-κB down-regulation.


Relationship between the functional status scale and the pediatric overall performance category and pediatric cerebral performance category scales.
Functional status assessment methods are important as outcome measures for pediatric critical care studies. To investigate the relationships between the 2 functional status assessment methods appropriate for large-sample studies, the Functional Status Scale (FSS) and the Pediatric Overall Performance Category and Pediatric Cerebral Performance Category (POPC/PCPC) scales. Prospective cohort study with random patient selection at 7 sites and 8 children's hospitals with general/medical and cardiac/cardiovascular pediatric intensive care units (PICUs) in the Collaborative Pediatric Critical Care Research Network. Participants included all PICU patients younger than 18 years. Functional Status Scale and POPC/PCPC scores determined at PICU admission (baseline) and PICU discharge. We investigated the association between the baseline and PICU discharge POPC/PCPC scores and the baseline and PICU discharge FSS scores, the dispersion of FSS scores within each of the POPC/PCPC ratings, and the relationship between the FSS neurologic components (FSS-CNS) and the PCPC. We included 5017 patients. We found a significant (P < .001) difference between FSS scores in each POPC or PCPC interval, with an FSS score increase with each worsening POPC/PCPC rating. The FSS scores for the good and mild disability POPC/PCPC ratings were similar and increased by 2 to 3 points for the POPC/PCPC change from mild to moderate disability, 5 to 6 points for moderate to severe disability, and 8 to 9 points for severe disability to vegetative state or coma. The dispersion of FSS scores within each POPC and PCPC rating was substantial and increased with worsening POPC and PCPC scores. We also found a significant (P < .001) difference between the FSS-CNS scores between each of the PCPC ratings with increases in the FSS-CNS score for each higher PCPC rating. The FSS and POPC/PCPC system are closely associated. Increases in FSS scores occur with each higher POPC and PCPC rating and with greater magnitudes of change as the dysfunction severity increases. However, the dispersion of the FSS scores indicated a lack of precision in the POPC/PCPC system when compared with the more objective and granular FSS. The relationship between the PCPC and the FSS-CNS paralleled the relationship between the FSS and POPC/PCPC system.


The cis-acting replication elements define human enterovirus and rhinovirus species.
Replication of picornaviruses is dependent on VPg uridylylation, which is linked to the presence of the internal cis-acting replication element (cre). Cre are located within the sequence encoding polyprotein, yet at distinct positions as demonstrated for poliovirus and coxsackievirus-B3, cardiovirus, and human rhinovirus (HRV-A and HRV-B), overlapping proteins 2C, VP2, 2A, and VP1, respectively. Here we report a novel distinct cre element located in the VP2 region of the recently reported HRV-A2 species and provide evolutionary evidence of its functionality. We also experimentally interrogated functionality of recently identified HRV-B cre in the 2C region that is orthologous to the human enterovirus (HEV) cre and show that it is dispensable for replication and appears to be a nonfunctional evolutionary relic. In addition, our mutational analysis highlights two amino acids in the 2C protein that are crucial for replication. Remarkably, we conclude that each genetic clade of HRV and HEV is characterized by a unique functional cre element, where evolutionary success of a new genetic lineage seems to be associated with an invention of a novel cre motif and decay of the ancestral one. Therefore, we propose that cre element could be considered as an additional criterion for human rhinovirus and enterovirus classification.


Foci of Schistosomiasis mekongi, Northern Cambodia: II. Distribution of infection and morbidity.
In the province of Kracheh, in Northern Cambodia, a baseline epidemiological survey on Schistosoma mekongi was conducted along the Mekong River between December 1994 and April 1995. The results of household surveys of highly affected villages of the East and the West bank of the river and of school surveys in 20 primary schools are presented. In household surveys 1396 people were examined. An overall prevalence of infection of 49.3% was detected by a single stool examination with the Kato-Katz technique. The overall intensity of infection was 118.2 eggs per gram of stool (epg). There was no difference between the population of the east and west shore of the Mekong for prevalence (P = 0.3) or intensity (P = 0.9) of infection. Severe morbidity was very frequent. Hepatomegaly of the left lobe was detected in 48.7% of the population. Splenomegaly was seen in 26.8% of the study participants. Visible diverted circulation was found in 7.2% of the population, and ascites in 0.1%. Significantly more hepatomegaly (P = 0.001), splenomegaly (P = 0. 001) and patients with diverted circulation (P = 0.001) were present on the west bank of the Mekong. The age group of 10-14 years was most affected. The prevalence of infection in this group was 71.8% and 71.9% in the population of the West and East of the Mekong, respectively. The intensity of infection was 172.4 and 194.2 epg on the West and the East bank, respectively. In the peak age group hepatomegaly reached a prevalence of 88.1% on the west and 82.8% on the east bank. In the 20 schools 2391 children aged 6-16 years were examined. The overall prevalence of infection was 40.0%, ranging from 7.7% to 72.9% per school. The overalls mean intensity of infection was 110.1 epg (range by school: 26.7-187.5 epg). Both prevalence (P = 0.001) and intensity of infection (P = 0.001) were significantly higher in schools on the east side of the Mekong. Hepatomegaly (55.2%), splenomegaly (23.6%), diverted circulation (4. 1%), ascites (0.5%), reported blood (26.7%) and mucus (24.3%) were very frequent. Hepatomegaly (P = 0.001), splenomegaly (P = 0.001), diverted circulation (P = 0.001) and blood in stool (P = 0.001) were significantly more frequent in schools of the east side of the Mekong. Boys suffered more frequently from splenomegaly (P = 0.05), ascites (P = 0.05) and bloody stools (P = 0.004) than girls. No difference in sex was found for the prevalence and intensity of infection and prevalence of hepatomegaly. On the school level prevalence and intensity of infection were highly associated (r = 0. 93, P = 0.0001). The intensity of infection was significantly associated only with the prevalence of hepatomegaly (r = 0.44, P = 0. 05) and blood in stool (r = 0.40, P = 0.02). This comprehensive epidemiological study documents for the first time the public health importance of schistosomiasis mekongi in the Province of Kracheh, Northern Cambodia and points at key epidemiological features of this schistosome species, in particular the high level of morbidity associated with infection.


Subclinical meningoventriculitis as a cause of obstructive hydrocephalus.
Communicating hydrocephalus may complicate infantile bacterial meningitis, typically presenting with systemic features of infection. We report a rare case of 'subclinical meningoventriculitis' causing obstructive hydrocephalus and its challenging management. A healthy 10-week-old immunocompetent male patient presented with failure to thrive and vomiting, secondary to presumed gastro-oesophageal reflux. The child was neurologically alert, afebrile with normal inflammatory markers. Progressive macrocephaly prompted an MRI confirming triventricular hydrocephalus secondary to aqueductal stenosis. An endoscopic third ventriculostomy was performed however abandoned intraoperatively due to the unexpected finding of intraventricular purulent cerebrospinal fluid. A 6-week course of intravenous ceftriaxone was commenced for Escherichia coli meningoventriculitis. However, the child was readmitted 18 days postoperatively with acute hydrocephalus requiring a ventricular washout and staged ventriculoperitoneal shunt insertion at 4 weeks. Serial head circumference measurements are paramount in the assessment of a paediatric patient. In an immunocompetent child, a subclinical fibropurulent meningoventriculitis can result in several management challenges.


Developmental Changes in the ECG of a Hamster Model of Muscular Dystrophy and Heart Failure.
Aberrant autonomic signaling is being increasingly recognized as an important symptom in neuromuscular disorders. The δ-sarcoglycan-deficient BIO TO-2 hamster is recognized as a good model for studying mechanistic pathways and sequelae in muscular dystrophy and heart failure, including autonomic nervous system (ANS) dysfunction. Recent studies using the TO-2 hamster model have provided promising preclinical results demonstrating the efficacy of gene therapy to treat skeletal muscle weakness and heart failure. Methods to accelerate preclinical testing of gene therapy and new drugs for neuromuscular diseases are urgently needed. The purpose of this investigation was to demonstrate a rapid non-invasive screen for characterizing the ANS imbalance in dystrophic TO-2 hamsters. Electrocardiograms were recorded non-invasively in conscious ∼9-month old TO-2 hamsters (n = 10) and non-myopathic F1B control hamsters (n = 10). Heart rate was higher in TO-2 hamsters than controls (453 ± 12 bpm vs. 311 ± 25 bpm, P < 0.01). Time domain heart rate variability, an index of parasympathetic tone, was lower in TO-2 hamsters (12.2 ± 3.7 bpm vs. 38.2 ± 6.8, P < 0.05), as was the coefficient of variance of the RR interval (2.8 ± 0.9% vs. 16.2 ± 3.4%, P < 0.05) compared to control hamsters. Power spectral analysis demonstrated reduced high frequency and low frequency contributions, indicating autonomic imbalance with increased sympathetic tone and decreased parasympathetic tone in dystrophic TO-2 hamsters. Similar observations in newborn hamsters indicate autonomic nervous dysfunction may occur quite early in life in neuromuscular diseases. Our findings of autonomic abnormalities in newborn hamsters with a mutation in the δ-sarcoglycan gene suggest approaches to correct modulation of the heart rate as prevention or therapy for muscular dystrophies.


An alternative strategy for combination therapy: Interactions between polymyxin B and non-antibiotics.
Antimicrobial resistance is increasing and few new antibiotics are in the development pipeline. Alternative strategies to treat infectious diseases, such as combination therapy, are urgently needed. Polymyxin B is a neglected and disused antibiotic with moderate antibacterial activity. In this study, we aimed to find synergistic interactions between polymyxin B and a wide range of non-antibiotics (non-ABs) to improve its efficacy. Thirty non-AB compounds from various drug classes were screened for synergistic potential with sub-minimum inhibitory concentrations (MICs) of polymyxin B in an agar diffusion assay against Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa (3 isolates per species). Potential candidates were further studied in in vitro checkerboard assays, up to 5 isolates per species, using optical density to assess growth. Interactions were assessed with fractional inhibitory concentration index (FICi) analysis and surface response analysis with Loewe, Bliss and Highest Single Agent analysis using the Combenefit program. Twenty non-ABs enhanced polymyxin B activity in the agar diffusion test in one or more species. Of these, three showed a consistent synergistic effect (FICi ≤ 0.5) in the checkerboard assay for at least one species: citalopram, sertraline and spironolactone. Surface response analyses were largely in concordance, and further assessment showed only spironolactone was synergistic with polymyxin B at clinically relevant levels. The screening strategy used showed consistent synergism in vitro between polymyxin B and some non-ABs for A. baumannii, E. coli and K. pneumoniae. The synergistic interactions found merit further exploration as alternative strategies for difficult-to-treat infections.


Selectivity of lingual nerve fibers to chemical stimuli.
The cell bodies of the lingual branch of the trigeminal nerve were localized in the trigeminal ganglion using extracellular recordings together with horseradish peroxidase labeling from the tongue. Individual lingual nerve fibers were characterized with regard to their conduction velocities, receptive fields, and response to thermal, mechanical, and chemical stimuli. Fibers were classified as C, A delta, A beta, cold, and warm. The chemical stimuli included NaCl, KCl, NH4Cl, CaCl2, menthol, nicotine, hexanol, and capsaicin. With increasing salt concentration the latency of the response decreased and the activity increased. The responses elicited by salts (to 2.5 M), but not nonpolar stimuli such as menthol, were reversibly inhibited by 3.5 mM of the tight junction blocker, LaCl3. These data suggest that salts diffuse into stratified squamous epithelia through tight junctions in the stratum corneum and stratum granulosum, whereupon they enter the extracellular space. 11 C fibers were identified and 5 were characterized as polymodal nociceptors. All of the C fibers were activated by one or more of the salts NaCl, KCl, or NH4Cl. Three C fibers were activated by nicotine (1 mM), but none were affected by CaCl2 (1 M), menthol (1 mM), or hexanol (50 mM). However, not all C fibers or even the subpopulation of polymodals were activated by the same salts or by nicotine. Thus, it appears that C fibers display differential responsiveness to chemical stimuli. A delta fibers also showed differential sensitivity to chemicals. Of the 35 characterized A delta mechanoreceptors, 8 responded to NaCl, 9 to KCl, 9 to NH4Cl, 0 to CaCl2, menthol, or hexanol, and 2 to nicotine. 8 of 9 of the cold fibers (characterized as A delta's) responded to menthol, none responded to nicotine, 8 of 16 were inhibited by hexanol, 9 of 19 responded to 2.5 M NH4Cl, 5 of 19 responded to 2.5 M KCl, and 1 of 19 responded to 2.5 M NaCl. In summary, lingual nerve fibers exhibit responsiveness to chemicals introduced onto the tongue. The differential responses of these fibers are potentially capable of transmitting information regarding the quality and quantity of chemical stimuli from the tongue to the central nervous system.


Reversible left ventricular apical ballooning associated with jet lag in a Taiwanese woman: A case report.
Reversible left ventricular apical ballooning, without coronary artery stenosis, is a novel heart syndrome mimicking acute myocardial infarction, and is very rare in Taiwan. A 74-year-old Taiwanese woman returned from travelling abroad for one week and suffered from persistent, severe jet lag with sleep disturbance. She had a cold exacerbated by bronchial asthma for three days. She presented with sudden onset of chest pain after drinking three cups of coffee and taking a sauna for more than 1 h. On admission, an electrocardiogram showed ST segment elevation in leads II, III, aVF and V(3-6), and cardiac enzyme tests revealed minimal elevation. An echocardiogram showed apical ballooning and basal hyperkinesias of the left ventricle (LV) in systole. A coronary angiogram on the second day was normal, while the ST segment was still elevated, and the patient continued to experience chest pain. A negative T wave developed three days later. The electrocardiogram abnormality and LV dysfunction resolved completely six months later. A takotsubo (ampulla) cardiomyopathy was diagnosed. The activated myocardial adrenergic nervous system, stimulated by acute and marked stress in this patient, with more adrenergic innervations distributed in the apex of the LV, may be the trigger for this novel cardiac syndrome.


Impact of a Women-focused Professional Organization on Academic Retention and Advancement: Perceptions From a Qualitative Study.
Organizations to promote career networking and mentorship among women are recommended as a best practice to support the recruitment and retention of women physicians; however, the impact of such organizations is unknown. Our primary objective is to describe the impact of a national woman-focused organization for academic emergency physicians on retention and advancement. We conducted semistructured interviews of past and present organization leaders, as well as members at varying stages in their careers. Physicians with experience in qualitative methods conducted interviews and coded all transcripts using inductive content analysis techniques. Themes were reviewed and discussed to ensure consensus. We performed 17 interviews lasting 20 to 30 minutes each, resulting in 476 total minutes of transcript. Participants represented varying stages of career experience, ranging from 2 to 35 years since residency completion (median = 9.5 years). Median years of participation in the woman-focused organization was 10 years. Over half (53%) of participants were past presidents of the organization. The dominant themes encompassed facilitating academic advancement through scholarly productivity, leadership experiences, awards, and promotions; mentorship and sponsorship; peer support and collaborations; reduced professional isolation; and initiatives to address systemic gender inequities and challenges, including strategies to navigate bias, promote pay equity, and advocate for family-friendly workplace policies. Active participation in a woman-focused professional organization enhances members' career retention and advancement by creating opportunities and relationships that facilitate leadership, enabling scholarly work to advance equity and inclusion, and cultivating a sense of belonging. While challenges and barriers persist, the myriad benefits of a women-focused professional organization reported by members and leaders represent important steps toward greater equity for women and other underrepresented groups in academic medicine.


Correlates of bushmeat in markets and depletion of wildlife.
We used data on number of carcasses of wildlife species sold in 79 bushmeat markets in a region of Nigeria and Cameroon to assess whether species composition of a market could be explained by anthropogenic pressures and environmental variables around each market. More than 45 mammal species from 9 orders were traded across all markets; mostly ungulates and rodents. For each market, we determined median body mass, species diversity (game diversity), and taxa that were principal contributors to the total number of carcasses for sale (game dominance). Human population density in surrounding areas was significantly and negatively related to the percentage ungulates and primates sold in markets and significantly and positively related to the proportion of rodents. The proportion of carnivores sold was higher in markets with high human population densities. Proportion of small-bodied mammals (<1 kg) sold in markets increased as human population density increased, but proportion of large-bodied mammals (>10 kg) decreased as human population density increased. We calculated an index of game depletion (GDI) for each market from the sum of the total number of carcasses traded per annum and species, weighted by the intrinsic rate of natural increase (rmax ) of each species, divided by individuals traded in a market. The GDI of a market increased as the proportion of fast-reproducing species (highest rmax ) increased and as the representation of species with lowest rmax (slow-reproducing) decreased. The best explanatory factor for a market's GDI was anthropogenic pressure-road density, human settlements with >3000 inhabitants, and nonforest vegetation. High and low GDI were significantly differentiated by human density and human settlements with >3000 inhabitants. Our results provided empirical evidence that human activity is correlated with more depleted bushmeat faunas and can be used as a proxy to determine areas in need of conservation action.


CD28.OX40 co-stimulatory combination is associated with long in vivo persistence and high activity of CAR.CD30 T-cells.
The prognosis of many patients with chemotherapy-refractory or multiply relapsed CD30+ non-Hodgkin Lymphoma (NHL) or Hodgkin lymphoma (HL) still remains poor, and novel therapeutic approaches are warranted to address this unmet clinical need. In light of this consideration, we designed and pre-clinically validated a Chimeric Antigen Receptor (CAR) construct characterized by a novel anti-CD30 single-chain variable-fragment cassette, linked to CD3ζ by the signaling domains of two costimulatory molecules, namely either CD28.4-1BB or CD28.OX40. We found that CAR.CD30 T-cells exhibit remarkable cytolytic activity in vitro against HL and NHL cell lines, with sustained proliferation and pro-inflammatory cytokine production, even after multiple and sequential lymphoma cell challenges. CAR.CD30 T-cells also demonstrated anti-lymphoma activity in two in vivo xenograft immune-deficient mouse models of metastatic HL and NHL. We observed that administration of CAR.CD30 T-cells, incorporating the CD28.OX40 costimulatory domains and manufactured in the presence of IL7 and IL15, were associated with the best overall survival in the treated mice, along with the establishment of a long-term immunological memory, able to protect mice from further tumor re-challenge. Our data indicate that, in the context of in vivo systemic metastatic xenograft mouse models, the costimulatory machinery of CD28.OX40 is crucial for improving persistence, in vivo expansion and proliferation of CAR.CD30 T-cells upon tumor encounter. CD28.OX40 costimulatory combination is ultimately responsible for the antitumor efficacy of the approach, paving the way to translate this therapeutic strategy in patients with CD30+ HL and NHL.


Within-subject joint independent component analysis of simultaneous fMRI/ERP in an auditory oddball paradigm.
The integration of event-related potential (ERP) and functional magnetic resonance imaging (fMRI) can contribute to characterizing neural networks with high temporal and spatial resolution. This research aimed to determine the sensitivity and limitations of applying joint independent component analysis (jICA) within-subjects, for ERP and fMRI data collected simultaneously in a parametric auditory frequency oddball paradigm. In a group of 20 subjects, an increase in ERP peak amplitude ranging 1-8 μV in the time window of the P300 (350-700 ms), and a correlated increase in fMRI signal in a network of regions including the right superior temporal and supramarginal gyri, was observed with the increase in deviant frequency difference. JICA of the same ERP and fMRI group data revealed activity in a similar network, albeit with stronger amplitude and larger extent. In addition, activity in the left pre- and post-central gyri, likely associated with right hand somato-motor response, was observed only with the jICA approach. Within-subject, the jICA approach revealed significantly stronger and more extensive activity in the brain regions associated with the auditory P300 than the P300 linear regression analysis. The results suggest that with the incorporation of spatial and temporal information from both imaging modalities, jICA may be a more sensitive method for extracting common sources of activity between ERP and fMRI.


Counseling in fetal medicine: agenesis of the corpus callosum.
In this Review, we aim to provide up-to-date and evidence-based answers to common questions regarding the diagnosis and prognosis of prenatally detected agenesis of the corpus callosum (ACC). A systematic literature search was performed to identify all reports of ACC and reference lists of articles were identified. ACC involves partial or complete absence of the main commissural pathway that connects the two cerebral hemispheres, and can be isolated (with no other abnormalities) or complex (coexisting with other abnormalities). It is a rare finding and the prevalence is difficult to estimate because of selection bias in reported series. The corpus callosum (CC) can be assessed on ultrasound by direct visualization, but indirect features, such as ventriculomegaly, absence of the cavum septi pellucidi or widening of interhemispheric fissure, are often the reason for detection in a screening population. Careful imaging in a center with a high level of expertise is required to make a full assessment and to exclude coexisting abnormalities, which occur in about 46% of fetuses. When available, magnetic resonance imaging appears to be an important adjunct as it allows direct visualization. It can reduce false-positive rates on ultrasound and can confirm ACC, it can assess whether this is complete or partial and it can help in detecting coexisting brain abnormalities not seen on ultrasound. The overall rate of chromosomal abnormality in fetuses with ACC is 18%, but this high rate includes both isolated and complex ACC; more recent studies suggest that chromosomal abnormalities are rare in isolated cases. Nevertheless, postnatal follow-up studies suggest that about 15% of cases thought to be isolated prenatally were found to have associated abnormalities after birth. Neurodevelopmental outcome in isolated ACC was recently reported in a systematic review and suggested normal outcome in about 65-75% of cases. Findings need to be considered in light of the several limitations of existing studies, in terms of study design, selection bias, varying definitions and imaging protocols, ascertainment bias and lack of control groups. These uncertainties mean that antenatal counseling is difficult and further large prospective studies are needed.


Self-assembled uracil complexes containing tautomeric uracils: an IRMPD spectroscopic and computation study of the structures of gaseous uracilnCa2+ (n = 4, 5, or 6) complexes.
The structures of doubly-charged uracil (U) complexes with Ca2+, UnCa2+ (n = 4, 5, 6), were studied by infrared multiple photon dissociation (IRMPD) spectroscopy and computational methods. The ions were produced by electrospray ionization (ESI) and were isolated in the gas phase in a Fourier-transform ion cyclotron resonance mass spectrometer (FT-ICR-MS). The recorded IRMPD spectra in both the fingerprint and the C-H/N-H/O-H stretching regions, combined with computed vibrational spectra, reveal that the structures present in the greatest abundance consist of both canonical uracil as well as the lactam (or colloquially "enol") tautomer of uracil. U4Ca2+ consists of two hydrogen-bonded dimers of uracil, one canonical and one tautomer, with each uracil interacting with Ca2+ through a carbonyl oxygen. The structures most consistent with the vibrational spectrum of U6Ca2+ consist of two hydrogen-bonded uracil trimers, each composed of two canonical and one enolic uracil, with each uracil also interacting with Ca2+ through carbonyl oxygen. U5Ca2+ consists of one of the aforementioned trimers and dimers, each containing one enol tautomerized uracil. The computed structures whose vibrational spectra best agree with the experimental vibrational spectra are also the lowest-energy structures for all three complexes. This study clearly shows that some uracils adopt the normally very high energy enol tautomer in the lowest energy gas phase complexes of uracil with a doubly-charged ion like Ca2+.


RNA sequencing identifies a novel USP9X-USP6 promoter swap gene fusion in a primary aneurysmal bone cyst.
Primary aneurysmal bone cyst (ABC) is a benign multiloculated cystic lesion of bone that is defined cytogenetically by USP6 gene rearrangements. Rearrangements involving USP6 are promoter swaps, usually generated by fusion of the noncoding upstream exons of different partner genes with exon 1 or 2 of USP6, thus leading to transcriptional upregulation of full-length USP6 coding sequence. Testing for USP6 rearrangements is used diagnostically to distinguish it from secondary ABC and other giant cell-rich primary bone tumors. In this report, we present a case of a 16-year-old male with a primary ABC of the left distal femur. USP6 break apart fluorescence in situ hybridization was positive for a rearrangement and conventional chromosome analysis identified a reciprocal X;17 translocation. In order to identify the putative USP6 fusion partner, we performed RNA sequencing and uncovered a novel USP9X-USP6 promoter swap fusion. This result was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and by mate pair sequencing thus showing the utility of these alternative methodologies in identifying novel fusion candidates. Ubiquitin-specific protease 9X (USP9X), like USP6, encodes a highly conserved substrate-specific deubiquitylating enzyme. USP9X is highly expressed in a number of tissue types and acts as both an oncogene and tumor suppressor in several human cancers. We conclude that oncogenic activation of USP6 via USP9X promoter exchange represents a novel driver of primary ABC formation.


Perceptions of service user and carer involvement in healthcare education and impact on students' knowledge and practice: a literature review.
The involvement of service users in healthcare education has followed an increasing focus on patient-centred services. CETL4HealthNE, regional healthcare education collaboration, undertook a literature review to inform involvement strategies. To identify--approaches used to involve service users in healthcare education curricula; perceptions of key stakeholders; impact of involvement on students' knowledge and skills, and the quality of care delivered. A structured search of the literature on service user involvement in Higher Education healthcare curricular activity was undertaken (July 2006-February 2007). Papers were screened and data extracted and synthesized according to the aspect of the curriculum enhanced by involvement and level of impact evaluation. Thirty papers addressed interventions in pre- and 11 in post-registration education. Thirty studies reported on evaluation. Students and service users both benefited from service user involvement in health care programmes. There was limited evidence that involvement changed behaviour in practice or significantly benefited the recipient of care. Service user involvement in curricular developments for healthcare professionals is a policy imperative aimed at improving their experience of services. Despite a limited and weak traditional evidence base for impact on students' knowledge and practice, both students and service users identify benefits from engagement. Further development of evaluation methodologies is required.


Endovascular embolization of cervical hemangiopericytoma with Onyx-18: case report and review of the literature.
Cervical hemangiopericytoma (HPC) is a rare tumor involving dura and bone. Because HPC shares some imaging features with meningioma, hemangioblastoma, schwannoma and solitary fibrous tumors; histology with appropriate immunohistochemistry is essential for its diagnosis and treatment. HPC is highly vascular and has a high rate of local recurrence following resection. Thus it can pose significant therapeutic challenges. To the best of our knowledge this is the 13th reported case of HPC of the cervical spine and the first case of cervical HPC treated using embolization with Onyx-18. In addition, a systematic review of the literature is presented describing previous experience with HPC of the cervical region. The case of a 61-year-old woman is reported who presented with a 7 month history of dizziness, mild ataxia and left-sided tinnitus when supine. MRI of the cervical spine demonstrated a gadolinium enhancing mass arising from the dorsal cervical dura, extending from the occiput to C2, with internal flow voids and extension into the C2 lamina and spinous process. Digital subtraction angiography demonstrated multiple corkscrew-shaped supplying vessels, marked hypervascularity, rapid arteriovenous shunting and delayed contrast washout. Super selective injection of Onyx-18 yielded approximately 90% embolization of the lesion, followed by subtotal resection and radiotherapy. Histology confirmed the presumptive diagnosis of HPC. Preoperative embolization with Onyx-18 should be considered for large HPC as it may decrease the risk of intraoperative hemorrhage. In this case, embolization with Onyx-18 did not preclude histopathologic diagnosis of HPC.


AUSOM: a 3D placement guide for orthodontic mini-implants.
Anchorage management is an important issue in orthodontic treatment and plays a big role in the success or failure of orthodontic treatment. Using a mini-implant system as skeletal anchorage is a suitable solution if maximum or absolute anchorage control is required. Applying mini-implants requires pinpointing their optimal position to prevent damaging adjacent anatomical structures. There are three considerations in locating proper mini-implant position: the point of implant insertion, the angle of implant insertion in the anterior-posterior direction, and the angulation of implant insertion in the vertical plane. Here, we illustrate a new three-dimensional mini-implant guide (Aleppo University Surgical Orthodontic Miniscrew Guide [AUSOM]) developed for determination of ideal mini-implant placement and demonstrate AUSOM's role in mini-implant success. After reviewing of articles about mini-implants in orthodontic treatment, we determined the characteristics of an ideal orthodontic implant guide (IOIG). We invented a new instrument, AUSOM, that consists of four components: a horizontal part, a vertical part, a graduation guide, and film-holding part. Forty mini-implants were inserted for orthodontic treatment purposes; AUSOM and wire guides were used to insert the mini-implants. We found AUSOM to be a practical and accurate device. It works as a radiographic-locating device and a mini-implant surgical placement guide. The failure rate of mini-implants placed by AUSOM was lower than that of those placed by simple metallic guides. Clinical randomized studies must be carried out to prove AUSOM's role in improving the success rate of mini-implant placement.


Differential contributions of the left and right inferior parietal lobules to number processing.
We measured cerebral activation with functional magnetic resonance imaging at 3 Tesla while eight healthy volunteers performed various number processing tasks known to be dissociable in brain-lesioned patients: naming, comparing, multiplying, or subtracting single digits. The results revealed the activation of a circuit comprising bilateral intraparietal, prefrontal, and anterior cingulate components. The extension and lateralization of this circuit was modulated by task demands. The intraparietal and prefrontal activation was more important in the right hemisphere during the comparison task and in the left hemisphere during the multiplication task and was intensely bilateral during the subtraction task. Thus, partially distinct cerebral circuits with the dorsal parietal pathway underlie distinct arithmetic operations.


Understanding the benign nature of SIV infection in natural hosts.
In striking contrast to HIV infection, natural SIV infection of African nonhuman primates is asymptomatic and usually does not induce significant CD4+ T cell depletion despite high levels of virus replication. Recently, significant progress has been made in understanding the mechanisms underlying the remarkable difference in infection outcome between natural and nonnatural HIV/SIV hosts. These advances include the identification of limited immune activation as a key factor protecting natural SIV hosts from AIDS and the discovery of low CC chemokine receptor 5 expression on CD4+ T cells as a specific and consistent immunologic feature in these animals. Further elucidation of the pathways by which the differences in immune activation between natural and nonnatural hosts are manifest holds promise for the design of novel therapeutic approaches to HIV infection.


Mikulicz's disease: a new perspective and literature review.
To report the clinical and pathophysiologic features of two patients with Mikulicz's disease and to further characterize recommendations for diagnosis and management with a review of the literature. Retrospective nonrandomized consecutive case series, Jules Stein Eye Institute, David Geffen School of Medicine at UCLA. Mikulicz's disease is characterized by symmetric lacrimal, parotid, and submandibular gland enlargement with associated lymphocytic infiltrations. The authors noted two cases of Mikulicz's disease. The diagnosis of Mikulicz's disease was based on the following criteria: 1) symmetric and persistent swelling of the lacrimal glands and either or both of the major salivary glands (parotid and submandibular); and 2) the exclusion of other diseases that may mimic this presentation, such as sarcoidosis, viral infection, or lymphoproliferative disorders. Mikulicz's disease is a condition in which there is bilateral lacrimal and salivary gland swelling that is not associated with other systemic conditions. The condition is self-limiting and most often, the diagnosis is a clinical one. Previously, Mikulicz's disease was often considered as a subtype of Sjögren's syndrome (SS). Clinical and immunologic differences between Mikulicz's disease and SS may warrant further consideration of Mikulicz's disease as a specific autoimmune phenomenon separate from SS, and Mikulicz's disease may be amenable to different treatment modalities than those employed in patients with SS.