Protease inhibitor-based triple therapy in chronic hepatitis C: guidelines by the French Association for the Study of the Liver.
The recent marketing authorizations and hence availability of the new protease inhibitors, telaprevir and boceprevir, have profoundly changed the management of chronic hepatitis C patients. Guidelines for the use of these new drugs as part of a triple therapy, in combination with the standard therapy of peginterferon plus ribavirin, are proposed. The guidelines have been drawn up and evaluated by a meeting of the French Association for the Study of the Liver, posted online for comments, and extensively reviewed by international experts. The current published data on the various treatment strategies are reviewed. The guidelines address the majority of patient profiles including treatment-naïve patients and patients with failure of previous treatment. They recommend which patients should be treated with triple therapy and consider the results of triple therapy including the factors that are predictive of response. They consider the circumstances in which the length of triple therapy can be shortened and the advantages of a peginterferon plus ribavirin lead-in phase. Virological monitoring, stopping criteria, the evaluation of resistance to protease inhibitors, practical treatment management, treatment adherence and the management of side effects are discussed and simple guidelines proposed.


Surveying capsid assembly pathways through simulation-based data fitting.
Virus capsid assembly has attracted considerable interest from the biophysical modeling community as a model system for complicated self-assembly processes. Simulation methods have proven valuable for characterizing the space of possible kinetics and mechanisms of capsid assembly, but they have so far been able to say little about the assembly kinetics or pathways of any specific virus. It is not possible to directly measure the detailed interaction rates needed to parameterize a model, and there is only a limited amount of experimental evidence available to constrain possible pathways, with almost all of it gathered from in vitro studies of purified coat proteins. In prior work, we developed methods to address this problem by using simulation-based data-fitting to learn rate parameters consistent with both structure-based rule sets and experimental light-scattering data on bulk assembly progress in vitro. We have since improved these methods and extended them to fit simulation parameters to one or more experimental light-scattering curves. Here, we apply the improved data-fitting approach to three capsid systems-human papillomavirus (HPV), hepatitis B virus (HBV), and cowpea chlorotic mottle virus (CCMV)-to assess both the range of pathway types the methods can learn and the diversity of assembly strategies in use between these viruses. The resulting fits suggest three different in vitro assembly mechanisms for the three systems, with HPV capsids fitting a model of assembly via a nonnucleation-limited pathway of accumulation of individual capsomers while HBV and CCMV capsids fit similar but subtly different models of nucleation-limited assembly through ensembles of pathways involving trimer-of-dimer intermediates. The results demonstrate the ability of such data fitting to learn very different pathway types and show some of the versatility of pathways that may exist across real viruses.


Similar Clinical Severity and Outcomes for Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Pediatric Musculoskeletal Infections.
Prior studies of pediatric musculoskeletal infection have suggested that methicillin-resistant Staphylococcus aureus (MRSA) infections result in worse outcomes compared with infections with methicillin-susceptible S aureus (MSSA) strains. Based on these results, clinical prediction algorithms have been developed to differentiate between MRSA and MSSA early in a patient's clinical course. This study compares hospital outcomes for pediatric patients with MRSA and MSSA musculoskeletal infection presenting to the emergency department at a large tertiary care children's hospital. A retrospective study identified pediatric patients with S aureus musculoskeletal infection over a 5-year period (2008-2013) by sequential review of all pediatric orthopedic consults. Relevant demographic information, laboratory values, and clinical outcomes were obtained from the electronic medical record. Of the 91 identified cases of S aureus pediatric musculoskeletal infection, there were 49 cases of MRSA infection (53%) and 42 cases of MSSA infection (47%). There were no significant differences between MRSA and MSSA infections in median hospital length of stay (4.8 vs 5.7 days, P = .50), febrile days (0.0 vs 1.5 days, P = .10), and antibiotic duration (28 vs 34 days, P = .18). Methicillin-resistant S aureus infections were more likely to require operative intervention than MSSA infection (85% vs 62%, P = .15). A logistic regression model based on C-reactive protein, temperature, white blood cell count, pulse, and respiratory rate at presentation demonstrated poor ability to differentiate between MRSA and MSSA infection. The results demonstrated no significant differences between MSSA and MRSA musculoskeletal infections for most hospital outcomes measured. However, MRSA infections required more operative interventions than MSSA infections. In addition, a predictive model based on severity markers obtained at presentation was unable to effectively differentiate between MRSA and MSSA infection. The clinical utility and capacity for early differentiation of MRSA and MSSA depends on virulence patterns that may vary temporally and geographically.


Simultaneous localization of lumbar vertebrae and intervertebral discs with SVM-based MRF.
This paper presents a method for localizing and labeling the lumbar vertebrae and intervertebral discs in mid-sagittal MR image slices. The approach is based on a Markov-chain-like graphical model of the ordered discs and vertebrae in the lumbar spine. The graphical model is formulated by combining local image features and semiglobal geometrical information. The local image features are extracted from the image by employing pyramidal histogram of oriented gradients (PHOG) and a novel descriptor that we call image projection descriptor (IPD). These features are trained with support vector machines (SVM) and each pixel in the target image is locally assigned a score. These local scores are combined with the semiglobal geometrical information like the distance ratio and angle between the neighboring structures under the Markov random field (MRF) framework. An exact localization of discs and vertebrae is inferred from the MRF by finding a maximum a posteriori solution efficiently using dynamic programming. As a result of the novel features introduced, our system can scale-invariantly localize discs and vertebra at the same time even in the existence of missing structures. The proposed system is tested and validated on a clinical lumbar spine MR image dataset containing 80 subjects of which 64 have disc- and vertebra-related diseases and abnormalities. The experiments show that our system is successful even in abnormal cases and our results are comparable to the state of the art.


Model-Based Angular Scan Error Correction of an Electrothermally-Actuated MEMS Mirror.
In this paper, the actuation behavior of a two-axis electrothermal MEMS (Microelectromechanical Systems) mirror typically used in miniature optical scanning probes and optical switches is investigated. The MEMS mirror consists of four thermal bimorph actuators symmetrically located at the four sides of a central mirror plate. Experiments show that an actuation characteristics difference of as much as 4.0% exists among the four actuators due to process variations, which leads to an average angular scan error of 0.03°. A mathematical model between the actuator input voltage and the mirror-plate position has been developed to predict the actuation behavior of the mirror. It is a four-input, four-output model that takes into account the thermal-mechanical coupling and the differences among the four actuators; the vertical positions of the ends of the four actuators are also monitored. Based on this model, an open-loop control method is established to achieve accurate angular scanning. This model-based open loop control has been experimentally verified and is useful for the accurate control of the mirror. With this control method, the precise actuation of the mirror solely depends on the model prediction and does not need the real-time mirror position monitoring and feedback, greatly simplifying the MEMS control system.


Separate mechanisms of ion oligomerization tune the physicochemical properties of n-butylammonium acetate: cation-base clusters vs. anion-acid dimers.
We investigated the ability of the ions comprising protic ionic liquids to strongly interact with their neutral acid and base forms through the characterization of n-butylammonium acetate ([C4NH3][OAc]) in the presence of excess n-butylamine (C4NH2) or excess acetic acid (HOAc). The conjugate and parent acid or base form new nonstoichiometric, noncovalently bound species (i.e., oligomeric ions) which change the physical and chemical properties of the resulting liquids, thus offering tunability. The effects of adding C4NH2 or HOAc to [C4NH3][OAc] on the resulting thermal and spectroscopic properties differ and suggest that C4NH2 interacts primarily with [C4NH3]+ to form 3-dimensional polymeric networks likely similar to those in H2O/[H3O]+, while HOAc interacts primarily with [OAc]- to form oligomeric ions (e.g., [H(OAc)2]-). The densities of the systems increased with the increase of acid content and reached a maximum when the acid molar fraction was 0.90, but decreased with increasing amine concentration. The viscosities decreased significantly with increasing acid or base concentration. The solvent properties of the mixtures were assessed by measuring the solubilities of benzene, ethyl acetate, diethyl ether, heptane, ibuprofen free acid, and lidocaine free base. The solubilities of the organic solutes and active pharmaceutical ingredients can be tuned with the concentration of acid or amine in the mixtures. In addition, crystallization of the active pharmaceutical ingredients can be induced with the modification of the composition of the mixtures. These observations support the usage of these mixtures for the synthesis and purification of acid or basic active pharmaceutical ingredients in the pharmaceutical industry.


Computational identification of insertional mutagenesis targets for cancer gene discovery.
Insertional mutagenesis is a potent forward genetic screening technique used to identify candidate cancer genes in mouse model systems. An important, yet unresolved issue in the analysis of these screens, is the identification of the genes affected by the insertions. To address this, we developed Kernel Convolved Rule Based Mapping (KC-RBM). KC-RBM exploits distance, orientation and insertion density across tumors to automatically map integration sites to target genes. We perform the first genome-wide evaluation of the association of insertion occurrences with aberrant gene expression of the predicted targets in both retroviral and transposon data sets. We demonstrate the efficiency of KC-RBM by showing its superior performance over existing approaches in recovering true positives from a list of independently, manually curated cancer genes. The results of this work will significantly enhance the accuracy and speed of cancer gene discovery in forward genetic screens. KC-RBM is available as R-package.


Molecular cloning of the gene for the human placental GTP-binding protein Gp (G25K): identification of this GTP-binding protein as the human homolog of the yeast cell-division-cycle protein CDC42.
We have isolated cDNA clones from a human placental library that code for a low molecular weight GTP-binding protein originally designated Gp (also called G25K). This identification is based on comparisons with the available peptide sequences for the purified human Gp protein and the use of two highly specific anti-peptide antibodies. The predicted amino acid sequence of the protein is very similar to those of various members of the ras superfamily of low molecular weight GTP-binding proteins, including the N-, Ki-, and Ha-ras proteins (30-35% identical), the rho proteins (approximately 50% identical), and the rac proteins (approximately 70% identical). The highest degree of sequence identity (80%) is found with the Saccharomyces cerevisiae cell-division-cycle protein CDC42. The human placental gene, which we designate CDC42Hs, complements the cdc42-1 mutation in S. cerevisiae, which suggests that this GTP-binding protein is the human homolog of the yeast protein.


High-frequency repetitive transcranial magnetic stimulation in schizophrenia: a combined treatment and neuroimaging study.
Repetitive transcranial magnetic stimulation (rTMS) of frontal brain regions is under study as a non-invasive method in the treatment of affective disorders. Recent publications provide increasing evidence that rTMS may be useful in treating schizophrenia. Results are most intriguing, demonstrating a reduction of negative symptoms following high-frequency rTMS. In this context, disentangling of negative and depressive symptoms is of the utmost importance when understanding specific rTMS effects on schizophrenic symptoms. Using a sham-controlled parallel design, 20 patients with schizophrenia were included in the study. Patients were treated with high-frequency 10 Hz rTMS over 10 days. Besides clinical ratings, ECD-SPECT (technetium-99 bicisate single photon emission computed tomography) imaging was performed before and after termination of rTMS treatment. High-frequency rTMS leads to a significant reduction of negative symptoms combined with a trend for non-significant improvement of depressive symptoms in the active stimulated group as compared with the sham stimulated group. Additionally, a trend for worsening of positive symptoms was observed in the actively treated schizophrenic patients. In both groups no changes in regional cerebral blood flow could be detected by ECD-SPECT. Beneficial effects of high-frequency rTMS on negative and depressive symptoms were found, together with a trend for worsening positive symptoms in schizophrenic patients.


Long- and short-lived murine hematopoietic stem cell clones individually identified with retroviral integration markers.
The proliferative longevity of totipotent hematopoietic stem cells (THSC) is a limiting factor in normal hematopoiesis and in therapy by cell- or gene-replacement, but has not yet been ascertained. We have followed the long-term fate of individual clones of mouse THSC from fetal liver or adult bone marrow, after labeling in culture, followed by engraftment and serial transplantation in unirradiated W/Wv-C57BL/6 hosts. The ancestor cell of each clone and its mitotic progeny were uniquely identifiable retrospectively by the DNA integration pattern experimentally produced by replication-incompetent recombinant murine retroviruses. These viruses provided physiologically neutral markers. The marked clones proved to be derived from THSC, based on their contributions to a wide array of myeloid and lymphoid blood lineages in the hosts. The label also identified the target cells as the population displaying clonal succession. The various labeled stem cell clones proliferated for substantially different periods of time. The longest observed clone endured, after the original cell was marked, for at least 2 1/2 years--the equivalent of a mouse's lifetime. However, the results suggest that THSC clones are not all long-lived and that even the longest-lived ones may not be potentially immortal. Thus, the unpredictable lifespan of any given THSC clone indicates the desirability of introducing multiple clones in therapeutic transplants.


Active room compensation for sound reinforcement using sound field separation techniques.
This work investigates how the sound field created by a sound reinforcement system can be controlled at low frequencies. An indoor control method is proposed which actively absorbs the sound incident on a reflecting boundary using an array of secondary sources. The sound field is separated into incident and reflected components by a microphone array close to the secondary sources, enabling the minimization of reflected components by means of optimal signals for the secondary sources. The method is purely feed-forward and assumes constant room conditions. Three different sound field separation techniques for the modeling of the reflections are investigated based on plane wave decomposition, equivalent sources, and the Spatial Fourier transform. Simulations and an experimental validation are presented, showing that the control method performs similarly well at enhancing low frequency responses with the three sound separation techniques. Resonances in the entire room are reduced, although the microphone array and secondary sources are confined to a small region close to the reflecting wall. Unlike previous control methods based on the creation of a plane wave sound field, the investigated method works in arbitrary room geometries and primary source positions.


Development of large granular lymphocytes with anomalous, nonspecific cytotoxicity in clones derived from Ly-2+ T cells.
T cells cultured at limit dilution for 8 days in a concanavalin A-stimulated, filler-cell and growth factor-supported system produced cytolytic clones with high efficiency. These clones were not specific, lysing a wide range of targets, syngeneic and allogeneic, of tumor and normal cell origin. Lysis was a cell-mediated phenomenon but was not blocked by anti-Ly-2. One H-2-negative target was lysed, but one was resistant. Xenogeneic (human) tumor cells were not lysed. The cells in the clones were large, vacuolated, granular lymphocytes. They originated from single Ly-2+ responder cells and not from irradiated filler cells. Therefore, activated lymphocyte killers and other natural killer-like cells may be differentiated elements of the Ly-2+ T-cell lineage.


Altered ligand rebinding kinetics due to distal-side effects in hemoglobin chico (Lysbeta66(E10) --> thr).
Hb Chico is an unusual human hemoglobin variant that has lowered oxygen affinity, but unaltered cooperativity and anion sensitivity. Previous studies showed these features to be associated with distal-side heme pocket alterations that confer increased structural rigidity on the molecule and that increase water content in the beta-chain heme pocket. We report here that the extent of nanosecond geminate rebinding of oxygen to the variant and its isolated beta-chains is appreciably decreased. Structural alterations in this variant decrease its oxygen recombination rates without significantly altering rates of migration out of the heme pocket. Data analysis indicates that one or more barriers that impede rebinding of oxygen from docking sites in the heme pocket are increased, with less consequence for CO rebinding. Resonance Raman spectra show no significant alterations in spectral regions sensitive to interactions between the heme iron and the proximal histidine residue, confirming that the functional differences in the variant are due to distal-side heme pocket alterations. These effects are discussed in the context of a schematic representation of heme pocket wells and barriers that could aid the design of novel hemoglobins with altered ligand affinity without loss of the normal allosteric responses that facilitate unloading of oxygen to respiring tissues.


Effects of smoking and smoking cessation on human serum metabolite profile: results from the KORA cohort study.
Metabolomics helps to identify links between environmental exposures and intermediate biomarkers of disturbed pathways. We previously reported variations in phosphatidylcholines in male smokers compared with non-smokers in a cross-sectional pilot study with a small sample size, but knowledge of the reversibility of smoking effects on metabolite profiles is limited. Here, we extend our metabolomics study with a large prospective study including female smokers and quitters. Using targeted metabolomics approach, we quantified 140 metabolite concentrations for 1,241 fasting serum samples in the population-based Cooperative Health Research in the Region of Augsburg (KORA) human cohort at two time points: baseline survey conducted between 1999 and 2001 and follow-up after seven years. Metabolite profiles were compared among groups of current smokers, former smokers and never smokers, and were further assessed for their reversibility after smoking cessation. Changes in metabolite concentrations from baseline to the follow-up were investigated in a longitudinal analysis comparing current smokers, never smokers and smoking quitters, who were current smokers at baseline but former smokers by the time of follow-up. In addition, we constructed protein-metabolite networks with smoking-related genes and metabolites. We identified 21 smoking-related metabolites in the baseline investigation (18 in men and six in women, with three overlaps) enriched in amino acid and lipid pathways, which were significantly different between current smokers and never smokers. Moreover, 19 out of the 21 metabolites were found to be reversible in former smokers. In the follow-up study, 13 reversible metabolites in men were measured, of which 10 were confirmed to be reversible in male quitters. Protein-metabolite networks are proposed to explain the consistent reversibility of smoking effects on metabolites. We showed that smoking-related changes in human serum metabolites are reversible after smoking cessation, consistent with the known cardiovascular risk reduction. The metabolites identified may serve as potential biomarkers to evaluate the status of smoking cessation and characterize smoking-related diseases.


The adaptor protein beta-arrestin2 enhances endocytosis of the low density lipoprotein receptor.
Endocytosis of the low density lipoprotein (LDL) receptor (LDLR) in coated pits employs the clathrin adaptor protein ARH. Similarly, agonist-dependent endocytosis of heptahelical receptors in coated pits employs the clathrin adaptor beta-arrestin proteins. In mice fed a high fat diet, we found that homozygous deficiency of beta-arrestin2 increased total and LDL plus intermediate-density lipoprotein cholesterol levels by 23 and 53%, respectively (p < 0.05), but had no effect on high density lipoprotein cholesterol levels. We therefore tested whether beta-arrestins could affect the constitutive endocytosis of the LDLR. When overexpressed in cells, beta-arrestin1 and beta-arrestin2 each associated with the LDLR, as judged by co-immunoprecipitation, and augmented LDLR endocytosis by approximately 70%, as judged by uptake of fluorescent LDL. However, physiologic expression levels of only beta-arrestin2, and not beta-arrestin1, enhanced endogenous LDLR endocytosis (by 65%) in stably transfected beta-arrestin1/beta-arrestin2 double-knockout mouse embryonic fibroblasts (MEFs). Concordantly, when RNA interference was used to suppress expression of beta-arrestin2, but not beta-arrestin1, LDLR endocytosis was reduced. Moreover, beta-arrestin2-/- MEFs demonstrated LDLR endocytosis that was 50% less than cognate wild type MEFs. In fusion protein pull-down assays, beta-arrestin2 bound to the LDLR cytoplasmic tail stoichiometrically, and binding was abolished by mutation of LDLR Tyr807 to Ala. Mutation of LDLR cytoplasmic tail Ser833 to Asp enhanced both the affinity of LDLR fusion protein binding to beta-arrestin2, and the efficiency of LDLR endocytosis in cells expressing beta-arrestin2 physiologically. We conclude that beta-arrestin2 can bind to and enhance endocytosis of the LDLR, both in vitro and in vivo, and may thereby influence lipoprotein metabolism.


Automatic assignment of metal-containing peptides in proteomic LC-MS and MS/MS data sets.
Transition metal-containing proteins and enzymes are critical for the maintenance of cellular function and metal-based (metallo)drugs are commonly used for the treatment of many diseases, such as cancer. Detection and characterisation of metallodrug targets is crucial for improving drug-design and therapeutic efficacy. Due to the unique isotopic ratios of many metal species, and the complexity of proteomic samples, standard MS data analysis of these species is unsuitable for accurate assignment. Herein a new method for differentiating metal-containing species within complex LCMS data is presented based upon the Smart Numerical Annotation Procedure (SNAP). SNAP-LC accounts for the change in isotopic envelopes for analytes containing non-standard species, such as metals, and will accurately identify, record, and display the particular spectra within extended LCMS runs that contain target species, and produce accurate lists of matched peaks, greatly assisting the identification and assignment of modified species and tailored to the metals of interest. Analysis of metallated species obtained from tryptic digests of common blood proteins after reactions with three candidate metallodrugs is presented as proof-of-concept examples and demonstrates the effectiveness of SNAP-LC for the fast and accurate elucidation of metallodrug targets.


Fluorogen-Activating-Protein-Loaded Tantalum Oxide Nanoshells for in Vivo On-Demand Fluorescence/Photoacoustic Imaging.
Optical imaging of targeted compartments within living animals has been widely adopted in many research areas. In particular, various fluorescence-based probes and emerged photoacoustic molecules that enable sensitive and specific imaging through tissue have greatly advanced clinically relevant studies. However, delivery and signal penetration have placed requirements on the performance of conventional optical probes. Here, we use hallow tantalum oxide (TaOx) nanoparticles to enclose fluorogen-activating protein (FAP) for the in vivo fluorescence and photoacoustic imaging of cancer cells. We found that the TaOx shell can provide a natural cover for the enclosed fluorogen/FAP complexes, protecting them from photobleaching and common biodegradation. Moreover, we have developed a near-infrared excitable tetrafluorinated photoacoustic fluorogen for the specific and persistent photoacoustic imaging of tumors. We believe that this enclosing and delivery strategy of optical biomolecules will be an attractive alternative for bioimaging.


Anti-science--a misunderstanding.
The modern revolt against science has been spurred by a misunderstanding of the nature of science, argues J. A. Passmore, professor of philosophy at the Australian National University, Canberra. Passmore believes that some of the misunderstanding rests with critics of science such as Theodore Roszak (see Science, 1 December 1972) and somne with the way that scientists have presented their affairs to the public. In the excerpts below (taken from the December issue of Search, the journal of the Australian Association for the Advancement of Science), Passmore analyzes the revolt against science as one that "condemns science for making itself the instrument of power, looks with dismay on the devastation to which science-based technology has given rise, rejects a world made gray by standardization . . . [and] seeks to reinstate the imagination and direct sensual enjoyment."


Suppression of inflammation by recombinant Salmonella typhimurium harboring CCL22 microRNA.
Atopic dermatitis (AD) is an inflammatory, chronically relapsing, puritic skin disorder. These syndromes result from multifactorial inheritance, with interaction between genetic and environmental factors. In particular, the macrophage-derived chemokine CCL22 is directly implicated in skin inflammatory reactions and its levels are significantly elevated in serum and correlated with disease severity in AD. We tested the suppression of the CCL22 gene by microRNA (miRNA) and observed the effects in mice with inflammation similar to AD. We used Salmonella as a vector to deliver miRNA. The recombinant strain of Salmonella typhimurium expressing CCL22 miRNA (ST-miRCCL22) was prepared for in vivo knockdown of CCL22. ST-miRCCL22 was orally inoculated into mice and the CCL22 gene suppressed with CCL22 miRNA in the activated lymphocytes. IgE and interleukin-4 were inhibited and interferon-γ was induced after treatments with ST-miRCCL22 and CCL22 was suppressed. Further, Th17 cells were suppressed in the atopic mice treated with ST-miRCCL22. These results suggested that suppression of the CCL22 gene using Salmonella induced anti-inflammatory effects.


Validity and inter-rater reliability of the Lindop Parkinson's Disease Mobility Assessment: a preliminary study.
To assess the validity and inter-rater reliability of the Lindop Parkinson's Disease Mobility Assessment (LPA); a scale developed to gather objective information on gait and bed mobility in patients with Parkinson's disease. Two therapists scored a group of patients with Parkinson's disease using the motor examination section of the Unified Parkinson's Disease Rating Scale (UPDRS-ME) and the LPA scale. The association between scores obtained using the two scales was calculated, and the scores obtained by the therapists using each scale were compared. A Parkinson's disease clinic in a day hospital in a district general hospital. Forty-nine subjects with Parkinson's disease were recruited (33 males, 16 females, mean age 75.84+/-7.16 years). There was a significant association between the LPA and UPDRS-ME data for both raters (Rater A, rho -0.67; Rater B, rho -0.63; P<0.001). The limits of agreement showed that the two raters scored within two scale points of each other on 95% of occasions, and that there was no systematic bias between raters. Percentage agreement between raters ranged from 82% to 100% for the LPA. The LPA took significantly less time to complete [mean 7.7 (standard deviation 2.9) minutes] than the UPDRS-ME [mean 10.5 (standard deviation 2.2) minutes; P<0.001]. The results suggest that the LPA is a valid measure with good inter-rater reliability. It is quick and easy to administer, and provides objective information about the gait and bed mobility of elderly patients with Parkinson's disease.


Association of Childhood Oral Infections With Cardiovascular Risk Factors and Subclinical Atherosclerosis in Adulthood.
Severe forms of common chronic oral infections or inflammations are associated with increased cardiovascular risk in adults. To date, the role of childhood oral infections in cardiovascular risk is not known because no long-term studies have been conducted. To investigate whether signs of oral infections in childhood are associated with cardiovascular risk factors and subclinical atherosclerosis in adulthood. The cohort study (n = 755) was derived from the Cardiovascular Risk in Young Finns Study, an ongoing prospective cohort study in Finland initiated in 1980. Participants underwent clinical oral examinations during childhood, when they were aged 6, 9, or 12 years and a clinical cardiovascular follow-up in adulthood in 2001 at age 27, 30, or 33 years and/or in 2007 at age 33, 36, or 39 years. Cardiovascular risk factors were measured at baseline and during the follow-up until the end of 2007. Final statistical analyses were completed on February 19, 2019. Four signs of oral infections (bleeding on probing, periodontal probing pocket depth, caries, and dental fillings) were documented. Cumulative lifetime exposure to 6 cardiovascular risk factors was calculated from dichotomized variables obtained by using the area-under-the-curve method. Subclinical atherosclerosis (ie, carotid artery intima-media thickness [IMT]) was quantified in 2001 (n = 468) and 2007 (n = 489). This study included 755 participants, of whom 371 (49.1%) were male; the mean (SD) age at baseline examination was 8.07 (2.00) years. In this cohort, 33 children (4.5%) had no sign of oral infections, whereas 41 (5.6%) had 1 sign, 127 (17.4%) had 2 signs, 278 (38.3%) had 3 signs, and 248 (34.1%) had 4 signs. The cumulative exposure to risk factors increased with the increasing number of oral infections both in childhood and adulthood. In multiple linear regression models, childhood oral infections, including signs of either periodontal disease (R2 = 0.018; P = .01), caries (R2 = 0.022; P = .008), or both (R2 = 0.024; P = .004), were associated with adulthood IMT. The presence of any sign of oral infection in childhood was associated with increased IMT (third tertile vs tertiles 1 and 2) with a relative risk of 1.87 (95% CI, 1.25-2.79), whereas the presence of all 4 signs produced a relative risk of 1.95 (95% CI, 1.28-3.00). The associations were more obvious in boys: if periodontal disease were present, the corresponding estimate was 1.69 (95% CI, 1.21-2.36); if caries, 1.46 (95% CI, 1.04-2.05); and if all 4 signs of oral infections, 2.25 (95% CI, 1.30-3.89). The associations were independent of cardiovascular risk factors. Oral infections in childhood appear to be associated with the subclinical carotid atherosclerosis seen in adulthood.


The diagnostic role of procalcitonin and other biomarkers in discriminating infectious from non-infectious fever.
Fever is not only observed in the course of a bacterial or viral infection, but can be a symptom of, for instance, auto-immune, malignant or thromboembolic disease. Determining the etiology of fever in a fast and reliable way is of pivotal importance, as different causes of fever may ask for different therapies. Neither clinical signs and symptoms, nor traditional biomarkers, such as CRP, leukocytes and ESR have sufficient sensitivity and specificity to guide treatment decisions. In this review we focus on the value of traditional and newer biomarkers in non-infectious febrile diseases. Procalcitonin (PCT) seems to be the most helpful laboratory marker for the differentiation of causes of fever, particularly in autoimmune, autoinflammatory and malignant diseases.


Impact of Predialysis Psychosocial Conditions on Kidney Transplant Recipient Survival: Evidence Using Propensity Score Matching.
An increased number of end-stage renal disease patients suffer psychosocial conditions and may experience delayed access to transplantation due to listing restrictions. However, it remains to be shown whether preexisting psychosocial conditions confer an independent risk factor of poor posttransplant outcomes. We addressed this gap in knowledge by conducting a retrospective cohort study to investigate an independent association between the risk of death after transplant and having a diagnosis of psychosocial conditions 1 year prior to starting dialysis. All cases of adult deceased-donor kidney transplantation performed in Ontario, Canada, between April 1, 2002, and March 31, 2013, were used. Propensity score matching was applied to adjust for potential endogenous bias of using predialysis psychosocial status to predict posttransplant mortality. Survival analysis techniques, including Kaplan-Meier curves and Cox proportional hazards modeling, were also used. Our results indicate a 49.4% (hazard ratio [HR] = 1.494 [95% confidence interval (CI) = 1.168-1.913]) increased relative risk of posttransplant death to be associated with predialysis psychosocial conditions, when other factors are held constant. The effect is significant (P = .001) and is independent of other known predictors of death including advanced age. Findings from this study offered strong support for the development of psychosocial evaluation to screen candidates prior to transplant listing and early interventions for transplant candidates with psychosocial concerns.


Sprouting of corneal sensory fibers in rats treated at birth with capsaicin.
Normal structure and function of corneal epithelium is known to be related to proper innervation. To investigate possible trophic actions of sensory neurons on corneal epithelium, corneal innervation and various physical parameters were studied in normal rats and in rats treated as neonates with intraperitoneal injections of capsaicin. Corneal lesions were noted in treated rats which varied from multiple punctate areas of corneal opacity to deep stromal opacity with ulceration and neovascularization. These lesions waxed and waned throughout the animal's life. In addition, mechanical threshold of the corneal reflex was higher in capsaicin-treated rats. The tear rate in response to a provocative test was diminished in treated rats, presumably due to reduced afferent trigeminal input to the brain stem; blinking rates were more frequent in these animals. Using fluorescent retrograde tracing techniques, the number of cells innervating the cornea in capsaicin-treated rats was found to be significantly less compared with control animals. Innervation in the cornea (examined using a gold chloride technique) demonstrated a decrease in the number of corneal large axons in treated rats with neurite sprouting of these axons yielding a higher density of nerve fibers compared with controls. Thus, sprouting of residual sensory neurons occurs in response to the partial corneal denervation produced by capsaicin, and this sprouting does not functionally compensate to prevent the development of chronic keratitis.


c-Myc is a novel target of cell cycle arrest by honokiol in prostate cancer cells.
Honokiol (HNK), a highly promising phytochemical derived from Magnolia officinalis plant, exhibits in vitro and in vivo anticancer activity against prostate cancer but the underlying mechanism is not fully clear. This study was undertaken to delineate the role of c-Myc in anticancer effects of HNK. Exposure of prostate cancer cells to plasma achievable doses of HNK resulted in a marked decrease in levels of total and/or phosphorylated c-Myc protein as well as its mRNA expression. We also observed suppression of c-Myc protein in PC-3 xenografts upon oral HNK administration. Stable overexpression of c-Myc in PC-3 and 22Rv1 cells conferred significant protection against HNK-mediated growth inhibition and G0-G1 phase cell cycle arrest. HNK treatment decreased expression of c-Myc downstream targets including Cyclin D1 and Enhancer of Zeste Homolog 2 (EZH2), and these effects were partially restored upon c-Myc overexpression. In addition, PC-3 and DU145 cells with stable knockdown of EZH2 were relatively more sensitive to growth inhibition by HNK compared with control cells. Finally, androgen receptor overexpression abrogated HNK-mediated downregulation of c-Myc and its targets particularly EZH2. The present study indicates that c-Myc, which is often overexpressed in early and late stages of human prostate cancer, is a novel target of prostate cancer growth inhibition by HNK.


Using DNA fingerprints to infer familial relationships within NHANES III households.
Developing, targeting, and evaluating genomic strategies for population-based disease prevention require population-based data. In response to this urgent need, genotyping has been conducted within the Third National Health and Nutrition Examination (NHANES III), the nationally-representative household-interview health survey in the U.S. However, before these genetic analyses can occur, family relationships within households must be accurately ascertained. Unfortunately, reported family relationships within NHANES III households based on questionnaire data are incomplete and inconclusive with regards to actual biological relatedness of family members. We inferred family relationships within households using DNA fingerprints (Identifiler(R)) that contain the DNA loci used by law enforcement agencies for forensic identification of individuals. However, performance of these loci for relationship inference is not well understood. We evaluated two competing statistical methods for relationship inference on pairs of household members: an exact likelihood ratio relying on allele frequencies to an Identical By State (IBS) likelihood ratio that only requires matching alleles. We modified these methods to account for genotyping errors and population substructure. The two methods usually agree on the rankings of the most likely relationships. However, the IBS method underestimates the likelihood ratio by not accounting for the informativeness of matching rare alleles. The likelihood ratio is sensitive to estimates of population substructure, and parent-child relationships are sensitive to the specified genotyping error rate. These loci were unable to distinguish second-degree relationships and cousins from being unrelated. The genetic data is also useful for verifying reported relationships and identifying data quality issues. An important by-product is the first explicitly nationally-representative estimates of allele frequencies at these ubiquitous forensic loci.


Hepatic transcatheter arterial chemoembolization alternating with systemic protracted continuous infusion 5-fluorouracil for gastrointestinal malignancies metastatic to liver: a phase II trial of the Puget Sound Oncology Consortium (PSOC 1104).
We assessed a regimen of alternating regional and systemic therapy in patients with gastrointestinal malignancies with liver-dominant metastases for feasibility, toxicity, response rate, response duration, patterns of progression, and progression-free and overall survival. Regional therapy comprised selective hepatic transcatheter arterial chemoembolization (TACE) using a suspension of cisplatin and particulate polyvinyl alcohol. This procedure was delivered between cycles of protracted continuous infusion 5-fluorouracil (PCI-5FU) as systemic chemotherapy. Patient eligibility criteria included: (a) having histologically documented adenocarcinoma arising from a gastrointestinal primary site with unresectable liver metastases bidimensionally measurable on computerized tomography scan; (b) age greater than 18 years; and (c) performance status 0-2 (Zubrod). PCI-5FU (250 mg/m2/day) was administered i.v. for 28 days, followed by the first TACE (TACE 1) delivered to the hepatic artery supplying the lobe with the greatest tumor burden. Restaging was performed before TACE 2 and TACE 3, which followed at monthly intervals. PCI-5FU for 21 days was sandwiched between each of the TACE treatments. After the final TACE, maintenance PCI-5FU was given for 28 days of each 35-day cycle until toxicity or progression. Between December 23, 1991, and January 19, 1995, 32 patients were registered in this trial, of whom 27 were eligible; 20 completed one or more treatment cycles and were evaluable for radiographic response. Patients with colorectal liver metastases predominated (74%). Twelve (44%) of 27 patients had failed one or more prior treatment regimens. There were no treatment-related deaths, and hematological and hepatic toxicities were generally manageable and reversible. Two patients, however, developed hepatic abscesses requiring drainage, and one patient developed an infarcted gallbladder, which necessitated cholecystectomy. There were no patients with complete responses; there were 8 (40%) with partial responses, 4 (20%) with minor responses, 2 (10%) with stable disease, and 6 (30%) who progressed on the treatment. The median duration of response for partial responders was 4.2 months (127 days; range, 56-245 days). The median reduction in carcinoembryonic antigen for responders was 87.5%. Two patients underwent subsequent resection of residual metastases; one of them is still alive at 58.4 months follow-up. The predominant site of disease progression was the liver; 25% of the patients progressed in extrahepatic sites. The median overall survival for the whole group is 14.3 months (95% confidence interval, 7.2-16.2). Actuarial overall survival for the whole group at 1 year and 2 years is 57 and 19%, respectively. Alternating systemic PCI-5FU and regional TACE (cisplatin/polyvinyl alcohol) is an active and feasible regimen with manageable toxicities in patients with metastatic gastrointestinal malignancies with liver-dominant disease and merits further investigation. The complications seen were in line with those reported at other specialized centers.


An Integrative Genomic Analysis of Formalin Fixed Paraffin-Embedded Archived Serous Ovarian Carcinoma Comparing Long-term and Short-term Survivors.
This study aimed to perform an integrative genetic analysis of patients with matched serous ovarian cancer having long-term or short-term survival using formalin fixed paraffin-embedded (FFPE) tissue samples. All patients with serous ovarian carcinoma who underwent surgery between 1998 and 2007 at the Department of Gynaecology, Uppsala University Hospital, Sweden were considered. From this cohort, we selected biomaterial from 2 groups of patients with long-term and short-term survival matched for age, stage, histologic grade, and outcome of surgery. Genomic DNA from FFPE sample was analyzed with SNP array and targeted next-generation sequencing of 26 genes. Forty-three samples (primary tumors and metastases) from 23 patients were selected for genomic profiling, the survival in the subgroups were 134 and 36 months, respectively. We observed a tendency toward increased genomic instability in those with long-term survival with higher proportion of somatic copy number alterations (P = 0.083) and higher average ploidy (P = 0.037). TP53 mutations were found in 50% of the patients. Frequency of TP53 mutations did not differ between the survival groups (P = 0.629). We validated both previous genomic findings in ovarian cancer and the proposed association between increased genomic instability and better survival. These results exemplify that analysis of genomic biomarkers is feasible on archived FFPE tissue.


Development of the COVID-19 Real-Time Information System for Preparedness and Epidemic Response (CRISPER), Australia.
Accurate and current information has been highlighted across the globe as a critical requirement for the COVID-19 pandemic response. To address this need, many interactive dashboards providing a range of different information about COVID-19 have been developed. A similar tool in Australia containing current information about COVID-19 could assist general practitioners and public health responders in their pandemic response efforts. The COVID-19 Real-time Information System for Preparedness and Epidemic Response (CRISPER) has been developed to provide accurate and spatially explicit real-time information for COVID-19 cases, deaths, testing and contact tracing locations in Australia. Developed based on feedback from key users and stakeholders, the system comprises three main components: (1) a data engine; (2) data visualization and interactive mapping tools; and (3) an automated alert system. This system provides integrated data from multiple sources in one platform which optimizes information sharing with public health responders, primary health care practitioners and the general public.


Social skills and problem-solving training for children with early-onset conduct problems: who benefits?
Families of 99 children with early-onset conduct problems, aged 4-8 years, were randomly assigned to a child training treatment group (CT) utilizing the Incredible Years Dinosaur Social Skills and Problem Solving Curriculum or a waiting-list control group (CON). Post-treatment CT children had significantly fewer externalizing problems at home, less aggression at school, more prosocial behavior with peers, and more positive conflict management strategies than CON children. Significantly more CT than CON children showed clinically significantly improvements on reports and independent observations of aggressive and noncompliant behavior. The differential treatment response was evaluated according to child comorbidity with attention deficit hyperactivity disorder (ADHD), parenting discipline practices. and family risk factors. The only risk factor related to failure to make improvements in child conduct problems after treatment was negative parenting (i.e., maternal critical statements and physical force). The long-term follow-up 1 year later indicated that most of the significant post-treatment changes were maintained.


Role of Omega-3 Fatty Acids on Lipid Profile in Diabetic Dyslipidaemia: Single Blind, Randomised Clinical Trial.
Diabetic dyslipidaemia is characterised by hypertriglyceridaemia, low High Density Lipoprotein (HDL), postprandial lipimea, small and dense LDL particles is considered to be a major predisposing factor for various macrovascular complications. Omega-3 fatty acids are fish oil derivative introduced in the market for dyslipidaemia associated with increased triglyceride level. To study the effect of omega-3 fatty acids on lipid profile in Type II diabetes patients. This study was prospective, single blind, randomized comparative trial. Hundred patients were randomized into three groups. Group I received metformin 500 mg twice daily and placebo, Group II received metformin 500 mg twice daily and omega-3 fatty acids (1 gram) once daily and the Group III received metformin 500 mg twice daily and omega-3 fatty acids (1 gram) twice daily. ANOVA test was applied for analysis. Group II was effective in reducing the triglyceride level from 144.59±14.18 mg/dl to 101±13.31 mg/dl which was significant as compared to Group I from 147.67±18.57 mg/dl to 145.8±19.86 mg/dl respectively. Group III containing 1 g of omega-3 fatty acids twice daily showed decrease from 144.83±22.17 mg/dl to 86±17.46 mg/dl and was more effective in reducing triglyceride levels than Group II containing 1 gram of omega-3 fatty acids once daily. Omega-3 fatty acids can be given in conjunction with metformin to reduce triglyceride levels in diabetic dyslipidaemia without any adverse drug reactions or any drug interaction. Omega-3 fatty acids were effective in reducing the triglyceride level significantly as compared to placebo. Two grams of omega-3 fatty acids were more effective than 1 gram of omega-3 fatty acids in reducing triglyceride levels.


Experimental evaluation of staple lines in gastric surgery.
This study was designed to evaluate the bursting pressure, tensile strength, and healing of staple lines in the dog stomach. Pavlov pouches were created in 30 dogs using a single, double, or triple application of a stapler loaded with 4.8-mm staples. Staple lines were assessed immediately in half of the animals and after 21 days in the remaining animals. Acutely, reinforcing a single staple line with a second or third application of the stapler resulted in an anastomosis more resistant to leakage and disruption. Satisfactory healing occurred by 21 days and contributed to staple line strength in the pouches using a single and double application of the stapler. The orientation of collagen deposition in the anastomosis was related to the number of staples used. The degree of collagen deposition across the muscularis mucosa correlated with the healed staple line strength. Although healing occurred when a triple application of the stapler was utilized, the orientation of the fibroconnective tissue was parallel to the anastomosis and did not contribute to staple line integrity.


Girls on the Run: Impact of a Physical Activity Youth Development Program on Psychosocial and Behavioral Outcomes.
Girls on the Run is an after-school physical activity-based positive youth development program designed to enhance girls' social, psychological, and physical development. We evaluated the effectiveness of the program by employing a longitudinal design and mixed methods. Girls (N = 203; aged 8-11 y) completed survey measures of positive youth development constructs (competence, confidence, connection, character, and caring), physical activity, and sedentary behavior prior to, at the end of, and 3 months after the season. Subsamples of girls, coaches, caregivers, and school personnel participated in focus groups. Coaches completed information about their team's community impact project and number of girls who completed the season-ending 5k. The full sample improved in confidence and connection, whereas girls who started below the preseason average showed the greatest gains from preseason to postseason on all measures, and scores were maintained or continued to improve at follow-up. All stakeholders in focus groups corroborated evidence of season-long improvement in social and emotional behaviors and health outcomes. Involvement in the community impact project contributed to girls' growth in character and empathy skills. Findings provide empirical evidence that Girls on the Run is effective in promoting positive youth development, including season-long and lasting change in competence, confidence, connection, character, caring, and physical activity, especially among girls who exhibited lower preseason scores than their peers.


Performance of a biofilter system with agave fiber filter media for municipal wastewater treatment.
Agave plants grow in semi-arid regions and are used for mescal production. However, agave fiber by-products are considered waste materials. Thus, we tested agave fiber as a filter media and biofilm material carrier for removing pollutants from municipal wastewater. Three laboratory-scale biofiltration reactors were used in two trials with five hydraulic loading rates (HLRs = 0.27, 0.54, 0.80, 1.07 and 1.34 m(3) m(-2) d(-1)). One series was conducted using mechanical aeration (0.62 m(3) m(-2) h(-1)). To prevent compaction, decreasing pressure and clogging of the filter media, 4, 8 and 12 internal divisions were evaluated in the biofilter column. After 17 months of continuous operation at an HLR of 0.80 m(3) m(-2) d(-1), the removal efficiencies of the aerated biofilters were 92.0% biochemical oxygen demand, 79.7% chemical oxygen demand, 98.0% helminth eggs, 99.9% fecal coliforms and 91.9% total suspended solids. Statistical analysis showed that the chosen operational parameters significantly influenced the removal efficiencies of the biofilters. The effluent quality obtained under these conditions complied with the Mexican and US EPA standards for agricultural irrigation and green spaces, except for coliforms, which is why the effluents must be disinfected. Thus, agave fiber is a favorable choice for use as a packing material in biofiltration processes.


Identification of novel and zoonotic Cryptosporidium species in fish from Papua New Guinea.
There is still limited information on the distribution of Cryptosporidium species and genotypes in fish. The present study investigated the prevalence of Cryptosporidium species in cultured freshwater (n=132), wild freshwater (n=206) and wild marine (n=276) fish in Papua New Guinea (PNG) by PCR screening at the 18S rRNA locus. A total of seven fish (2 cultured freshwater, 1 wild freshwater and 4 wild marine fish) were identified as positive for Cryptosporidium. Specifically, Cryptosporidium was found in four different host species (Nile tilapia, Oreochromis niloticus; silver barb, Puntius gonionotus; mackerel scad, Decapterus maracellus and oblong silver biddy, Gerres oblongus), giving an overall prevalence of 1.14% (95% CI: 0.3-2%, n=7/614). Of the seven positive isolates, five were identified as C. parvum and two were a novel piscine genotype, which we have named piscine genotype 8. Piscine genotype 8 was identified in two marine oblong silver biddies and exhibited 4.3% genetic distance from piscine genotype 3 at the 18S locus. Further subtyping of C. parvum isolates at the 60 kDa glycoprotein (gp60) locus identified 3 C. parvum subtypes (IIaA14G2R1, IIaA15G2R1 and IIaA19G4R1) all of which are zoonotic and a C. hominis subtype (IdA15G1). The zoonotic Cryptosporidium were identified in fish samples from all three groups; cultured and wild freshwater and wild marine fish. Detection of Cryptosporidium among aquaculture fingerlings warrants further research to gain a better understanding of the epidemiology of Cryptosporidium infection in cultured fish. The identification of zoonotic Cryptosporidium genotypes in fish from PNG has important public health implications and should be investigated further.


Suppression of Proinflammatory and Prosurvival Biomarkers in Oral Cancer Patients Consuming a Black Raspberry Phytochemical-Rich Troche.
Black raspberries (BRB) demonstrate potent inhibition of aerodigestive tract carcinogenesis in animal models. However, translational clinical trials evaluating the ability of BRB phytochemicals to impact molecular biomarkers in the oral mucosa remain limited. The present phase 0 study addresses a fundamental question for oral cancer food-based prevention: Do BRB phytochemicals successfully reach the targeted oral tissues and reduce proinflammatory and antiapoptotic gene expression profiles? Patients with biopsy-confirmed oral squamous cell carcinomas (OSCC) administered oral troches containing freeze-dried BRB powder from the time of enrollment to the date of curative intent surgery (13.9 ± 1.27 days). Transcriptional biomarkers were evaluated in patient-matched OSCCs and noninvolved high at-risk mucosa (HARM) for BRB-associated changes. Significant expression differences between baseline OSCC and HARM tissues were confirmed using a panel of genes commonly deregulated during oral carcinogenesis. Following BRB troche administration, the expression of prosurvival genes (AURKA, BIRC5, EGFR) and proinflammatory genes (NFKB1, PTGS2) were significantly reduced. There were no BRB-associated grade 3-4 toxicities or adverse events, and 79.2% (N = 30) of patients successfully completed the study with high levels of compliance (97.2%). The BRB phytochemicals cyanidin-3-rutinoside and cyanidin-3-xylosylrutinoside were detected in all OSCC tissues analyzed, demonstrating that bioactive components were successfully reaching targeted OSCC tissues. We confirmed that hallmark antiapoptotic and proinflammatory molecular biomarkers were overexpressed in OSCCs and that their gene expression was significantly reduced following BRB troche administration. As these molecular biomarkers are fundamental to oral carcinogenesis and are modifiable, they may represent emerging biomarkers of molecular efficacy for BRB-mediated oral cancer chemoprevention.


A review of biosensing techniques for detection of trace carcinogen contamination in food products.
Carcinogen contaminations in the food chain, for example heavy metal ions, pesticides, acrylamide, and mycotoxins, have caused serious health problems. A major objective of food-safety research is the identification and prevention of exposure to these carcinogens, because of their impossible-to-reverse tumorigenic effects. However, carcinogen detection is difficult because of their trace-level presence in food. Thus, reliable and accurate separation and determination methods are essential to protect food safety and human health. This paper summarizes the state of the art in separation and determination methods for analyzing carcinogen contamination, especially the advances in biosensing methods. Furthermore, the application of promising technology including nanomaterials, imprinted polymers, and microdevices is detailed. Challenges and perspectives are also discussed.


Transfusion-free interval is associated with improved survival in patients with higher-risk myelodysplastic syndromes engaged in routine care.
Most higher-risk myelodysplastic syndrome (HR-MDS) patients will become transfusion-dependent, leading to potential complications, including infections or end-organ dysfunction. Data correlating achievement of transfusion-free intervals (TFIs) during first-line therapy (1LT) with survival are sparse. We evaluated HR-MDS patients receiving 1LT diagnosed from 1/1/2008 to 7/31/2015 and the impact of a TFI (≥60-day interval without transfusions) on progression-free and overall survival (PFS, OS) using Cox proportional-hazard models. Two hundred and twenty-nine HR-MDS patients received 1LT; overall, median PFS/OS were 8.4 months and 14.7 months, respectively. Two-year PFS/OS were 22.3% and 34.6%, respectively. Median PFS/OS were longer for patients with vs. without a TFI (16.9 vs. 6.1 months and 26.1 vs. 11.8 months, respectively; p < .01 [both]). Two-year PFS (43.0% vs. 3.9%; p < .01) and 2-year OS (51.8% vs. 22.5%; p < .01) were also longer in patients with a TFI vs. not. Achievement of a TFI during 1LT appears to positively affect PFS and OS in HR-MDS patients.


A possible central mechanism in autism spectrum disorders, part 2: immunoexcitotoxicity.
In this section, I explore the effects of mercury and inflammation on transsulfuration reactions, which can lead to elevations in androgens, and how this might relate to the male preponderance of autism spectrum disorders (ASD). It is known that mercury interferes with these biochemical reactions and that chronically elevated androgen levels also enhance the neurodevelopmental effects of excitotoxins. Both androgens and glutamate alter neuronal and glial calcium oscillations, which are known to regulate cell migration, maturation, and final brain cytoarchitectural structure. Studies have also shown high levels of DHEA and low levels of DHEA-S in ASD, which can result from both mercury toxicity and chronic inflammation. Chronic microglial activation appears to be a hallmark of ASD. Peripheral immune stimulation, mercury, and elevated levels of androgens can all stimulate microglial activation. Linked to both transsulfuration problems and chronic mercury toxicity are elevations in homocysteine levels in ASD patients. Homocysteine and especially its metabolic products are powerful excitotoxins. Intimately linked to elevations in DHEA, excitotoxicity and mercury toxicity are abnormalities in mitochondrial function. A number of studies have shown that reduced energy production by mitochondria greatly enhances excitotoxicity. Finally, I discuss the effects of chronic inflammation and elevated mercury levels on glutathione and metallothionein.


Microbial electrodialysis cell for simultaneous water desalination and hydrogen gas production.
A new approach to water desalination is to use exoelectrogenic bacteria to generate electrical power from the biodegradation of organic matter, moving charged ions from a middle chamber between two membranes in a type of microbial fuel cell called a microbial desalination cell. Desalination efficiency using this approach is limited by the voltage produced by the bacteria. Here we examine an alternative strategy based on boosting the voltage produced by the bacteria to achieve hydrogen gas evolution from the cathode using a three-chambered system we refer to as a microbial electrodialysis cell (MEDC). We examined the use of the MEDC process using two different initial NaCl concentrations of 5 g/L and 20 g/L. Conductivity in the desalination chamber was reduced by up to 68 ± 3% in a single fed-batch cycle, with electrical energy efficiencies reaching 231 ± 59%, and maximum hydrogen production rates of 0.16 ± 0.05 m(3) H(2)/m(3) d obtained at an applied voltage of 0.55 V. The advantage of this system compared to a microbial fuel cell approach is that the potentials between the electrodes can be better controlled, and the hydrogen gas that is produced can be used to recover energy to make the desalination process self-sustaining with respect to electrical power requirements.


United Against Rabies Forum: The One Health Concept at Work.
Human deaths from rabies are preventable and can be eliminated by applying a systematic One Health approach. However, this ancient disease still threatens the lives of millions of people in up to 150 countries and kills an estimated 59, 000 people every year. Rabies today is largely a disease of poverty, almost always linked to dog bites, with most deaths occurring in neglected communities in Africa and Asia. The disease places an immense economic burden on its victims, a cost that far outweighs the investment needed to control it. A global framework for rabies elimination in humans is set out in Zero by 30: The Global Strategic Plan to end human deaths from dog-mediated rabies by 2030. Despite the existence of proven control strategies and agreement on the path to eliminating human rabies deaths, mortality numbers from rabies remain high, and COVID-19 has set back efforts even further. But COVID-19 has also highlighted the value of a One Health approach to zoonotic disease and pandemic prevention. Rabies control programs offer a practical route to building One Health capacities that can also address other zoonotic threats, including those with pandemic potential. The United Against Rabies Forum aims to accelerate progress on rabies elimination while applying a One Health approach. The Forum promotes cross-sector collaboration among stakeholders and supports countries in their rabies elimination efforts. Increased political engagement and resource mobilization, both internationally and nationally, will be needed to achieve global rabies goals and can also make One Health implementation a reality.


Membrane-bound carbonic anhydrase in human retinal pigment epithelium.
Inhibition of carbonic anhydrase (CA) by acetazolamide causes a decrease in the standing potential of the retinal pigment epithelium (RPE) and an increase in the rate of subretinal fluid absorption, and it may improve cystoid macular edema. These effects are thought to be mediated by the RPE. Given the solubility coefficient of acetazolamide, the drug is most likely to act by direct inhibition of membrane-bound CA (CA IV). To identify a substrate for acetazolamide in the RPE, the distribution of CA activity and the isoform of CA in the RPE membrane were investigated. Carbonic anhydrase activity was determined by Hansson's technique in fresh human eyes from donors of both sexes and different ages. The presence of the membrane-bound isoform CA IV was investigated immunohistochemically at the light and electron microscopic level, as well as by Western blotting in fresh RPE, and in adult and fetal RPE cultures. Hansson's histochemical method demonstrated CA activity on the apical and basolateral cell membrane of the RPE. Using the gamma-globulin fraction of a polyclonal antibody against pure CA IV, immunocytochemistry showed labeling for CA IV on the apical RPE membrane or morphologically polarized human adult and fetal RPE cultures. Gel electrophoresis and Western blotting demonstrated a major immunoreactive band at 55 kDa in homogenates, which was consistently reduced to approximately 35 kDa by incorporation of 0.1% Triton X-100 detergent. These results suggest that the clinical effects of carbonic anhydrase inhibitors on RPE function may be mediated via membrane-bound carbonic anhydrase activity in RPE and that CA IV is responsible for activity on the apical surface.


Herpesvirus tegument protein activates NF-kappaB signaling through the TRAF6 adaptor protein.
Viruses remodel the host cell to optimize their replication both by delivery of virion proteins into the cell and by de novo expression of viral proteins. The HSV particle contains several proteins that function to prepare the host cell for viral replication, including the VP16 transcriptional activator protein and virion host shutoff protein. HSV infection activates NF-kappaB pathways through Toll-like receptor (TLR) 2 and non-TLR pathways, and NF-kappaB activity is required for efficient viral replication. In a screen of the HSV proteome, we observed that the HSV U(L)37 tegument protein activates NF-kappaB signaling in a TLR2-independent manner. Expression of U(L)37 in transfected cells leads to IkappaB degradation and activation of both reporter genes and the endogenous IL-8 gene. This activation requires TNF receptor-associated factor 6 (TRAF6), and U(L)37 contains a TRAF6-binding domain that is required for interaction with TRAF6 and activation of NF-kappaB. A mutant virus encoding U(L)37 with an altered TRAF6-binding site shows reduced NF-kappaB activation in the early phase of infection. Therefore, the HSV U(L)37 virion structural protein can activate NF-kappaB through TRAF6. Activation of NF-kappaB by a virion tegument protein that is delivered into the host cell cytoplasm during viral entry represents a mechanism for activation of this pathway by a virus.


Influence of Music on Anxiety Induced by Fear of Heights in Virtual Reality.
Music is a potent mood regulator that can induce relaxation and reduce anxiety in different situations. While several studies demonstrate that certain types of music have a subjective anxiolytic effect, the reported results from physiological responses are less conclusive. Virtual reality allows us to study diverse scenarios of real life under strict experimental control while preserving high ecological validity. We aimed to study the modulating effect of music on the anxiety responses triggered by an immersive virtual reality scenario designed to induce fear of heights. Subjects experienced a virtual scenario depicting an exterior elevator platform ascending and descending the total height of its 350 meters tall supporting structure. Participants were allocated to either a group that experienced the elevator ride with background music or without, in a between-groups design. Furthermore, each group included participants with different degrees of fear of heights, ranging from low to high fear. Recordings of heart rate, galvanic skin response, body balance, and head movements were obtained during the experiments. Subjective anxiety was measured by means of three questionnaires. The scenario produced significant changes in subjective and physiological measures, confirming its efficacy as a stressor. A significant increase in state anxiety was found between pre and post-assessment in the silence group, but not in the music group, indicating that post-stress recovery was faster in the musical group. Results suggest that music can ameliorate the subjective anxiety produced by fear of heights.


Transcriptional and posttranscriptional activation of urokinase plasminogen activator gene expression in metastatic tumor cells.
Urokinase plasminogen activator (uPA) is a serine protease which has frequently been implicated in the process of tumor cell invasion and metastasis. The degree of expression and mode(s) of regulation of the uPA gene in metastatic compared with nonmetastatic tumor cells have not yet been addressed. We have cloned and sequenced a full-length rat uPA complementary DNA and utilized Northern blot analysis to report that the uPA gene is expressed at levels 3.5- to 70-fold higher in metastatic cell lines than in nonmetastatic cell lines derived from two independent rat mammary adenocarcinomas. Nuclear run-on assays and RNA half-life estimations indicated that metastatic MAT 13762 rat mammary adenocarcinoma cells expressed 3.5-fold higher levels of uPA RNA than a nonmetastatic derivative (J-clone), due to a combined increase in uPA gene transcription and cytoplasmic RNA stability. By contrast, uPA RNA (and enzyme) levels were elevated by up to 70-fold in metastatic clones of dimethylbenz(a)anthracene-induced rat mammary adenocarcinoma (DMBA-8) due to predominantly posttranscriptional mechanisms. Moreover, treatment of nonmetastatic DMBA-8 cell lines with protein synthesis inhibitors led to an increase in nuclear and cytoplasmic uPA RNA levels, without altering the rate of uPA gene transcription. These results suggest that in addition to gene transcription, posttranscriptional events localized in the nucleus and cytoplasm are key determinants of uPA gene activation in rat mammary adenocarcinomas.


Increase in susceptibility to insecticides with aging of wild Anopheles gambiae mosquitoes from Côte d'Ivoire.
Appropriate monitoring of vector insecticide susceptibility is required to provide the rationale for optimal insecticide selection in vector control programs. In order to assess the influence of mosquito age on susceptibility to various insecticides, field-collected larvae of An. gambiae s.l. from Tiassalé were reared to adults. Females aged 1, 2, 3, 5 and 10 days were exposed to 5 insecticides (deltamethrin, permethrin, DDT, malathion and propoxur) using WHO susceptibility test kits. Outcome measures included the LT50 (exposure time required to achieve 50% knockdown), the RR (resistance ratio, i.e. a calculation of how much more resistant the wild population is compared with a standard susceptible strain) and the mortality rate following 1 hour exposure, for each insecticide and each mosquito age group. There was a positive correlation between the rate of knockdown and mortality for all the age groups and for all insecticides tested. For deltamethrin, the RR50 was highest for 2 day old and lowest for 10 day old individuals. Overall, mortality was lowest for 2 and 3 day old individuals and significantly higher for 10 day old individuals (P < 0.05). With permethrin, the RR50 was highest for 1 to 3 day old individuals and lowest for 10 day old individuals and mortality was lowest for 1 to 3 day old individuals, intermediate for 5 day old and highest for 10 day old individuals. DDT did not display any knockdown effect and mortality was low for all mosquito age groups (<7%). With malathion, the RR50 was low (1.54 - 2.77) and mortality was high (>93%) for all age groups. With propoxur, no knockdown effect was observed for 1, 2 and 3 day old individuals and a very low level of mortality was observed (< 4%), which was significantly higher for 5 and 10 day old individuals (30%, P < 0.01). Results indicate that for An. gambiae s.l. adults derived from wild-collected larvae, there was an influence of age on insecticide susceptibility status, with younger individuals (1 to 3 days old) more resistant than older mosquitoes. This indicates that the use of 1 - 2 day old mosquitoes in susceptibility assays as recommended by the WHO should facilitate detection of resistance at the stage where the highest rate of the resistance phenotype is present.


How aging and bilingualism influence language processing: theoretical and neural models.
Healthy non-pathological aging is characterized by cognitive and neural decline, and although language is one of the more stable areas of cognition, older adults often show deficits in language production, showing word finding failures, increased slips of the tongue, and increased pauses in speech. Overall, research on language comprehension in older healthy adults show that it is more preserved than language production. Bilingualism has been shown to confer a great deal of neuroplasticity across the life span, including a number of cognitive benefits especially in executive functions such as cognitive control. Many models of bilingual language processing have been proposed to explain bilingual language processing. However, the question remains open of how such models might be modulated by age-related changes in language. Here, we discuss how current models of language processing in non-pathological aging, and models of bilingual language processing can be integrated to provide new research directions.


Challenges of using nationally representative, population-based surveys to assess rural cancer disparities.
Population-based surveys provide important information about cancer-related health behaviors across the cancer care continuum, from prevention to survivorship, to inform cancer control efforts. These surveys can illuminate cancer disparities among specific populations, including rural communities. However, due to small rural sample sizes, varying sampling methods, and/or other study design or analytical concerns, there are challenges in using population-based surveys for rural cancer control research and practice. Our objective is three-fold. First, we examined the characterization of "rural" in four, population-based surveys commonly referenced in the literature: 1) Health Information National Trends Survey (HINTS); 2) National Health Interview Survey (NHIS); 3) Behavioral Risk Factor Surveillance System (BRFSS); and 4) Medical Expenditures Panel Survey (MEPS). Second, we identified and described the challenges of using these surveys in rural cancer studies. Third, we proposed solutions to address these challenges. We found that these surveys varied in use of rural-urban classifications, sampling methodology, and available cancer-related variables. Further, we found that accessibility of these data to non-federal researchers has changed over time. Survey data have become restricted based on small numbers (i.e., BRFSS) and have made rural-urban measures only available for analysis at Research Data Centers (i.e., NHIS and MEPS). Additionally, studies that used these surveys reported varying proportions of rural participants with noted limitations in sufficient representation of rural minorities and/or cancer survivors. In order to mitigate these challenges, we propose two solutions: 1) make rural-urban measures more accessible to non-federal researchers and 2) implement sampling approaches to oversample rural populations.


Blockade of Central Angiotensin II AT1 Receptor Protects the Brain from Ischemia/Reperfusion Injury in Normotensive Rats.
Stroke is the third leading cause of invalidism and death in industrialized countries. There are conflicting reports about the effects of Angiotensin II on ischemia-reperfusion brain injuries and most data have come from chronic hypertensive rats. In this study, hypotensive and non-hypotensive doses of candesartan were used to investigate the effects of angiotensin II AT1 receptor blockade by transient focal cerebral ischemia in normotensive rats. In this experimental study, 48 male Sprague-Dawley rats were randomly divided into four groups (n=12). Sham group, the control ischemic group, and two ischemic groups received candesartan at doses of 0.1 or 0.5 mg/kg at one hour before ischemia. Transient focal cerebral ischemia was induced by 60 minutes occlusion of the middle cerebral artery, followed by 24 h reperfusion. The neurological deficit score was evaluated at the end of the reperfusion period. The total cortical and striatal infarct volumes were determined using triphenyltetrazolium chloride staining technique. Tissue swelling was calculated for the investigation of ischemic brain edema formation. In comparison with the control ischemic group, AT1 receptor blockade with both doses of candesartan (0.1 or 0.5 mg/kg) significantly improved neurological deficit and lowered cortical and striatal infarct sizes. In addition, pretreatment with candesartan significantly reduced ischemia induced tissue swelling. Angiotensin II by stimulating AT1 receptors, participates in ischemia-reperfusion injuries and edema formation. AT1 receptor blockade with candesartan decreased ischemic brain injury and edema and improved neurological outcome.


Activity-regulated cytoskeleton-associated protein/activity-regulated gene 3.1 (Arc/Arg3.1) enhances dendritic cell vaccination in experimental melanoma.
Dendritic cell (DC) vaccination has proven to be an effective and safe adjuvant for cancer immunotherapies. As the presence of DCs within the tumor microenvironment promotes adaptive antitumor immunity, enhancement of DC migration toward the tumor microenvironment following DC vaccination might represent one possible approach to increase its therapeutic efficacy. While recent findings suggest the activity-regulated cytoskeleton-associated protein/activity-regulated gene 3.1 (Arc/Arg3.1) as critical regulator of DC migration in the context of autoimmune diseases, we aimed to investigate the impact of Arc/Arg3.1 expression for DC-based cancer vaccines. To this end, DC migration capacity as well as the induction of T cell-mediated antitumor immunity was assessed in an experimental B16 melanoma model with Arc/Arg3.1-/- and Arc/Arg3.1-expressing BMDCs applied as a subcutaneous vaccine. While antigen presentation on DCs was critical for unleashing effective T cell mediated antitumor immune responses, Arc/Arg3.1 expression enhanced DC migration toward the tumor and secondary lymphoid organs. Moreover, Arc/Arg3.1-expressing BMDCs shape the tumor immune microenvironment by facilitating tumor recruitment of antigen-specific effector T cells. Thus, Arc/Arg3.1 may represent a novel therapeutic target in DCs in order to increase the therapeutic efficacy of DC vaccination.


Environmental Enrichment Upregulates Striatal Synaptic Vesicle-Associated Proteins and Improves Motor Function.
Environmental enrichment (EE) is a therapeutic paradigm that consists of complex combinations of physical, cognitive, and social stimuli. The mechanisms underlying EE-mediated synaptic plasticity have yet to be fully elucidated. In this study, we investigated the effects of EE on synaptic vesicle-associated proteins and whether the expression of these proteins is related to behavioral outcomes. A total of 44 CD-1® (ICR) mice aged 6 weeks were randomly assigned to either standard cages or EE (N = 22 each). Rotarod and ladder walking tests were then performed to evaluate motor function. To identify the molecular mechanisms underlying the effects of EE, we assessed differentially expressed proteins (DEPs) in the striatum by proteomic analysis. Quantitative real-time polymerase chain reaction (qRT-PCR), western blot, and immunohistochemistry were conducted to validate the expressions of these proteins. In the behavioral assessment, EE significantly enhanced performance on the rotarod and ladder walking tests. A total of 116 DEPs (54 upregulated and 62 downregulated proteins) were identified in mice exposed to EE. Gene ontology (GO) analysis demonstrated that the upregulated proteins in EE mice were primarily related to biological processes of synaptic vesicle transport and exocytosis. The GO terms for these biological processes commonly included Synaptic vesicle glycoprotein 2B (SV2B), Rabphilin-3A, and Piccolo. The qRT-PCR and western blot analyses revealed that EE increased the expression of SV2B, Rabphilin-3A and Piccolo in the striatum compared to the control group. Immunohistochemistry showed that the density of Piccolo in the vicinity of the subventricular zone was significantly increased in the EE mice compared with control mice. In conclusion, EE upregulates proteins associated with synaptic vesicle transport and exocytosis such as SV2B, Rabphilin-3A and Piccolo in the striatum. These upregulated proteins may be responsible for locomotor performance improvement, as shown in rotarod and ladder walking tests. Elucidation of these changes in synaptic protein expression provides new insights into the mechanism and potential role of EE.


Can nurses' shift work jeopardize the patient safety? A systematic review.
Medication administration accounts for 40% of the nursing clinical activity in hospitals and nurses play a central role in granting the patient safety, as they are directly responsible for the patient care. This review aims at analyzing the correlation between the clinical risk management and the occurrence of medication errors and the effects of the shift work (such as excessive fatigue and sleep deprivation after a shift in hospital) on inpatient nurses. This paper adheres to the relevant EQUATOR guidelines. A systematic review was conducted according to the PRISMA statement and pertinent articles were selected based on inclusion criteria and quality assessment factors. Two reviewers searched the bibliographic databases PubMed, Scopus, Cochrane, CINAHL to collect all the available articles in English and Italian issued between 1992 and August 2017. The reviewers analyzed 19 of the 723 initially extracted references, as they focused on the impact of workload, shifts and sleep deprivation on the probability of making medication errors. The main reasons behind medication errors are stress, fatigue, increased workload, night shifts, nurse staffing ratio and workflow interruptions. These factors can have a significant negative impact on the health and the performance of the employees. It is desirable to extend and deepen the research to identify appropriate measures to minimize medication errors.


Cognitive ability at kindergarten entry and socioeconomic status.
To examine how gradients in socioeconomic status (SES) impact US children's reading and math ability at kindergarten entry and determine the contributions of family background, health, home learning, parenting, and early education factors to those gradients. Analysis of 6600 children with cognitive assessments at kindergarten entry from the US Early Childhood Longitudinal Birth Cohort Study. A composite SES measure based on parent's occupation, education, and income was divided into quintiles. Wald F tests assessed bivariate associations between SES and child's cognitive ability and candidate explanatory variables. A decomposition methodology examined mediators of early cognitive gradients. Average reading percentile rankings increased from 34 to 67 across SES quintiles and math from 33 to 70. Children in lower SES quintiles had younger mothers, less frequent parent reading, less home computer use (27%-84%), and fewer books at home (26-114). Parent's supportive interactions, expectations for their child to earn a college degree (57%-96%), and child's preschool attendance (64%-89%) increased across quintiles. Candidate explanatory factors explained just over half the gradients, with family background factors explaining 8% to 13%, health factors 4% to 6%, home learning environment 18%, parenting style/beliefs 14% to 15%, and early education 6% to 7% of the gaps between the lowest versus highest quintiles in reading and math. Steep social gradients in cognitive outcomes at kindergarten are due to many factors. Findings suggest policies targeting levels of socioeconomic inequality and a range of early childhood interventions are needed to address these disparities.


Rectal analgesia for pain from perineal trauma following childbirth.
Perineal pain from a tear and/or surgical cut (episiotomy) is a common problem following vaginal birth. Strategies to reduce perineal trauma and the appropriate repair of any perineal damage sustained are important for avoiding and alleviating pain. Where pain is present, numerous treatments are used in clinical practice, such as local anaesthetics, oral analgesics, therapeutic ultrasound, antiseptics and non-pharmacological applications such as ice packs and baths. This review assesses the evidence for using rectal analgesia for pain relief following perineal trauma. To assess the effectiveness of analgesic rectal suppositories for pain from perineal trauma following childbirth. We searched the Cochrane Pregnancy and Childbirth Group trials register (July 2002), CENTRAL (The Cochrane Library, Issue 2, 2002), CINAHL (May 2002) and MIDIRS (May 2002). Randomised controlled trials comparing analgesic rectal suppositories with placebo or alternative treatment for the relief of perineal pain. Two reviewers assessed trial quality and extracted data independently. Three trials involving 249 women met the inclusion criteria. Only two of the trials identified for inclusion in this review had data that could be entered in a meta-analysis, with the third not providing data in a useable format. Women were less likely to experience pain at or close to 24 hours after birth if they received non-steroidal anti-inflammatory drugs (NSAID) suppositories compared with placebo (relative risk (RR) 0.37, 95% confidence interval (CI) 0.10 to 1.38, 2 trials, 150 women). Women in the NSAID suppositories group compared with women in the placebo group required less additional analgesia in the first 24 hours after birth (RR 0.31, 95% CI 0.17 to 0.54, 1 trial, 89 women) and this effect was still evident at 48 hours postpartum (RR 0.63, 95% CI 0.45 to 0.89, 1 trial, 89 women). No information was available on pain experienced more than 72 hours after birth or other outcomes of importance to women such as the impact on daily activities, resumption of sexual intercourse and the impact on the mother-baby relationship. NSAID rectal suppositories are associated with less pain up to 24 hours after birth, and less additional analgesia is required. More research is required regarding long-term effects and maternal satisfaction with the treatment.


Mechanism of glutathione transferase P1-1-catalyzed activation of the prodrug canfosfamide (TLK286, TELCYTA).
Canfosfamide (TLK286, TELCYTA) is a prodrug that upon activation by glutathione transferase P1-1 (GST P1-1) yields an anticancer alkylating agent and a glutathione derivative. The rationale underlying the use of TLK286 in chemotherapy is that tumor cells overexpressing GST P1-1 will be locally exposed to the released alkylating agent with limited collateral toxicity to the surrounding normal tissues. TLK286 has demonstrated clinical effects in phase II and III clinical trials for the treatment of malignancies, such as ovarian cancer, nonsmall cell lung cancer, and breast cancer, as a single agent and in combination with other chemotherapeutic agents. In spite of these promising results, the detailed mechanism of GST P1-1 activation of the prodrug has not been elucidated. Here, we propose a mechanism for the TLK286 activation by GST P1-1 on the basis of density functional theory (DFT) and on potential of mean force (PMF) calculations. A catalytic water molecule is instrumental to the activation by forming a network of intermolecular interactions between the active-site Tyr7 hydroxyl and the sulfone and COO(-) groups of TLK286. The results obtained are consistent with the available experimental kinetic data and provide an atomistic understanding of the TLK286 activation mechanism.


Edoxaban for treatment of venous thromboembolism in patients with cancer. Rationale and design of the Hokusai VTE-cancer study.
Direct oral anticoagulants may be effective and safe for treatment of venous thromboembolism (VTE) in cancer patients, but they have not been compared with low-molecular-weight heparin (LMWH), the current recommended treatment for these patients. The Hokusai VTE-cancer study is a randomised, open-label, clinical trial to evaluate whether edoxaban, an oral factor Xa inhibitor, is non-inferior to LMWH for treatment of VTE in patients with cancer. We present the rationale and some design features of the study. One such feature is the composite primary outcome of recurrent VTE and major bleeding during a 12-month study period. These two complications occur frequently in cancer patients receiving anticoagulant treatment and have a significant impact. The evaluation beyond six months will fill the current gap in the evidence base for the long-term treatment of these patients. Based on the observation that the risk of recurrent VTE in patients with active cancer is similar to that in those with a history of cancer, the Hokusai VTE-cancer study will enrol patients if whose cancer was diagnosed within the past two years. In addition, patients with incidental VTE are eligible because their risk of recurrent VTE is similar to that in patients with symptomatic disease. The unique design features of the Hokusai VTE-cancer study should lead to enrolment of a broad spectrum of cancer patients with VTE who could benefit from oral anticoagulant treatment.


Capping a pulpotomy with calcium aluminosilicate cement: comparison to mineral trioxide aggregates.
Calcium aluminate cements have shown little affinity for bacterial growth, low toxicity, and immunogenicity when used as a restoration material, but calcium aluminate cements have not been tested in vivo in pulpotomy procedures. To address this question, a calcium aluminosilicate cement (Quick-Set) was tested along with 2 mineral trioxide aggregates, ProRoot MTA and MTA Plus. These cements were used as a capping agent after pulpotomy. Control rats had no pulpotomy, or the pulpotomy was not capped. Proinflammatory cytokines interleukin (IL)-1β and IL-1α were measured, and histology was performed at 30 and 60 days after capping. The nociceptive response was determined by measuring the lengthening of the rat's meal duration. and IL-1β and IL-1α concentrations were reduced in the capped teeth, but no differences were observed among the 3 cements. Dentinal bridging could be detected at both 30 and 60 days with each of the 3 cements, and the pulps were still vital 60 days after capping. Meal duration significantly shortened after placement of the 3 different cements, indicating a nociceptive response, but there were no differences among the materials. Calcium aluminosilicate cement had similar properties to mineral trioxide aggregates and is a viable option for pulpotomy procedures.


Purification, partial characterization and biological effects of the XTC mesoderm-inducing factor.
The mesoderm of Xenopus laevis is formed through an inductive interaction in which a signal from the vegetal hemisphere of the blastula acts on overlying animal pole cells. We have recently reported that the Xenopus XTC cell line secretes a mesoderm-inducing factor (MIF) which may resemble the natural signal. In this paper, we describe the purification and biological effects of XTC-MIF. XTC-MIF is a hydrophobic protein with an isoelectric point of 7.8 and an apparent relative molecular mass (Mr) of 23,500. On reduction, XTC-MIF loses its biological activity and the protein dissociates into two inactive subunits with apparent Mr of about 15,000. These properties closely resemble those of transforming growth factor type beta (TGF-beta), and it is interesting that TGF-beta 2 has recently been shown to have mesoderm-inducing activity. The biological response to XTC-MIF is graded. After exposure to 0.2-1.0 ng ml-1 XTC-MIF, stage-8 animal pole explants form mesenchyme and mesothelium. At higher concentrations, up to about 5 ng ml-1, muscle is formed, occasionally with neural tissue. In response to concentrations of XTC-MIF greater than 5-10 ng ml-1, notochord and neural tissue are usually formed. The formation of notochord and neural tissue in response to XTC-MIF represents a qualitative difference between this inducing factor and the other known group of MIFs, the heparin-binding growth factors.


General tests of the Markov property in multi-state models.
Multi-state models for event history analysis most commonly assume the process is Markov. This article considers tests of the Markov assumption that are applicable to general multi-state models. Two approaches using existing methodology are considered; a simple method based on including time of entry into each state as a covariate in Cox models for the transition intensities and a method involving detecting a shared frailty through a stratified Commenges-Andersen test. In addition, using the principle that under a Markov process the future rate of transitions of the process at times $t > s$ should not be influenced by the state occupied at time $s$, a new class of general tests is developed by considering summaries from families of log-rank statistics where patients are grouped by the state occupied at varying initial time $s$. An extended form of the test applicable to models that are Markov conditional on observed covariates is also derived. The null distribution of the proposed test statistics are approximated by using wild bootstrap sampling. The approaches are compared in simulation and applied to a dataset on sleeping behavior. The most powerful test depends on the particular departure from a Markov process, although the Cox-based method maintained good power in a wide range of scenarios. The proposed class of log-rank statistic based tests are most useful in situations where the non-Markov behavior does not persist, or is not uniform in nature across patient time.


DNA replication is the target for the antibacterial effects of nonsteroidal anti-inflammatory drugs.
Evidence suggests that some nonsteroidal anti-inflammatory drugs (NSAIDs) possess antibacterial properties with an unknown mechanism. We describe the in vitro antibacterial properties of the NSAIDs carprofen, bromfenac, and vedaprofen, and show that these NSAIDs inhibit the Escherichia coli DNA polymerase III β subunit, an essential interaction hub that acts as a mobile tether on DNA for many essential partner proteins in DNA replication and repair. Crystal structures show that the three NSAIDs bind to the sliding clamp at a common binding site required for partner binding. Inhibition of interaction of the clamp loader and/or the replicative polymerase α subunit with the sliding clamp is demonstrated using an in vitro DNA replication assay. NSAIDs thus present promising lead scaffolds for novel antibacterial agents targeting the sliding clamp.


Endocrine disrupting chemicals: effects on human male reproductive health.
There is now considerable evidence that male reproductive function is declining in human and wildlife populations. This is coincident with the increasing use and prevalence of man-made chemicals in the environment over the last fifty years. Certain chemicals have subsequently been shown to disturb the developing fetal endocrine system of laboratory animals in utero. In these experiments, treatment caused similar male reproductive problems in offspring as those already observed in wildlife and human populations. In addition, both the human DES data and rodent studies have shown that there are specific windows of gestation when the developing fetal gonad is highly sensitive to small endocrine changes. Animal in vivo and human in vitro studies have identified EDC sensitive genes. Consequently, hypotheses are being generated concerning mechanism of action e.g. disturbed testicular apoptosis and altered hepatic biotransformation of steroids. While animal studies provide us with valuable insights into the range of effects that can be attributed to in utero EDC exposure, varying maternal doses employed by different research groups make relation of the results to human observations difficult. The EDC concentration representative of fetal exposure levels is uncertain. Confounding factors include: (a) the vast number of chemicals termed EDCs, (b) the ability of chemicals to bioaccumulate in body lipid, (c) the metabolism of body lipid during pregnancy releasing the mothers lifetime EDC legacy into circulation and (d) the poorly understood kinetics of EDC transfer across the placenta. Thus, the level of fetal exposure can only be crudely estimated at present. This highlights the need for large animal models of EDC in utero exposure where the partitioning of EDCs between the mother and fetus and transfer across the placenta can be studied in detail. Despite considerable effort the mechanisms by which these endocrine disrupting chemicals exert their effects are still largely unknown. Further studies of the mechanism of action, and consequences, of EDCs in fetal development must be done in order to elucidate how EDCs exert their effects. This can only be achieved using a combined approach whereby animal models are used in combination with in vitro human studies. In conclusion however, there are now sufficient animal model data to prove that EDCs can adversely affect reproductive development and function in the male. Our further understanding of the mechanisms involved may allow intervention strategies whereby we can at least prevent a further decline in male as well as female reproductive health.