Restrictions of anthelmintic usage: perspectives and potential consequences.
Given the increasing levels of anthelmintic resistance in equine parasites, parasitologists now recommend traditional treatment approaches to be abandoned and replaced by more sustainable strategies. It is of crucial importance to facilitate veterinary involvement to ensure that treatment decisions are based on parasitic knowledge. Despite recommendations given for the past two decades, strategies based on the selective therapy principle have not yet been implemented on a larger scale in equine establishments. In contrast, treatment regimens appear to be derived from recommendations originally given in 1966. The province of Quebec in Canada, and an increasing number of European countries, have implemented prescription-only restrictions on anthelmintic drugs. Denmark introduced this legislation ten years ago, and some evidence has been generated describing potential consequences. It is without dispute that Danish veterinarians are now deeply involved with parasite management in equine establishments. However, little is known about the impact on levels of anthelmintic resistance and the risk of parasitic disease under these circumstances. In addition, the legislation makes huge demands on diagnosis and parasite surveillance. No data have been published evaluating fecal egg count techniques and larval culture methods as clinical diagnostic tools, and very little is known about potential correlations with actual worm burdens. This article provides a general review of anthelmintic strategies currently used in equine establishments and outlines the recommendations now given for parasite control. Preliminary experience with prescription-only restrictions in Denmark is presented and current research needs to further evaluate this approach are discussed.


HIV-1 envelope protein gp140 binding studies to human brain microvascular endothelial cells.
Human immunodeficiency virus type-1 (HIV-1) envelope glycoprotein gp140 interacts with its specific receptors on the surface of the target cells leading to cellular activation through various signaling pathways. The effect of blocking the chemokine repertoire in human brain microvascular endothelial cells in HIV dementia (HAD) disease has not been reported. Characterizing the nature of HIV-1 envelope protein gp140 (T-tropic, HXBc2) receptor binding conditions to HBMEC is critical to gain insight into the HIV dementia, and eventually to rationally design the agents to block envelope protein receptor interactions. HIV-1 gp140 oligomers were purified and separated to monomers, dimers, and trimers. The binding conditions of gp140 to HBMEC chemokine receptor, CXCR4, were optimized with an aim of understanding the structural interactions in HAD. Analysis of the interaction between HIV-1 gp140 and CXCR4 of HBMEC by saturation binding, cross-competition analysis with radiolabeled SDF and gp140, revealed a strong interaction, specificity between HIV-1 gp140 and CXCR4. Our binding data demonstrate that HIV-1 envelope protein gp140 enters cells by protein receptor mediated interactions that are regulated by the conformational state of the gp140 at physiological environment (pH and temperature). The CXCR4 antibody 12G5 inhibited SDF-1 binding to HBMEC indicating the specificity of gp140 binding to HBMEC. Scatchard analysis revealed the presence of approximately 70250 gp140 binding sites per cell with a K(d) of 4.5 nM. Cross-competition experiments using labeled SDF-1 and gp140 revealed that both unlabeled SDF-1 and gp140 are capable of displacing their radiolabeled counterparts. The binding assay conditions and radioligand binding assay are highly valuable to identify and design better HIV inhibitors for HAD.


Fgf10 is essential for limb and lung formation.
The interactions between fibroblast growth factors (FGF) and their receptors have important roles in mediating mesenchymal-epithelial cell interactions during embryogenesis. In particular, Fgf10 is predicted to function as a regulator of brain, lung and limb development on the basis of its spatiotemporal expression pattern in the developing embryo. To define the role of Fgf10, we generated Fgf10-deficient mice. Fgf10-/- mice died at birth due to the lack of lung development. Trachea was formed, but subsequent pulmonary branching morphogenesis was disrupted. In addition, mutant mice had complete truncation of the fore- and hindlimbs. In Fgf10-/- embryos, limb bud formation was initiated but outgrowth of the limb buds did not occur; however, formation of the clavicles was not affected. Analysis of the expression of marker genes in the mutant limb buds indicated that the apical ectodermal ridge (AER) and the zone of polarizing activity (ZPA) did not form. Thus, we show here that Fgf10 serves as an essential regulator of lung and limb formation.


Clinical evaluation of brief cognitive assessment measures for patients with severe dementia.
Alzheimer's disease has become an important public health burden for older adults. Clinicians face a challenging task to efficiently evaluate cognition in dementia in clinical settings. We sought to assess the validity and inter-correlations of brief cognitive assessments in a cohort of severely demented patients. In total, 49 individual patients (N = 49) ranging in age from 62 to 97 years old were included in this performance improvement project. Over the course of two-three sessions, five cognitive instruments were administered to each patient: Severe Impairment Battery (SIB), Severe Impairment Battery-8 (SIB-8), Mini Mental State Examination (MMSE), Severe Mini Mental State Examination (sMMSE) and Brief Interview of Mental Status (BIMS). We sought to assess patient factors that might have been barriers to optimal performance on cognitive/functional tests. Researchers assessed her impression of the participants' difficulty comprehending instructions, distractibility, apparent fatigue, and frustration, which were the four barriers rated. Data were analyzed for 49 patients from the inpatient dementia unit with a total of 51 samples. All of the inter-correlations between the five cognitive instruments had Spearman coefficients of (rs) > 0.7 and were statistically significant with p < 0.001. The SIB-8 and sMMSE were positively correlated with the SIB. The perceived barrier scores ranged from 0- no issue to 1-mild issue on all five cognitive instruments. Brief cognitive tests designed for severe dementia such as the SIB-8 and sMMSE have been evaluated in this project to be shorter in administration duration and highly correlated with gold standard instruments: the SIB and MMSE.


Custom designing therapeutic cancer vaccines: delivery of immunostimulatory molecule adjuvants by protein transfer.
Attempts to create vaccines for humans against invading pathogens such as viruses and bacteria have met with tremendous success. The process of developing vaccines against these pathogens is greatly aided by the fact that they contain antigens that are entirely foreign to humans. Although the knowledge and strategies developed for designing vaccines against these microbes may be of use in developing cancer vaccines, the poor antigenicity and immunosuppressive ability of cancers pose major hurdles to vaccine development. Established tumors have not only withstood immune screening and selection pressure, making them poor stimulators of an immune response, but have also adapted mechanisms to continue evading immune surveillance by creating an immunosuppressive environment. Also, genetic differences in immune responses to an antigen among individuals result in an antigenic profile that varies from patient to patient. Cancers bear such great similarities to normal cells in the body that, on a molecular level, the differences between cancerous and non-cancerous cells are minor. Therefore, developing vaccines which use the host's own tumor tissues carries the risk of breaking tolerance to self-antigens that are present in the tumor tissue. Vaccination strategies that will optimally stimulate the immune system against tumor specific antigens under immunosuppressive conditions need to be developed. In practical terms, this calls for a method by which therapeutic vaccines may be custom-designed to treat cancers case by case. Ex vivo manipulation of dendritic cells and gene transfer of immunostimulatory molecules in ex vivo expanded tumors are being tested in both experimental models and also in human clinical trials. Some of them have met with limited success. Emerging technologies such as protein transfer, which make it possible to express immunostimulatory molecules on tumor cell membranes, offer the means to develop efficient tumor vaccines that are simple and fast, while being easy to store and administer in human patients. Progress in these techniques will move the cancer vaccine field a step closer towards realizing custom designed cancer vaccines in human clinical settings.


Statistical analysis of molecule colocalization in bioimaging.
The quantitative analysis of molecule interactions in bioimaging is key for understanding the molecular orchestration of cellular processes and is generally achieved through the study of the spatial colocalization between the different populations of molecules. Colocalization methods are traditionally divided into pixel-based methods that measure global correlation coefficients from the overlap between pixel intensities in different color channels, and object-based methods that first segment molecule spots and then analyze their spatial distributions with second-order statistics. Here, we present a review of such colocalization methods and give a quantitative comparison of their relative merits in different types of biological applications and contexts. We show on synthetic and biological images that object-based methods are more robust statistically than pixel-based methods, and allow moreover to quantify accurately the number of colocalized molecules.


Time course of adaptive regulatory peptide changes following massive small bowel resection in the dog.
Basal and postprandial concentrations of gastrointestinal hormones were measured in 12 dogs before and at one and three months after a 75% small bowel resection. Five animals were studied again at six months. Concentrations of enteric hormones and neuropeptides, measured in the proximal jejunum and distal ileum adjacent to the anastomotic site at the time of euthanasia, were compared with concentrations in control tissues taken from each animal at the time of resection. Increased basal and postprandial levels of gastrin (P < 0.05), cholecystokinin (CCK, P < 0.05), glucose-dependent insulinotropic peptide (GIP, P < 0.01), peptide YY (PYY, P < 0.001), and enteroglucagon (P < 0.001), were seen at one month after small bowel resection. In contrast, no significant changes were seen in concentrations of secretin, motilin, neurotensin, somatostatin, PP, or glucagon. Concentrations of enteroglucagon, GIP, and PYY remained high throughout the six-month study period. In contrast, gastrin and CCK had normalized by three months. Thus, only enteroglucagon, PYY, and GIP showed sustained elevations following enterectomy; the gastrin and CCK changes were transient. Following enterectomy, concentrations of vasoactive intestinal polypeptide (VIP) were reduced by about 50% in mucosal (P < 0.001) and muscle (P < 0.05) layers of proximal and distal gut. In contrast, calcitonin gene-related peptide (CGRP) was increased by about twofold in jejunal and ileal mucosa (P < 0.05), and CGRP elevations were even more marked in the muscle layers (P < 0.001). Somatostatin and neuropeptide Y (NPY) concentrations were similar to controls in all areas except for a small decrease in NPY in ileal mucosa (P < 0.05). These findings suggest that the increased motilin and PP concentrations previously reported after bowel resection in man are more likely to reflect underlying inflammatory bowel disease rather than enterectomy. The normalization of hypergastrinemia explains why the increased acid secretion after small bowel resection is transient. These results provide evidence for independent secretory control of enteroglucagon and PYY, which are both products of intestinal L cells. In addition, these studies reveal marked changes in enteric neuropeptide concentrations following bowel resection. VIP, which is thought to be a major inhibitory transmitter in the gut, is markedly reduced, while CGRP, which is mainly localized in sensory afferent fibers, is increased. These major neuropeptide changes are likely to be of importance in the adaptive responses to massive small bowel resection.


Comparison of spirometric reference values.
Lung-function reference values play a vital role in the management of respiratory disorders. There are many proposed reference equations for pediatric spirometry. Recently, spirometric reference equations were proposed, using data from people aged 8-80 years living in the US compiled by the third National Health and Nutrition Examination Survey. Our objective was to compare the predictive value of wider age-range reference equations to established pediatric reference equations for the pediatric population. Spirometry, height, and weight were obtained from 70 normal children aged 6-18 years. The difference between measured and predicted values as suggested by different reference equations was compared. Predicted values from general equations significantly differed from those generated from pediatric equations and from measured values in this population. The difference between measured and predicted values from the wider age-range equations varied between 7-16% for forced expired volume in 1 sec (FEV1) and forced vital capacity (FVC). The difference between measured and predicted values for the pediatric equations varied between 1-4%. Although wider age-range equations provide continuity through age ranges, their predictive accuracy may be low in the pediatric age group, especially for the youngest, smallest children. Extrapolating reference equations beyond the age range of subjects used to generate then is not recommended.


Media coverage of calls to rename low-risk cancers: a content analysis.
The use of more medicalised labels can increase both concern about illness and the desire for more invasive treatment. This study analyses the media's coverage of an Analysis article in The BMJ which generated a large amount of high-profile international media coverage. It aims to understand how to better communicate messages about low-risk cancers and overdiagnosis to the public. Content analysis of media coverage. Media was identified by Isentia Media Portal, searched in Google News and cross-checked in Factiva and Proquest databases from August 2018. Media headlines, full text and open access public comments responding to the coverage on the article proposing to 'rename low-risk conditions currently labelled as cancer' were analysed to determine the main themes. 45 original media articles and their associated public comments (n=167) were identified and included in the analysis. Overall, headlines focused on cancer generally and there was little mention of 'low-risk', 'overdiagnosis' or 'overtreatment'. The full text generally presented a more balanced view of the evidence and were supportive of the proposal, however, public responses tended to be more negative towards the idea of renaming low-risk cancers and indicated confusion. Comments seemed to focus on the headlines rather than the full article. This study offers a novel insight into media coverage of the complex and counterintuitive problem of overdiagnosis. Continued deliberation on how to communicate similar topics to the public through the mainstream media is needed. Future work in the area of low-risk cancer communication should consider the powerful impact of people's previous experience with a cancer diagnosis and the criticism about being paternalistic and concealing the truth from patients.


Raf-induced transformation requires an interleukin 1 autocrine loop.
The c-Raf-1 serine/threonine protein kinase plays a critical role in the proliferation of most cell types that have been examined. As such, the Raf proto-oncogene is thought to play a central role in the development of human tumors. Although the c-raf-1 gene itself rarely appears to be mutated in human tumors, the kinase activity of Raf is frequently found to be more active in tumor cells, likely through constitutive activation of upstream activators of Raf. The downstream events triggered by Raf that are involved in transformation have been studied less extensively. We show in this study that Raf-induced transformation of NIH 3T3 cells requires the activation of the ubiquitously expressed transcription factor, nuclear factor-kappaB, by Raf. Furthermore, through the use of CrmA, interleukin 1 (IL-1) receptor antagonist, and a dominant-negative form of TRAF6, we demonstrate a requirement for IL-1 production and signaling from the IL-1 receptor as necessary components of Raf-induced transformation. These results indicate that IL-1 may be used as an autocrine growth factor by a number of tumors in which activation of Raf plays an important role in transformation and suggest that blockade of IL-1 signaling may be an approach to limiting the growth of certain tumors.


Colitis-induced colorectal cancer and intestinal epithelial estrogen receptor beta impact gut microbiota diversity.
Chronic inflammation of the colon (colitis) is a risk factor for colorectal cancer (CRC). Hormone-replacement therapy reduces CRC incidences, and the estrogen receptor beta (ERβ/ESR2) has been implicated in this protection. Gut microbiota is altered in both colitis and CRC and may influence the severity of both. Here we test the hypothesis that intestinal ERβ impacts the gut microbiota. Mice with and without intestine-specific deletion of ERβ (ERβKOVil ) were generated using the Cre-LoxP system. Colitis and CRC were induced with a single intraperitoneal injection of azoxymethane (AOM) followed by administration of three cycles of dextran sulfate sodium (DSS) in drinking water. The microbiota population were characterized by high-throughput 16S rRNA gene sequencing of DNA extracted from fecal samples (N = 39). Differences in the microbiota due to AOM/DSS and absence of ERβ were identified through bioinformatic analyses of the 16S-Seq data, and the distribution of bacterial species was corroborated using qPCR. We demonstrate that colitis-induced CRC reduced the gut microbiota diversity and that loss of ERβ enhanced this process. Further, the Bacteroidetes genus Prevotellaceae_UCG_001 was overrepresented in AOM/DSS mice compared to untreated controls (3.5-fold, p = 0.004), and this was enhanced in females and in ERβKOVil mice. Overall, AOM/DSS enriched for microbiota impacting immune system diseases and metabolic functions, and lack of ERβ in combination with AOM/DSS enriched for microbiota impacting carbohydrate metabolism and cell motility, while reducing those impacting the endocrine system. Our data support that intestinal ERβ contributes to a more favorable microbiome that could attenuate CRC development.


A Mechanistic Physiologically-Based Biopharmaceutics Modeling (PBBM) Approach to Assess the In Vivo Performance of an Orally Administered Drug Product: From IVIVC to IVIVP.
The application of in silico modeling to predict the in vivo outcome of an oral drug product is gaining a lot of interest. Fully relying on these models as a surrogate tool requires continuous optimization and validation. To do so, intraluminal and systemic data are desirable to judge the predicted outcomes. The aim of this study was to predict the systemic concentrations of ibuprofen after oral administration of an 800 mg immediate-release (IR) tablet to healthy subjects in fasted-state conditions. A mechanistic oral absorption model coupled with a two-compartmental pharmacokinetic (PK) model was built in Phoenix WinNonlinWinNonlin® software and in the GastroPlus™ simulator. It should be noted that all simulations were performed in an ideal framework as we were in possession of a plethora of in vivo data (e.g., motility, pH, luminal and systemic concentrations) in order to evaluate and optimize these models. All this work refers to the fact that important, yet crucial, gastrointestinal (GI) variables should be integrated into biopredictive dissolution testing (low buffer capacity media, considering phosphate versus bicarbonate buffer, hydrodynamics) to account for a valuable input for physiologically-based pharmacokinetic (PBPK) platform programs. While simulations can be performed and mechanistic insights can be gained from such simulations from current software, we need to move from correlations to predictions (IVIVC → IVIVP) and, moreover, we need to further determine the dynamics of the GI variables controlling the dosage form transit, disintegration, dissolution, absorption and metabolism along the human GI tract. Establishing the link between biopredictive in vitro dissolution testing and mechanistic oral absorption modeling (i.e., physiologically-based biopharmaceutics modeling (PBBM)) creates an opportunity to potentially request biowaivers in the near future for orally administered drug products, regardless of its classification according to the Biopharmaceutics Classification System (BCS).


Dolichoectatic aneurysms of the vertebrobasilar system: clinical and radiographic factors that predict poor outcomes.
OBJECTIVE Fusiform dolichoectatic vertebrobasilar aneurysms are rare, challenging lesions. The natural history of these lesions and medium- and long-term patient outcomes are poorly understood. The authors sought to evaluate patient prognosis after diagnosis of fusiform dolichoectatic vertebrobasilar aneurysms and to identify clinical and radiographic predictors of neurological deterioration. METHODS The authors reviewed multiple, prospectively maintained, single-provider databases at 3 large-volume cerebrovascular centers to obtain data on patients with unruptured, fusiform, basilar artery dolichoectatic aneurysms diagnosed between January 1, 2000, and January 1, 2015. RESULTS A total of 50 patients (33 men, 17 women) were identified; mean clinical follow-up was 50.1 months and mean radiographic follow-up was 32.4 months. At last follow-up, 42% (n = 21) of aneurysms had progressed and 44% (n = 22) of patients had deterioration of their modified Rankin Scale scores. When patients were dichotomized into 2 groups- those who worsened and those who did not-univariate analysis showed 5 variables to be statistically significantly different: sex (p = 0.007), radiographic brainstem compression (p = 0.03), clinical posterior fossa compression (p < 0.001), aneurysmal growth on subsequent imaging (p = 0.001), and surgical therapy (p = 0.006). A binary logistic regression was then created to evaluate these variables. The only variable found to be a statistically significant predictor of clinical worsening was clinical symptoms of posterior fossa compression at presentation (p = 0.01). CONCLUSIONS Fusiform dolichoectatic vertebrobasilar aneurysms carry a poor prognosis, with approximately one-half of the patients deteriorating or experiencing progression of their aneurysm within 5 years. Despite being high risk, intervention-when carefully timed (before neurological decline)-may be beneficial in select patients.


Oncostatin M enhances the antiviral effects of type I interferon and activates immunostimulatory functions in liver epithelial cells.
Oncostatin M (OSM) is released together with type I interferon (IFN) by activated dendritic cells, suggesting a concerted action of these cytokines in the biological response against infection. We found that OSM increases the antiviral effect of IFN-alpha in Huh7 hepatoma cells infected with hepatitis A or hepatitis C virus and synergizes with IFN-alpha in the induction of antiviral genes. The combination of OSM and IFN-alpha led to upregulation of both STAT1 and STAT3 together with intense and prolonged activation of STAT1, STAT3, and Jak1. OSM with or without IFN-alpha also activated p38 mitogen-activated protein kinase, which is known to enhance transcription of IFN-alpha-inducible genes. Interestingly, OSM combined with IFN-alpha strongly induced immunoproteasome genes and other genes involved in antigen processing and presentation. Moreover, OSM, alone or in combination with IFN-alpha, upregulated relevant innate immunity molecules and increased the expression of intracellular adhesion molecule 1 and interleukin-15 receptor alpha (IL-15Ralpha) in liver cells. Hepatoma cells transfected with a plasmid encoding a viral antigen were able to activate effector T cells when pretreated with IFN-alpha plus OSM but not with each cytokine separately. Also, OSM, more than IFN-alpha, augmented the ability of Huh7 cells to transpresent IL-15 to responding lymphocytes and increased the immunostimulatory activity of liver epithelial cells by presenting a short viral peptide to sensitized cytotoxic T cells. In conclusion, OSM enhances the antiviral effects of type I interferon and cooperates with it in the induction of adaptive immune responses to pathogens. These findings may have therapeutic implications.


Varicella zoster virus vaccines: potential complications and possible improvements.
Varicella zoster virus (VZV) is the causative agent of varicella (chicken pox) and herpes zoster (shingles). After primary infection, the virus remains latent in sensory ganglia, and reactivates upon weakening of the cellular immune system due to various conditions, erupting from sensory neurons and infecting the corresponding skin tissue. The current varicella vaccine (v-Oka) is highly attenuated in the skin, yet retains its neurovirulence and may reactivate and damage sensory neurons. The reactivation is sometimes associated with postherpetic neuralgia (PHN), a severe pain along the affected sensory nerves that can linger for years, even after the herpetic rash resolves. In addition to the older population that develops a secondary infection resulting in herpes zoster, childhood breakthrough herpes zoster affects a small population of vaccinated children. There is a great need for a neuro-attenuated vaccine that would prevent not only the varicella manifestation, but, more importantly, any establishment of latency, and therefore herpes zoster. The development of a genetically-defined live-attenuated VZV vaccine that prevents neuronal and latent infection, in addition to primary varicella, is imperative for eventual eradication of VZV, and, if fully understood, has vast implications for many related herpesviruses and other viruses with similar pathogenic mechanisms.


Anterior cruciate ligament reconstruction: a multicenter prospective cohort study evaluating 3 different grafts using same bone drilling method.
To compare the clinical outcomes after anterior cruciate ligament (ACL) reconstruction with bone-patellar tendon-bone autograft (BPTBAu), BPTB allograft (BPTBAll), or hamstring (semitendinosus-gracilis) tendon autograft (HTAu), performing bone drilling with same methods in terms of transtibial drilling, orientation, positioning, and width of femoral and tibial tunnels. Multicenter prospective cohort study (level of evidence II). Departments of Orthopedic Surgery of Centro Médico Teknon (Barcelona, Spain) Clínica Universitaria de Navarra (Navarra, Spain), and Clínica FREMAP (Gijón, Spain). All patients with ACL tears attending 3 different institutions between January 2004 and June 2006 were approached for eligibility and those meeting inclusion criteria finally participated in this study. Each institution was assigned to perform a specific surgical technique. Patients were prospectively followed after undergoing ACL reconstruction with BPTBAu, BPTBAll, or HTAu, with a minimum follow-up of 24 months. Included knee laxity and International Knee Documentation Committee (IKDC) score. Knee laxity was assessed with the KT-1000 arthrometer (evaluated with neutral and external rotation positions) and both Lachman and pivot shift tests. Additional outcomes included main symptoms (anterior knee pain, swelling, crepitation, and instability), disturbance in knee sensation, visual analogue scale (VAS) for satisfaction with surgery, range of motion (ROM), and isokinetic knee strength. There were no significant differences among the 3 groups for any of the clinical outcomes, except for a slightly greater KT-1000-measured knee laxity in external rotation in the BPTBAu compared with the other groups. All patients demonstrated grade A or B of the IKDC. The mean VAS for satisfaction with surgery in all patients was 8.5. The selection of the surgical technique for ACL reconstruction may be based on the surgeon's preferences.


Identification of genetic determinants of breast cancer immune phenotypes by integrative genome-scale analysis.
Cancer immunotherapy is revolutionizing the clinical management of several tumors, but has demonstrated limited activity in breast cancer. The development of more effective treatments is hindered by incomplete knowledge of the genetic determinant of immune responsiveness. To fill this gap, we mined copy number alteration, somatic mutation, and expression data from The Cancer Genome Atlas (TCGA). By using RNA-sequencing data from 1,004 breast cancers, we defined distinct immune phenotypes characterized by progressive expression of transcripts previously associated with immune-mediated rejection. The T helper 1 (Th-1) phenotype (ICR4), which also displays upregulation of immune-regulatory transcripts such as PDL1, PD1, FOXP3, IDO1, and CTLA4, was associated with prolonged patients' survival. We validated these findings in an independent meta-cohort of 1,954 breast cancer gene expression data. Chromosome segment 4q21, which includes genes encoding for the Th-1 chemokines CXCL9-11, was significantly amplified only in the immune favorable phenotype (ICR4). The mutation and neoantigen load progressively decreased from ICR4 to ICR1 but could not fully explain immune phenotypic differences. Mutations of TP53 were enriched in the immune favorable phenotype (ICR4). Conversely, the presence of MAP3K1 and MAP2K4 mutations were tightly associated with an immune-unfavorable phenotype (ICR1). Using both the TCGA and the validation dataset, the degree of MAPK deregulation segregates breast tumors according to their immune disposition. These findings suggest that mutation-driven perturbations of MAPK pathways are linked to the negative regulation of intratumoral immune response in breast cancer. Modulations of MAPK pathways could be experimentally tested to enhance breast cancer immune sensitivity.


Assessing impact of physical activity-based youth development programs: validation of the Life Skills Transfer Survey (LSTS).
A signature characteristic of positive youth development (PYD) programs is the opportunity to develop life skills, such as social, behavioral, and moral competencies, that can be generalized to domains beyond the immediate activity. Although context-specific instruments are available to assess developmental outcomes, a measure of life skills transfer would enable evaluation of PYD programs in successfully teaching skills that youth report using in other domains. The purpose of our studies was to develop and validate a measure of perceived life skills transfer, based on data collected with The First Tee, a physical activity-based PYD program. In 3 studies, we conducted a series of steps to provide content and construct validity and internal consistency reliability for the Life Skills Transfer Survey (LSTS), a measure of perceived life skills transfer. Study 1 provided content validity for the LSTS that included 8 life skills and 50 items. Study 2 revealed construct validity (structural validity) through a confirmatory factor analysis and convergent validity by correlating scores on the LSTS with scores on an assessment tool that measures a related construct. Study 3 offered additional construct validity by reassessing youth 1 year later and showing that scores during both time periods were invariant in factor pattern, loadings, and variances and covariances. Studies 2 and 3 demonstrated internal consistency reliability of the LSTS. RESULTS from 3 studies provide evidence of content and construct validity and internal consistency reliability for the LSTS, which can be used in evaluation research with youth development programs.


Iterative design of a helically folded aromatic oligoamide sequence for the selective encapsulation of fructose.
The ab initio design of synthetic molecular receptors for a specific biomolecular guest remains an elusive objective, particularly for targets such as monosaccharides, which have very close structural analogues. Here we report a powerful approach to produce receptors with very high selectivity for specific monosaccharides and, as a demonstration, we develop a foldamer that selectively encapsulates fructose. The approach uses an iterative design process that exploits the modular structure of folded synthetic oligomer sequences in conjunction with molecular modelling and structural characterization to inform subsequent refinements. Starting from a first-principles design taking size, shape and hydrogen-bonding ability into account and using the high predictability of aromatic oligoamide foldamer conformations and their propensity to crystallize, a sequence that binds to β-D-fructopyranose in organic solvents with atomic-scale complementarity was obtained in just a few iterative modifications. This scheme, which mimics the adaptable construction of biopolymers from a limited number of monomer units, provides a general protocol for the development of selective receptors.


Short exposure to actinomycin D induces "reversible" translocation of protein B23 as well as "reversible" inhibition of cell growth and RNA synthesis in HeLa cells.
HeLa cells were grown in medium containing various amounts of actinomycin D for various times. Cellular localization of protein B23 was detected using an immunofluorescence technique. Bright nucleolar fluorescence was observed in untreated cells. A shifting of nucleolar to nuclear fluorescence was observed with increasing doses of actinomycin D and longer incubation times. The degree of translocation of protein B23 from nucleoli to nucleoplasm is dependent on the amount of the drug used and the duration of incubation. Short exposure (0.5 h) of HeLa cells to actinomycin D (0.01-0.25 microgram/ml) induced "reversible" translocation of protein B23, inhibition of cell growth, and RNA synthesis. A majority of cells (greater than 75%) treated with actinomycin D (0.01-0.25 microgram/ml) for 0.5 h still retained bright nucleolar fluorescence. A shifting of nucleolar to nuclear fluorescence as well as inhibition of cell growth and RNA synthesis were observed within 6 h after the removal of the drug. However, at the extended periods (greater than 24 h) after drug removal, RNA synthesis and cell growth resumed at the normal rate, and protein B23 relocated from nucleoplasm to nucleoli. This is in contrast to the results obtained from the experiments using higher doses (1 microgram/ml; 0.5 h) or longer (0.25 microgram/ml; 2 h) exposure of HeLa cells to actinomycin D, which induced irreversible B23 translocation as well as irreversible inhibition of cell growth and RNA synthesis. These results indicated that actinomycin D can be a reversible inhibitor depending on the drug extracellular concentrations and exposure times. Our results also indicated that "B23 translocation" is closely associated with states of cell growth and inhibition of RNA synthesis. "B23 translocation" may therefore be a simple and rapid method for assessing the inhibition of cell growth in response to antitumor therapy.


Rationality, irrationality and escalating behavior in lowest unique bid auctions.
Information technology has revolutionized the traditional structure of markets. The removal of geographical and time constraints has fostered the growth of online auction markets, which now include millions of economic agents worldwide and annual transaction volumes in the billions of dollars. Here, we analyze bid histories of a little studied type of online auctions--lowest unique bid auctions. Similarly to what has been reported for foraging animals searching for scarce food, we find that agents adopt Lévy flight search strategies in their exploration of "bid space". The Lévy regime, which is characterized by a power-law decaying probability distribution of step lengths, holds over nearly three orders of magnitude. We develop a quantitative model for lowest unique bid online auctions that reveals that agents use nearly optimal bidding strategies. However, agents participating in these auctions do not optimize their financial gain. Indeed, as long as there are many auction participants, a rational profit optimizing agent would choose not to participate in these auction markets.


Psychological symptom screening in an adult sickle cell disease clinic and predictors of treatment follow up.
Absence of formal and systematic screening for mood and anxiety disorders among patients with sickle cell disease (SCD) can result in under-recognized psychological problems. This study examined the prevalence of psychological symptoms using a systematic screening process. Patients with SCD completed four self-report screening tools for measurement of depressive and anxiety symptoms, self-efficacy, and pain. The goal was to detect patients with psychological symptoms and identify predictors of follow-up treatment attendance. A total of 336 adult patients (57% female, mean age 33 years) completed validated screening instruments for major depressive disorder and generalized anxiety disorder. Patients recommended for mental health follow-up included higher proportions of women. Patients who accepted the mental health follow up had higher levels of education compared to groups that did not accept nor attend the follow-up appointment. Overall, 34% of patients who endorsed elevated distress scores and were referred for mental health care attended the follow-up appointment. Findings suggest patients with SCD and elevated psychological distress are likely to use mental health treatment resources, which notes this program's success in identifying needs and responding to them. However, further research is needed to understand ways to engage this population in mental health care.


Direct expression of mature bovine adrenodoxin in Escherichia coli.
Site-directed mutagenesis was utilized to enable direct expression of the mature form of bovine adrenodoxin cDNA using the pKK223-3 expression vector in Escherichia coli. Expression was under control of the "tac" promoter and resulted in a direct expression of soluble mature bovine adrenodoxin (greater than 15 mg per liter). Chromatographic behavior of recombinant adrenodoxin did not differ from that reported for mature native adrenodoxin. The purified recombinant protein was identical to native mitochondrial adrenodoxin on the basis of molecular weight, NH2 terminal sequencing and immunoreactivity. E. coli lysates were brown in color, and the purified protein possessed a visible absorbance spectra identical to native bovine adrenodoxin consistent with incorporation of a [2Fe-2S] cluster in vivo. Recombinant bovine adrenodoxin was active in cholesterol side-chain cleavage when reconstituted with adrenodoxin reductase and cytochrome P450scc and exhibited kinetics reported for native bovine adrenodoxin. The presence of the adrenodoxin amino terminal presequence does not appear to be essential for correct folding of mature recombinant adrenodoxin in E. coli. This expression system should prove useful for overexpression of adrenodoxin mutants in future structure/function studies. The approach described herein can potentially be used to directly express the mature form of any protein in bacteria.


Design and evaluation of a novel floating osmotic pump system.
Find a novel delivery system for oral administration of drugs that have absorption window in the upper part of gastrointestinal (GI) track. Dipyridamole was chosen as the model drug. A novel system, which combined the osmotic pump controlled release system and the floating system, was designed; matrix tablets (MT) were prepared for compares. The effects of pH, temperature and hydrodynamic conditions on drug release and the floating behavior of floating osmotic pump system (FOP) were investigated. In vivo evaluation was performed by a three-crossover study in six Beagle dogs relative to the conventional tablet (CT). Cumulative percent input in vivo was compared with that of in vitro release profiles. Floating behavior of FOP, drug releases from FOP and MT were sensitive to pH of dissolution media but not sensitive to temperature; the release of dipyridamole from MT was influenced by stirring rate while drug release from FOP was not. AUC of FOP was larger than MT and CT. The linear correlations between fraction absorbed in vivo and fraction dissolved in vitro was established for FOP-a true zero-order release formula, whereas only a nonlinear correlation was obtained for MT. FOP could be a novel way for the oral administration for drugs like dipyridamole.


Comparison of coil and Kiil dialysers.
To assess the comparative efficiency, safety, and cost of maintenance dialysis, the treatment of 13 patients with a Kiil dialyser (representing 1,477 hospital and 735 home dialyses) was compared with that of 11 patients using a coil dialyser (898 hospital and 396 home dialyses). Kiil and coil dialysers proved equally satisfactory from a medical standpoint and equally acceptable to the patients. The capital costs of home dialysis were considerably reduced without any threat to safety or efficiency. The running costs of coil dialysers approximate to those of Kiil dialysers.


Ecophysiological consequences of alcoholism on human gut microbiota: implications for ethanol-related pathogenesis of colon cancer.
Chronic consumption of excess ethanol increases the risk of colorectal cancer. The pathogenesis of ethanol-related colorectal cancer (ER-CRC) is thought to be partly mediated by gut microbes. Specifically, bacteria in the colon and rectum convert ethanol to acetaldehyde (AcH), which is carcinogenic. However, the effects of chronic ethanol consumption on the human gut microbiome are poorly understood, and the role of gut microbes in the proposed AcH-mediated pathogenesis of ER-CRC remains to be elaborated. Here we analyse and compare the gut microbiota structures of non-alcoholics and alcoholics. The gut microbiotas of alcoholics were diminished in dominant obligate anaerobes (e.g., Bacteroides and Ruminococcus) and enriched in Streptococcus and other minor species. This alteration might be exacerbated by habitual smoking. These observations could at least partly be explained by the susceptibility of obligate anaerobes to reactive oxygen species, which are increased by chronic exposure of the gut mucosa to ethanol. The AcH productivity from ethanol was much lower in the faeces of alcoholic patients than in faeces of non-alcoholic subjects. The faecal phenotype of the alcoholics could be rationalised based on their gut microbiota structures and the ability of gut bacteria to accumulate AcH from ethanol.


Non-Frankia actinomycetes isolated from surface-sterilized roots of Casuarina equisetifolia fix nitrogen.
Based on partial 16S sequences, we previously described a novel group of nonsymbiotic, acetylene reduction activity-positive actinomycetes which were isolated from surface-sterilized roots of Casuarina equisetifolia growing in Mexico. An amplified rRNA restriction analysis confirmed that these actinomycetes are distinct from Frankia, a finding substantiated by a 16S rRNA gene phylogenetic analysis of two of the Mexican isolates. Further support for these actinomycetes being separate from Frankia comes from the very low DNA-DNA homology that was found. Nevertheless, the Mexican isolates may be diazotrophs based not only on their ability to grow in N-free medium and reduce acetylene to ethylene but also on the results from (15)N isotope dilution analysis and the finding that a nifH gene was PCR amplified. A comparison of the nifH sequences from the various isolates showed that they are closely related to nifH from Frankia; the similarity was 84 to 98% depending on the host specificity group. An analysis of complete 16S rRNA gene sequences demonstrated that the two strains analyzed in detail are most closely related to actinobacteria in the Thermomonosporaceae and the Micromonosporaceae.


Glucose Transporters and Virulence in Leishmania mexicana.
Glucose transporters are important for viability and infectivity of the disease-causing amastigote stages of Leishmania mexicana The Δgt1-3 null mutant, in which the 3 clustered glucose transporter genes, GT1, GT2, and GT3, have been deleted, is strongly impaired in growth inside macrophages in vitro We have now demonstrated that this null mutant is also impaired in virulence in the BALB/c murine model of infection and forms lesions considerably more slowly than wild-type parasites. Previously, we established that amplification of the PIFTC3 gene, which encodes an intraflagellar transport protein, both facilitated and accompanied the isolation of the original Δgt1-3 null mutant generated in extracellular insect-stage promastigotes. We have now isolated Δgt1-3 null mutants without coamplification of PIFTC3 These amplicon-negative null mutants are further impaired in growth as promastigotes, compared to the previously described null mutants containing the PIFTC3 amplification. In contrast, the GT3 glucose transporter plays an especially important role in promoting amastigote viability. A line that expresses only the single glucose transporter GT3 grows as well inside macrophages and induces lesions in animals as robustly as do wild-type amastigotes, but lines expressing only the GT1 or GT2 transporters replicate poorly in macrophages. Strikingly, GT3 is restricted largely to the endoplasmic reticulum in intracellular amastigotes. This observation raises the possibility that GT3 may play an important role as an intracellular glucose transporter in the infectious stage of the parasite life cycle.IMPORTANCE Glucose transport plays important roles for in vitro growth of insect-stage promastigotes and especially for viability of intramacrophage mammalian host-stage amastigotes of Leishmania mexicana However, the roles of the three distinct glucose transporters, GT1, GT2, and GT3, in parasite viability inside macrophages and virulence in mice have not been fully explored. Parasite lines expressing GT1 or GT2 alone were strongly impaired in growth inside macrophages, but lines expressing GT3 alone infected macrophages and caused lesions in mice as robustly as wild-type parasites. Notably, GT3 localizes to the endoplasmic reticulum of intracellular amastigotes, suggesting a potential role for salvage of glucose from that organelle for viability of infectious amastigotes. This study establishes the unique role of GT3 for parasite survival inside host macrophages and for robust virulence in infected animals.


Handwritten numeral databases of Indian scripts and multistage recognition of mixed numerals.
This article primarily concerns the problem of isolated handwritten numeral recognition of major Indian scripts. The principal contributions presented here are (a) pioneering development of two databases for handwritten numerals of two most popular Indian scripts, (b) a multistage cascaded recognition scheme using wavelet based multiresolution representations and multilayer perceptron classifiers and (c) application of (b) for the recognition of mixed handwritten numerals of three Indian scripts Devanagari, Bangla and English. The present databases include respectively 22,556 and 23,392 handwritten isolated numeral samples of Devanagari and Bangla collected from real-life situations and these can be made available free of cost to researchers of other academic Institutions. In the proposed scheme, a numeral is subjected to three multilayer perceptron classifiers corresponding to three coarse-to-fine resolution levels in a cascaded manner. If rejection occurred even at the highest resolution, another multilayer perceptron is used as the final attempt to recognize the input numeral by combining the outputs of three classifiers of the previous stages. This scheme has been extended to the situation when the script of a document is not known a priori or the numerals written on a document belong to different scripts. Handwritten numerals in mixed scripts are frequently found in Indian postal mails and table-form documents.


Fast, vacancy-free climb of prismatic dislocation loops in bcc metals.
Vacancy-mediated climb models cannot account for the fast, direct coalescence of dislocation loops seen experimentally. An alternative mechanism, self climb, allows prismatic dislocation loops to move away from their glide surface via pipe diffusion around the loop perimeter, independent of any vacancy atmosphere. Despite the known importance of self climb, theoretical models require a typically unknown activation energy, hindering implementation in materials modeling. Here, extensive molecular statics calculations of pipe diffusion processes around irregular prismatic loops are used to map the energy landscape for self climb in iron and tungsten, finding a simple, material independent energy model after normalizing by the vacancy migration barrier. Kinetic Monte Carlo simulations yield a self climb activation energy of 2 (2.5) times the vacancy migration barrier for 1/2〈111〉 (〈100〉) dislocation loops. Dislocation dynamics simulations allowing self climb and glide show quantitative agreement with transmission electron microscopy observations of climbing prismatic loops in iron and tungsten, confirming that this novel form of vacancy-free climb is many orders of magnitude faster than what is predicted by traditional climb models. Self climb significantly influences the coarsening rate of defect networks, with important implications for post-irradiation annealing.


FAOT-Net: A 1.5-Stage Framework for 3D Pelvic Lymph Node Detection with Online Candidate Tuning.
Accurate and automatic detection of pelvic lymph nodes in computed tomography (CT) scans is critical for diagnosing lymph node metastasis in colorectal cancer, which in turn plays a crucial role in its staging, treatment planning, surgical guidance, and postoperative follow-up of colorectal cancer. However, achieving high detection sensitivity and specificity poses a challenge due to the small and variable sizes of these nodes, as well as the presence of numerous similar signals within the complex pelvic CT image. To tackle these issues, we propose a 3D feature-aware online-tuning network (FAOT-Net) that introduces a novel 1.5-stage structure to seamlessly integrate detection and refinement via our online candidate tuning process and takes advantage of multi-level information through the tailored feature flow. Furthermore, we redesign the anchor fitting and anchor matching strategies to further improve detection performance in a nearly hyperparameter-free manner. Our framework achieves the FROC score of 52.8 and the sensitivity of 91.7% with 16 false positives per scan on the PLNDataset. Code will be available at: github.com/SCUsomebody/FAOT-Net/.


Are Pulley Exercises Initiated 6 Weeks After Rotator Cuff Repair a Safe and Effective Rehabilitative Treatment? A Randomized Controlled Trial.
There are few level 1 or 2 evidence studies that examine rehabilitation after rotator cuff repair. Pulleys have been used in postoperative shoulder rehabilitation with the intention of improving range of motion and developing strength. There is a concern that the use of pulleys in rehabilitation of rotator cuff repairs may contribute to excessive scapular motion (scapular substitution) and potentially inferior outcomes. Rotator cuff repair patients treated with pulley exercises would have increased scapular substitution and inferior patient-determined outcome scores, range of motion, and strength compared with patients treated with an alternative rehabilitation program without pulleys. Randomized controlled trial; Level of evidence, 1. A total of 27 patients who underwent rotator cuff repair were randomized to a rehabilitation group that used pulleys initiated 6 weeks postoperatively, and 26 patients followed a rehabilitation protocol without pulleys. Inclusion criteria were patients undergoing arthroscopic rotator cuff repair. Exclusion criteria were large to massive rotator cuff tears, revision rotator cuff repair, glenohumeral osteoarthritis, adhesive capsulitis, and a symptomatic contralateral shoulder. Outcomes of intervention were patient-determined outcome scores (Western Ontario Rotator Cuff Index [WORC], American Shoulder and Elbow Surgeons [ASES] Shoulder Score, Single Assessment Numeric Evaluation [SANE], Shoulder Activity Level, and Simple Shoulder Test [SST]), range of motion, scapular substitution, and strength. Outcomes were determined at 6, 12, 18, 26, and 52 weeks. A power analysis determined that 22 patients were needed per group to have a power of 0.80, α = 0.05, and effect size of f = 0.5. Both groups had statistically significant improvements in WORC, ASES Shoulder Score, SST, and SANE scores over time after rotator cuff repair (P < .0001). There were no differences between the interventions for WORC (P = .18), ASES Shoulder Score (P = .73), SANE (P = .5), Shoulder Activity Level (P = .39), or SST (P = .36). Both interventions demonstrated improvements in shoulder flexion (P = .002), abduction (P = .0001), external rotation (P = .02), strength (P ≤ .0002), and scapular substitution (P ≤ .07) over time after rotator cuff repair. However, there was no difference in range of motion (P ≥ .26), strength (P ≥ .20), or scapular substitution (P ≥ .17) between interventions. A rotator cuff repair rehabilitation program that uses pulleys does not result in inferior outcomes, as determined by patient-determined outcome scores, measurements of scapular substitution, range of motion, and scaption strength. ClinicalTrials.gov NCT01819909.


Novel CHD7 mutations contributing to the mutation spectrum in patients with CHARGE syndrome.
CHARGE syndrome is an autosomal dominant inherited multiple malformation disorder typically characterized by coloboma, choanal atresia, hypoplastic semicircular canal, cranial nerve defects, cardiovascular malformations and ear abnormalities. Mutations in the chromodomain helicase DNA-binding protein 7 (CHD7) gene are the major cause of CHARGE syndrome. Mutation analysis was performed in 18 patients with firm or tentative clinical diagnosis of CHARGE syndrome. In this study eight mutations distributed across the gene were found. Five novel mutations - one missense (c.2936T > C), one nonsense (c.8093C > A) and three frameshift mutations (c.804_805insAT, c.1757_1770del14, c.1793delA) - were identified. As far as familial data were available these mutations were found to have arisen de novo. Comparison of the clinical features of patients with the same mutation demonstrates that expression of the phenotype is highly variable. The mutation detection rate in this study was 44.4% in patients with a clinically established or suspected diagnosis of CHARGE syndrome.


Staff's Experiences of Preparing and Caring for Children With Cancer and Their Families During the Child's Radiotherapy.
Approximately one-third of children diagnosed with cancer are treated with radiotherapy (RT). Staff experiences of preparing and distracting the children and their families during a child's RT are sparsely described. The aim of this study was to describe staff experiences of preparing and caring for children with cancer and their families during the child's RT. Semistructured interviews with staff were performed at 3 Swedish RT centers. The interviews were analyzed using inductive qualitative content analysis. The analysis revealed 5 categories summarizing the staff members' experiences. These include the following: experiences of various emotions; care for the child and the child's family; commitments before, during, and after RT; organizational issues; and experiences of the intervention and suggestions for improvement. The preparatory intervention facilitated the ability of staff members to conduct their work, although the intervention should be specifically tailored to each child. Meeting children and their families and providing care to both during RT were challenging. The staff strived to provide optimal care for each child and family. Interdisciplinary teamwork and organizational acceptance for the importance of preparation and distraction were essential. A future challenge will be to provide opportunities for all staff involved in the treatment of children with cancer to develop their skills continuously in order to provide high-quality preparation and distraction to all children undergoing RT, regardless of the geographical location of the RT center.


Role of T cell help and endoplasmic reticulum targeting in protective CTL response against influenza virus.
We report on the induction of primary and long-term memory cytotoxic T lymphocyte (CTL) responses against the nucleoprotein of the influenza virus A/PR8/34 in mice immunized with plasmid DNA targeted to B lymphocytes in the spleen. We found that the magnitude of the CTL response and the size of the pool of memory CTL was greater when the CTL response was induced in presence of T cell help. Interestingly, immunization with a signal sequence-competent transgene was markedly superior to immunization with a transgene lacking the endoplasmic reticulum (ER) targeting sequence, in inducing CTL. We also found a correlation between in vivo protection from lethal virus challenge and (1) the availability of T cell help and (2) ER targeting. Immunization of dendritic cell-deficient mice suggests that B lymphocytes function as antigen-presenting cells in this model of immunization. Collectively, the results suggest that somatic transgene immunization is a conceptually new approach to induce effective anti-viral CTL responses and to assess the parameters critical for long-lasting and protective CTL responses in vivo.


Antibacterial and antifungal properties of acetonic extract of Feijoa sellowiana fruits and its effect on Helicobacter pylori growth.
Feijoa sellowiana Berg. fruit, widely used for human consumption, is well appreciated for its good nutritional characteristics and for its pleasant flavor and aroma. In a prior study we showed that the acetonic extract of F. sellowiana fruit exerts a potent antibacterial activity against some Gram-positive and Gram-negative bacterial strains. To confirm and deepen the study on antimicrobial activities of the fruit, in this article we analyze the same extract for its effectiveness against different kind of bacteria both as American Type Culture Collection standard strains and clinically isolates, the Gram-negative Helicobacter pylori, and three fungal strains (one human pathogen and two phytopathogens). Our aim was also to isolate and define the active component(s) involved in the antibacterial, antifungal, and anti-H. pylori activity of the extract through activity-guided fractionation procedures. This allowed us to identify flavone as the active compound of F. sellowiana fruit. Flavone showed a high antibacterial activity against the nine standard bacterial strains tested and the matched clinically isolated bacterial strains and was significantly more active against H. pylori than metronidazole. Pseudomonas aeruginosa, Proteus mirabilis, and Proteus vulgaris from both standard and clinical sources showed a very high sensitivity to flavone (minimal inhibitory concentration [MIC] = 1.95 microg/mL for standard strains and 3.9 microg/mL for clinical isolates of the three bacteria), showing also good minimal bactericidal concentration values. Among fungi tested, Rhyzoctonia solani was the most sensitive strain to the action of flavone (MIC = 62.5 microg/mL).


Diruthenium-polyyn-diyl-diruthenium wires: electronic coupling in the long distance regime.
Reported herein is a series of Ru2(Xap)4 capped polyyn-diyl compounds, where Xap is either 2-anilinopyridinate (ap) or its aniline substituted derivatives. Symmetric [Ru2(Xap)4](μ-C4k)[Ru2(Xap)4] (compounds 4ka (X = 3-isobutoxy) and 4kc (X = 3,5-dimethoxy) with k = 2, 3, 4, and 5) was obtained from the Glaser coupling reaction of Ru2(Xap)4(C2kH). Unsymmetric [Ru2(Xap)4](μ-C(4k+2))[Ru2(ap)4] (compounds 4k+2b with k = 2, 3, and 4) were prepared from the Glaser coupling reaction between Ru2(Xap)4(C(2k+2)H) and Ru2(ap)4(C2kH). X-ray diffraction study of compound 12c revealed both the sigmoidal topology of the polyyn-diyl bridge and the fine structural detail about the Ru2 cores. Cyclic and differential pulse voltammetric (CV and DPV) measurements and spectroelectrochemical studies revealed that (i) the reduced monoanions [Ru2-C2m-Ru2](-1) (m = 4-8) belong to the Robin-Day class II mixed valent ions and (ii) the electronic coupling between Ru2 termini depends on the length of the polyyn-diyl bridge with an attenuation constant (γ) between 0.12 and 0.15 Å(-1). In addition, spin-unrestricted DFT calculations provide insight about the nature of orbitals that mediate the long distance electronic coupling.


Atrial natriuretic peptide: physiological release associated with natriuresis during water immersion in man.
Thermoneutral water immersion produces a physiological increase of thoracic blood volume, raises central venous pressure and increases urinary sodium excretion by a hitherto ill-understood mechanism. We have investigated whether this enhanced sodium excretion could be mediated by the recently discovered natriuretic factor, atrial natriuretic peptide (ANP). During water immersion there was a highly significant (P less than 0.001) twofold increase of the mean plasma ANP concentration and a doubling of the mean urinary sodium excretion. Both were unchanged during the control experiments. These results are consistent with the hypotheses that ANP is released into plasma in response to central blood volume expansion and that it functions as a natriuretic hormone in normal man under physiological conditions.


Differential Expression of Lipoprotein-Associated Phospholipase A2 in Monocyte Subsets: Impact of Uremia and Atherosclerosis.
Monocytic products, such as lipoprotein-associated phospholipase A2 (Lp-PLA2), may participate in the development of atherosclerosis. Heterogeneity of monocytes is widely acknowledged. Classical, intermediate, and non-classical subsets can be discerned. Recently, an inflammatory, pro-atherogenic monocyte population could be identified in hemodialysis patients. In this study, we investigated the expression of Lp-PLA2 on leucocytes and different monocyte subpopulations and their possible role in uremia, inflammation, and atherosclerosis. Chronic kidney disease stage 5-D (CKD5-D; n = 57), healthy control subjects with hs-C-reactive protein (CRP) levels ≤1 mg/L (CO-N, n = 22) and a control group with inflammatory activation (CRP levels >1 mg/L, CO-I, n = 29) were enrolled in this cross-sectional observation. The CKD5-D cohort was dichotomized into patients with (A+) and without subclinical atherosclerosis (A-) by carotid artery ultrasound measurement. Lp-PLA2 activity was determined in plasma samples, Lp-PLA2 mRNA expression analysis in leucocytes, and sorted monocyte subsets. Effects of Lp-PLA2 overexpression were studied in classical vs. intermediate and non-classical subsets. The classical monocytes expressed the highest Lp-PLA2 mRNA levels as compared to other subpopulations. CKD5-D patients revealed significantly higher Lp-PLA2 transcripts, as well as higher Lp-PLA2 plasma activity as compared to healthy and "inflammatory" controls. In vitro data confirmed that uremia significantly contributes to Lp-PLA2 mRNA upregulation. Non-classical monocytes of A+ patients revealed significant higher Lp-PLA2 mRNA compared to A-. Uremic environment but not inflammation per se increases plasma Lp-PLA2 activity and upregulates monocytic Lp-PLA2 mRNA expression. The highest Lp-PLA2 levels were found in the classical and not in the inflammatory subsets. Atherosclerosis also contributes to a subset-specific increase in Lp-PLA2 mRNA expression.


Improving the accuracy of acetabular component orientation: avoiding malposition.
Improper acetabular component orientation negatively affects the outcome of total hip arthroplasty through increasing dislocation rates, component impingement, bearing surface wear, and the number of revision surgeries. Leg length, hip biomechanics, pelvic osteolysis, and acetabular component migration are also affected by malposition. With conventional techniques, numerous variables, such as patient size, deformity and/or position, and decreased visualization, contribute to inter- and intrasurgeon acetabular component variability during surgery regardless of surgeon experience and practice volume. New acetabular component implantation techniques, such as patient-specific morphology, that incorporate anatomic landmarks may provide more accurate and individualized target zones. These techniques, coupled with the use of quantitative technology such as computer-aided navigation, may improve the precision of acetabular component placement.


HkRP3 is a microtubule-binding protein regulating lytic granule clustering and NK cell killing.
NK cells provide host defense by killing viral-infected and cancerous cells through the secretion of preformed lytic granules. Polarization of the lytic granules toward the target cell is dependent on an intact microtubule (MT) network as well as MT motors. We have recently shown that DOCK8, a gene mutated in a primary immunodeficiency syndrome, is involved in NK cell killing in part through its effects on MT organizing center (MTOC) polarization. In this study, we identified Hook-related protein 3 (HkRP3) as a novel DOCK8- and MT-binding protein. We further show that HkRP3 is present in lytic granule fractions and interacts with the dynein motor complex and MTs. Significantly, depletion of HkPR3 impaired NK cell cytotoxicity, which could be attributed to a defect in not only MTOC polarity, but also impaired clustering of lytic granules around the MTOC. Our results demonstrate an important role for HkRP3 in regulating the clustering of lytic granules and MTOC repositioning during the development of NK cell-mediated killing.


The neural signature of escalating frustration in humans.
Mammalian studies show that frustration is experienced when goal-directed activity is blocked. Despite frustration's strongly negative role in health, aggression and social relationships, the neural mechanisms are not well understood. To address this we developed a task in which participants were blocked from obtaining a reward, an established method of producing frustration. Levels of experienced frustration were parametrically varied by manipulating the participants' motivation to obtain the reward prior to blocking. This was achieved by varying the participants' proximity to a reward and the amount of effort expended in attempting to acquire it. In experiment 1, we confirmed that proximity and expended effort independently enhanced participants' self-reported desire to obtain the reward, and their self-reported frustration and response vigor (key-press force) following blocking. In experiment 2, we used functional magnetic resonance imaging (fMRI) to show that both proximity and expended effort modulated brain responses to blocked reward in regions implicated in animal models of reactive aggression, including the amygdala, midbrain periaqueductal grey (PAG), insula and prefrontal cortex. Our findings suggest that frustration may serve an energizing function, translating unfulfilled motivation into aggressive-like surges via a cortical, amygdala and PAG network.


Treatment Patterns Among Patients With Advanced Prostate Cancer in Brazil: An Analysis of a Private Healthcare System Database.
With the ongoing expansion of life-prolonging therapies approved to treat advanced prostate cancer, there is currently an unmet need to better understand real-world treatment patterns and identify optimal treatment sequencing for men with metastatic castration-resistant prostate cancer (mCRPC). In this retrospective, observational cohort analysis, patients with confirmed mCRPC were identified in the Auditron claims database and used to describe mCRPC treatment patterns and trends in the Brazilian private healthcare system from 2014 to 2019. Demographics and clinical characteristics, prostate cancer stage at diagnosis, and type and number of treatment lines were evaluated. The primary endpoint was identification of the drugs used in first-line therapies in mCRPC, and the secondary endpoint included a description of sequential lines of therapy (second and third lines) in mCRPC. A total of 168 electronic patient records were reviewed. Docetaxel was the most frequently used first-line treatment (35.7%), followed by abiraterone (33.3%) and enzalutamide (13.1%). Docetaxel, abiraterone, and enzalutamide also accounted for 34.6%, 28.0%, and 15.0%, respectively, of second-line therapies. In third-line therapies, cabazitaxel (26.1%), enzalutamide (23.9%), docetaxel (15.2%), and abiraterone (15.2%) were most commonly prescribed. Irrespective of stage at diagnosis, treatment patterns were similar once the disease progressed to the metastatic castration-resistance stage. Docetaxel was the most frequently utilized therapy for mCRPC treatment, followed by abiraterone and enzalutamide. Although the current analyses provide real-world insights into treatment patterns for patients with mCRPC in Brazil, additional real-world data are needed to further validate and expand on these findings.


Cryopreservation of spin-dried mammalian cells.
This study reports an alternative approach to achieve vitrification where cells are pre-desiccated prior to cooling to cryogenic temperatures for storage. Chinese Hamster Ovary (CHO) cells suspended in a trehalose solution were rapidly and uniformly desiccated to a low moisture content (<0.12 g of water per g of dry weight) using a spin-drying technique. Trehalose was also introduced into the cells using a high-capacity trehalose transporter (TRET1). Fourier Transform Infrared Spectroscopy (FTIR) was used to examine the uniformity of water concentration distribution in the spin-dried samples. 62% of the cells were shown to survive spin-drying in the presence of trehalose following immediate rehydration. The spin-dried samples were stored in liquid nitrogen (LN(2)) at a vitrified state. It was shown that following re-warming to room temperature and re-hydration with a fully complemented cell culture medium, 51% of the spin-dried and vitrified cells survived and demonstrated normal growth characteristics. Spin-drying is a novel strategy that can be used to improve cryopreservation outcome by promoting rapid vitrification.


Variation in the wood anatomical structure of Gmelina arborea (Verbenaceae) trees at different ecological conditions in Costa Rica.
The tree Gmelina arborea has been widely introduced in Costa Rica for commercial purposes. This new conditions for melina cause variations on anatomy in secondary xylem of the trees growing in plantations. The objective of the present research was to determine the variation in the anatomy of xylem caused by the ecological conduction variation. Dimensions of fiber, axial parenchyma percentage of cross sections, parameters of vessels and the ray were measured. The results showed that some anatomical characteristics remained stable despite variations of ecological conditions, especially radial parenchyma and anatomical features which were less affected by the altitude. On the other hand, the vessels, axial parenchyma and fiber were less stable because they were affected significantly by the longitude, latitude, altitude and precipitation. Latitude significantly affected vessel percentage, length and diameter of the fiber and lumen. Longitude affected vessel percentage and fiber diameter. Altitude had a significant correlation with the amount of cells at ray height. Annual average precipitation affected vessel percentage and diameter, not only of the fiber, but also of the lumen. These results suggest that the new growth conditions of G. arborea trees in Costa Rica have produced an anatomic adaptation.


Cockroach allergy in a group of Turkish children with respiratory allergies.
Exposure to cockroach may lead to exacerbations of bronchial asthma and/or allergic rhinitis in sensitized patients. Although there is a widespread belief that cockroach allergy is a common problem in patients with respiratory allergies, little is known in Turkish children. In order to investigate the prevalence and characteristics of cockroach allergy in respiratory allergic children, we performed a study in newly referred children with respiratory allergies. All patients underwent questionnaire-interview and skin prick tested with common inhalant allergens in addition to two cockroach allergens: Blatella germanica (Bg) and Periplaneta americana (Pa). A subgroup of patients was also serologically investigated for specific IgE against Bg and house dust mite. Three hundred- and thirty-seven children aged 2-16 years were recruited for the study and 77.7% of these were atopic, with the most common indoor and outdoor allergens of house dust mite (47.5%) and grass pollens (45.1%), respectively. According to the prick test results, allergies to Bg and Pa were 11.9% and 7.4%, respectively, and there was a weak correlation between size of the prick test and specific IgE levels for Bg allergen. Almost 30% of the cockroach-sensitive patients were allergic to both cockroach antigens. Seventy percent of cockroach-sensitive patients were also sensitive to house dust mite, and only 1% were monosensitive. Dwellings in the Middle Anatolia and Black Sea regions were less commonly infested by cockroach compared to the dwellings in other regions. In conclusion, our preliminary study showed that cockroach sensitization is common among children with respiratory allergies irrespective of infestation history, suggesting that addition of cockroach allergen to the routine allergy screening panel is critical.


Drive for thinness and fear of fat: separate yet related constructs?
The terms drive for thinness and fear of fat are consistently used in the literature to describe eating disorders in their clinical and sub-clinical forms, yet distinct definitions of each are not provided. Although similar on first examination, these terms may represent two distinct constructs in the motivational factors of persons at risk for developing eating disorders. The following article presents the notion that drive for thinness and fear of fat are separate yet related constructs, perhaps representing approach and avoidance mechanisms in persons at risk. The implications for counseling and practice and directions for further research are provided.


Electrostatic potential wells for on-demand drop manipulation in microchannels.
Precise control and manipulation of individual drops are crucial in many lab-on-a-chip applications. We present a novel hybrid concept for channel-based discrete microfluidics with integrated electrowetting functionality by incorporating co-planar electrodes (separated by a narrow gap) in one of the microchannel walls. By combining the high throughput of channel-based microfluidics with the individual drop control achieved using electrical actuation, we acquire the strengths of both worlds. The tunable strength of the electrostatic forces enables a wide range of drop manipulations, such as on-demand trapping and release, guiding, and sorting of drops in the microchannel. In each of these scenarios, the retaining electrostatic force competes with the hydrodynamic drag force. The conditions for trapping can be predicted using a simple model that balances these forces.


A case report study of 72 consecutive Emdogain-treated intrabony periodontal defects: clinical and radiographic findings after 1 year.
This case report study examined the effect of adjunctive use of Emdogain in the treatment of intrabony periodontal defects. Seventy-two consecutively treated defects in sixty-one patients were included. Efficacy of treatment was evaluated at 12 months by assessment of probing depth reduction, probing attachment level gain, and radiographic bone gain from standardized radiographs. Initial pocket depth averaged 8.3 mm, and the mean probing attachment level was 10.0 mm. The mean radiographic defect depth was 5.3 mm. At 12 months, mean pocket depth reduction was 4.7 mm and mean probing attachment level gain was 4.2 mm. Radiographic bone level gain averaged 3.1 mm and defect fill averaged 70%. The only variables significantly affecting radiographic bone gain were bleeding on probing and smoking. It was concluded that Emdogain treatment of one- and two-walled intrabony defects in a periodontal practice will result in a clinically significant gain of probing attachment level and radiographic bone that is similar to that reported in controlled clinical trials.


River regulation and recruitment in a protracted-spawning riverine fish.
We present and test an extension of the "match/mismatch" hypothesis that attempts to explain the persistence, under conditions of flow alteration, of small, short-lived, native, riverine, fish species. The premise is that flow alteration typically changes environmental conditions, such as temperature and prey abundance, which may affect survival during the larval period of fishes. This "window-of-opportunity hypothesis" states that, if optimal conditions for recruitment vary temporally within a year, the probability that a proportion of the larvae of protracted-spawning species will encounter a period of optimal conditions is greater than for larvae with only a brief spawning period, and so the former will have a recruitment advantage. We determined whether all hatching events contributed equally to juvenile recruitment of the protracted-spawning Australian smelt (Retropinna semoni) during one breeding season in three pairs of heavily regulated and largely free-flowing unregulated rivers in the Murray-Darling Basin, Australia, and related patterns in the hatch dates of recruits to temperature or prey biomass for one pair. For all rivers, heavily regulated or not, recruits present at the end of the breeding season most commonly hatched in the latter part of the breeding season. Mortality of those fish hatched in the first part of the season likely explains this trend. Furthermore, while hatching times were similar for all rivers, each river showed a distinct pattern of hatching and recruitment, which may relate to the temperature range within which epigenetic processes are aligned. Patterns of zooplankton biomass differed between the largely free-flowing ovens and regulated Goulburn rivers and likely had different sources: within the channel and within the storage lake, respectively. For the Ovens River, recruits hatched subsequent to the period when the first significant increase in zooplankton biomass occurred. We hypothesize that temperature may largely influence the "window" during which recruitment can take place but that prey density, responding to river-specific interactions between temperature and discharge, plays a role in the timing and magnitude of recruitment of Australian smelt. We conclude that the match/mismatch hypothesis may be applicable to rivers, that the window-of-opportunity hypothesis has some currency and deserves further investigation, and that river regulation may have significant impacts on fish recruitment.


Correlation of Alzheimer disease neuropathologic changes with cognitive status: a review of the literature.
Clinicopathologic correlation studies are critically important for the field of Alzheimer disease (AD) research. Studies on human subjects with autopsy confirmation entail numerous potential biases that affect both their general applicability and the validity of the correlations. Many sources of data variability can weaken the apparent correlation between cognitive status and AD neuropathologic changes. Indeed, most persons in advanced old age have significant non-AD brain lesions that may alter cognition independently of AD. Worldwide research efforts have evaluated thousands of human subjects to assess the causes of cognitive impairment in the elderly, and these studies have been interpreted in different ways. We review the literature focusing on the correlation of AD neuropathologic changes (i.e. β-amyloid plaques and neurofibrillary tangles) with cognitive impairment. We discuss the various patterns of brain changes that have been observed in elderly individuals to provide a perspective for understanding AD clinicopathologic correlation and conclude that evidence from many independent research centers strongly supports the existence of a specific disease, as defined by the presence of Aβ plaques and neurofibrillary tangles. Although Aβ plaques may play a key role in AD pathogenesis, the severity of cognitive impairment correlates best with the burden of neocortical neurofibrillary tangles.


Relationship Between Glycated Hemoglobin and Vibration Perception Threshold in Diabetic Peripheral Neuropathy.
Foot ulcers, infections, and deformity are some of the major sources of mortality and morbidity among the diabetic population. The relationship between glycated hemoglobin (HbA1c) and diabetic peripheral neuropathy (DPN) has been well established. There is a dearth of literature on the relationship between vibration perception threshold (VPT) and HbA1c values. So, the objective of the study was to determine the strength of linear relationship between HbA1c levels and vibration perception threshold in DPN. This cross-sectional study was conducted at Kasturba Hospital, Manipal, and diabetic foot screening camps held at various parts of Udupi district. Ethical approval was obtained from the Institutional Ethics Committee, Kasturba Hospital, Manipal (IEC:281/2017). A total of 534 participants ranging from 30 to 70 years of age and were diagnosed with type 2 diabetes mellitus on medications were included in the study. Neuropathy assessment consisting of monofilament and vibration perception threshold was done using Neurotouch beta version (Yostra Labs, Bengaluru, India). HbA1c measurement was done using turbidimetric inhibition immunoassay technique (Roche Diagnostics, Mannheim, Germany). Pearson correlation coefficient showed a moderate to good correlation between HbA1c and VPT (r = .0.753, P < .001). Linear regression result has shown a significant relationship of VPT with HbA1c (4.033 [95% confidence interval = 3.67-4.39]). The present study has concluded that there is strong relationship between HbA1c values and VPT and could be a predictor for complications in the foot following DPN.


Retentive strength of fiber-reinforced composite posts with composite resin cores: Effect of remaining coronal structure and root canal dentin conditioning protocols.
The prognosis of a fixed dental prosthesis cemented to endodontically treated teeth is primarily determined by the presence of a ferrule on the tooth. Adhesion of the post in the root canal, conditioning methods for the canal and the amount of coronal structure could also be decisive on survival of reconstructions cemented on endodontically treated teeth. The purpose of this in vitro study was to test the effect of remaining coronal structure on the retention of airborne-particle abraded fiber-reinforced composite resin posts built up with composite resin cores after the treatment of root canal dentin with different conditioning protocols. One hundred and fifty extracted human teeth with single root canal space were endodontically treated and divided into 3 groups as follows: group CEJ: the teeth were sectioned at the level of cementoenamel junction (CEJ); group CEJ1: the teeth were sectioned 1 mm above the CEJ; group CEJ2: the teeth were sectioned 2 mm above the CEJ. Each group was further divided into 5 subgroups (n=10 per group) according to the root canal treatments as follows: group C: no conditioning (control); group PH: conditioning with 37% phosphoric acid gel for 15 seconds; group E: conditioning with 17% ethylenediaminetetraacetic acid (EDTA) for 60 seconds; group CHX: conditioning with 2% chlorhexidine (CHX) for 60 seconds; group Q: conditioning with combination of 2% CHX with 17% EDTA and a surfactant solution for 60 seconds. Glass fiber-reinforced composite resin posts were airborne-particle abraded and luted to the root canal dentin with a self-adhesive resin cement (RelyX Unicem). The retentive force was tested by applying a tensile load parallel to the long axis of these posts at a crosshead speed of 2 mm/min. Two-way ANOVA and the Tukey HSD post hoc test were used to analyze the data. The highest retention (N) was obtained with the CHX-EDTA conditioned group (374.7 ±29.8) followed by 17% EDTA (367.9 ±33.3) conditioning when 2 mm remaining coronal structure was available. Conditioning with the CHX-EDTA showed comparable retention values to 17% EDTA conditioned groups when 0 or 1 mm coronal structure was present that differed significantly compared to 37% PH and 2% CHX conditioning (P<.05). Conditioning root canal either with CHX-EDTA or 17% EDTA delivered superior retention values for fiber-reinforced composite resin posts with composite resin cores that were luted with self-adhesive resin cement to endodontically treated teeth with 2 mm remaining coronal structure.


Expression profiling of archival tumors for long-term health studies.
More than 20 million archival tissue samples are stored annually in the United States as formalin-fixed, paraffin-embedded (FFPE) blocks, but RNA degradation during fixation and storage has prevented their use for transcriptional profiling. New and highly sensitive assays for whole-transcriptome microarray analysis of FFPE tissues are now available, but resulting data include noise and variability for which previous expression array methods are inadequate. We present the two largest whole-genome expression studies from FFPE tissues to date, comprising 1,003 colorectal cancer (CRC) and 168 breast cancer samples, combined with a meta-analysis of 14 new and published FFPE microarray datasets. We develop and validate quality control (QC) methods through technical replication, independent samples, comparison to results from fresh-frozen tissue, and recovery of expected associations between gene expression and protein abundance. Archival tissues from large, multicenter studies showed a much wider range of transcriptional data quality relative to smaller or frozen tissue studies and required stringent QC for subsequent analysis. We developed novel methods for such QC of archival tissue expression profiles based on sample dynamic range and per-study median profile. This enabled validated identification of gene signatures of microsatellite instability and additional features of CRC, and improved recovery of associations between gene expression and protein abundance of MLH1, FASN, CDX2, MGMT, and SIRT1 in CRC tumors. These methods for large-scale QC of FFPE expression profiles enable study of the cancer transcriptome in relation to extensive clinicopathological information, tumor molecular biomarkers, and long-term lifestyle and outcome data.


Which comes first? Psychogenic dizziness versus otogenic anxiety.
OBJECTIVE To investigate the hypotheses that physical neurotologic conditions may trigger anxiety disorders (otogenic pattern of illness), that psychiatric disorders may produce dizziness (psychogenic pattern), and that risk factors for these syndromes may be identified. STUDY DESIGN Retrospective review of all patients (N = 132) treated at a tertiary care balance center from 1998 to 2002 for psychogenic dizziness with or without physical neurotologic illnesses. METHODS All patients underwent comprehensive neurotologic and psychiatric evaluations with attention to the longitudinal course of symptoms and risk factors for psychopathology. Patients were grouped according to the condition first causing dizziness. Risk factors were compared across groups. RESULTS Three equally prevalent patterns of illness were found: anxiety disorders as the sole cause of dizziness (33% of cases), neurotologic conditions exacerbating preexisting psychiatric disorders (34%), and neurotologic conditions triggering new anxiety or depressive disorders (33%). Panic disorder and agoraphobia were significantly more prevalent than less severe phobias in the first two groups, whereas the opposite pattern existed in the third group (P <.0001). More patients in the first two groups had risk factors for anxiety disorders (P <.05). Depression was not a primary cause of dizziness in any patient. Vestibular neuronitis, benign paroxysmal positional vertigo, and migraine were the most common neurotologic conditions. CONCLUSIONS These data support the hypothesis that physical neurotologic conditions may trigger psychopathology as often as primary anxiety disorders cause dizziness. A third pattern appears to be equally common wherein physical neurotologic conditions exacerbate preexisting psychiatric illnesses. Individuals at risk for anxiety disorders may be more likely to have primary psychopathology.


Tea consumption and prostate cancer: an updated meta-analysis.
Tea is supposed to have chemopreventive effect against various cancers. However, the protective role of tea in prostate cancer is still controversial. The aim of this study is to elucidate the association between tea consumption and prostate cancer risk by meta-analysis. A total of 21 published articles were retrieved via both computerized searches and review of references. Estimates of OR/RR for highest versus non/lowest tea consumption levels were pooled on the basis of random effect model or fixed effect model as appropriate. Stratified analyses on tea type, population and study design were also conducted. No statistical significance was detected between tea consumption and prostate cancer risk in meta-analysis of all included studies (odds ratio (OR) = 0.86, 95% CI (0.69-1.04)). Furthermore, stratified analyses on population (Asian, OR = 0.81, 95% CI (0.55-1.08); non-Asian, OR = 0.89, 95% CI (0.72-1.07)) and tea type (green tea, OR = 0.79, 95% CI (0.43-1.14); black tea, OR = 0.88, 95% CI (0.73-1.02)) also yielded non-significant association. Only the case-control study subgroup demonstrated a borderline protective effect for tea consumption against prostate cancer (OR = 0.77, 95% CI (0.55-0.98)). Our analyses did not support the conclusion that tea consumption could reduce prostate cancer risk. Further epidemiology studies are needed.


Cancer-ID: Toward Identification of Cancer by Tumor-Derived Extracellular Vesicles in Blood.
Extracellular vesicles (EVs) have great potential as biomarkers since their composition and concentration in biofluids are disease state dependent and their cargo can contain disease-related information. Large tumor-derived EVs (tdEVs, >1 μm) in blood from cancer patients are associated with poor outcome, and changes in their number can be used to monitor therapy effectiveness. Whereas, small tumor-derived EVs (<1 μm) are likely to outnumber their larger counterparts, thereby offering better statistical significance, identification and quantification of small tdEVs are more challenging. In the blood of cancer patients, a subpopulation of EVs originate from tumor cells, but these EVs are outnumbered by non-EV particles and EVs from other origin. In the Dutch NWO Perspectief Cancer-ID program, we developed and evaluated detection and characterization techniques to distinguish EVs from non-EV particles and other EVs. Despite low signal amplitudes, we identified characteristics of these small tdEVs that may enable the enumeration of small tdEVs and extract relevant information. The insights obtained from Cancer-ID can help to explore the full potential of tdEVs in the clinic.


Constructing biological pathway models with hybrid functional Petri nets.
In many research projects on modeling and analyzing biological pathways, the Petri net has been recognized as a promising method for representing biological pathways. From the pioneering works by Reddy et al., 1993, and Hofestädt, 1994, that model metabolic pathways by traditional Petri net, several enhanced Petri nets such as colored Petri net, stochastic Petri net, and hybrid Petri net have been used for modeling biological phenomena. Recently, Matsuno et al., 2003b, introduced the hybrid functional Petri net (HFPN) in order to give a more intuitive and natural modeling method for biological pathways than these existing Petri nets. Although the paper demonstrates the effectiveness of HFPN with two examples of gene regulation mechanism for circadian rhythms and apoptosis signaling pathway, there has been no detailed explanation about the method of HFPN construction for these examples. The purpose of this paper is to describe method to construct biological pathways with the HFPN step-by-step. The method is demonstrated by the well-known glycolytic pathway controlled by the lac operon gene regulatory mechanism.


Functional domains of SIR4, a gene required for position effect regulation in Saccharomyces cerevisiae.
The product of the Saccharomyces cerevisiae SIR4 gene, in conjunction with at least three other gene products, prevents expression of mating-type genes resident at loci at either end of chromosome III, but not of the same genes resident at the MAT locus in the middle of the chromosome. To address the mechanism of this novel position effect regulation, we have conducted a structural and genetic analysis of the SIR4 gene. We have determined the nucleotide sequence of the gene and found that it encodes a lysine-rich, serine-rich protein of 152 kilodaltons. Expression of the carboxy half of the protein complements a chromosomal nonsense mutation of sir4 but not a complete deletion of the gene. These results suggest that SIR4 protein activity resides in two portions of the molecule, but that these domains need not be covalently linked to execute their biological function. We also found that high-level expression of the carboxy domain of the protein yields dominant derepression of the silent loci. This anti-Sir activity can be reversed by increased expression of the SIR3 gene, whose product is normally also required for maintaining repression of the silent loci. These results are consistent with the hypothesis that SIR3 and SIR4 proteins physically associate to form a multicomponent complex required for repression of the silent mating-type loci.


Currarino triad: surgical management and follow-up results of four [correction of three] cases.
The Currarino syndrome is a rare triad that is a combination of a presacral mass, a congenital sacral bony abnormality and an anorectal malformation. We present 4 children with complete Currarino triad who were diagnosed using MRI. Our aim was to report the neurosurgical management of Currarino syndrome in children. All of the patients had chronic constipation and pain in the lumbosacral region. In the plain radiograph, 3 patients had a sacral scimitar-shaped bony abnormality, and 1 patient had total sacral agenesis. There was a narrow anal canal or narrow ventrally displaced anus in all patients. Their anorectal malformations were characterized as anal stenoses (4 patients), associated with Hirschsprung's disease in 2 cases. In 3 patients, MRI showed tethered cord syndrome in addition to the presacral mass. There was hydrocephalus in 1 patient. Anal stenosis was treated by anal dilatation. In 2 patients, rectal biopsy and temporary colostomy (2 patients) had been performed previously due to Hirschsprung's disease. We performed a posterior procedure via lumbar and sacral partial laminectomy-laminoplasty and transdural ligation of the neck of the meningocele for anterior sacral meningoceles, or alternatively, tumor excision for other types of presacral lesions. Histopathologically, 3 were cases of anterior sacral meningoceles and 1 was a teratoma. One of them also had a spinal abscess. He required reoperation (twice) and appeared at the time to have improved with medical therapy. All patients improved and stabilized. There were no additional neurological deficits and no recurrence of the presacral mass over the follow-up period (6 years, on average). The family pedigree did not reveal any familial transmission pattern. In cases of Currarino triad, MRI can allow the characterization of the presacral masses. If it is an anterior sacral meningocele or a solid tumor without severe anorectal malformation, it can be managed with posterior lumbar and sacral procedures. Such approaches are performed easily by transdural ligation of the neck of the anterior sacral meningocele or through tumor excision.