Workshop on Measurement Needs for Local-Structure Determination in Inorganic Materials.
The functional responses (e.g., dielectric, magnetic, catalytic, etc.) of many industrially-relevant materials are controlled by their local structure-a term that refers to the atomic arrangements on a scale ranging from atomic (sub-nanometer) to several nanometers. Thus, accurate knowledge of local structure is central to understanding the properties of nanostructured materials, thereby placing the problem of determining atomic positions on the nanoscale-the so-called "nanostructure problem"-at the center of modern materials development. Today, multiple experimental techniques exist for probing local atomic arrangements; nonetheless, finding accurate comprehensive, and robust structural solutions for the nanostructured materials still remains a formidable challenge because any one of these methods yields only a partial view of the local structure. The primary goal of this 2-day NIST-sponsored workshop was to bring together experts in the key experimental and theoretical areas relevant to local-structure determination to devise a strategy for the collaborative effort required to develop a comprehensive measurement solution on the local scale. The participants unanimously agreed that solving the nanostructure problem-an ultimate frontier in materials characterization-necessitates a coordinated interdisciplinary effort that transcends the existing capabilities of any single institution, including national laboratories, centers, and user facilities. The discussions converged on an institute dedicated to local structure determination as the most viable organizational platform for successfully addressing the nanostructure problem. The proposed "institute" would provide an intellectual infrastructure for local structure determination by (1) developing and maintaining relevant computer software integrated in an open-source global optimization framework (Fig. 2), (2) connecting industrial and academic users with experts in measurement techniques, (3) developing and maintaining pertinent databases, and (4) providing necessary education and training.


Social and proximate determinants of sexual activity in rural Nigeria.
This study analyses the social, physiological and motivational determinants of sexual activity for 644 married women in rural areas of Osun State, Nigeria. The data come from the Fertility Awareness and Pregnancy Avoidance study conducted in 1993-94. Sexual activity is measured by three continuous variables: weekly frequency of (1) total sexual activity, (2) coital-only sex and (3) both coital/non-coital sex. Analyses of variance were performed to test the difference in group means between the predictor variables and measures of sexual activity. Ordinary least squares regression analyses were then performed for the three dependent variables. Two models are used: the first contains only those variables associated with an individual's demographic, social and economic status, and the second adds predictor variables associated with motivational and physiological factors. Results show that while many socioeconomic variables by themselves are significantly associated with sexual activity measures, the addition of physiological and motivational variables weakens their effects and lessens their original statistical significance. Additionally, the socioeconomic and demographic factors associated with frequency of sexual activity are not necessarily the same ones significantly associated with coital-only or combined coital/non-coital sex. While other studies have tended to focus either on socioeconomic or physiological factors, the joint examination of both types of influences seems to indicate that the latter have more proximate effects on the frequency of both coital and non-coital sexual activity of married women in this population. Contraceptive protection is shown to be highly positively associated with all three of the sexual activity measures, empirically confirming the important relationship between contraception and both coital and non-coital forms of sexual activity.


Bystander effects in tumor-free and tumor-bearing rat brains following irradiation by synchrotron X-rays.
Synchrotron microbeam radiation therapy (MRT) is a radiosurgery concept in the preclinical stage, developed mainly for brain tumor treatment. Experimental studies suggest that with MRT a better therapeutic index can be obtained than with homogenous field radiotherapy, but the underlying cellular mechanisms need further understanding. The aim of this study was to investigate the dynamics of radiation-induced bystander effects (RIBE) in rats after exposing one brain hemisphere to either MRT or homogenous synchrotron radiation (HSR). Healthy and tumor-bearing Wistar rats were exposed to doses of 17.5, 35, 70 or 350 Gy, applied either as MRT or HSR to the right cerebral hemisphere. Rats were euthanized at 4, 8 and 12 hours after irradiation to assess the release of bystander signals. Brains and urinary bladders were dissected, and explants for bystander clonogenic reporter assays were set up. Clonogenic survival showed that RIBE occurred in both the non-irradiated brain hemisphere and in bladder of normal and tumor-bearing rats, while the irradiated hemisphere showed the direct effects of radiation. The RIBE observed in our reporter cells shows that both MRT and HSR yield a demonstrable abscopal effect after high doses of irradiation; presumably as part of a systemic response.


Recoverin regulates light-dependent phosphodiesterase activity in retinal rods.
The Ca2+-binding protein recoverin may regulate visual transduction in retinal rods and cones, but its functional role and mechanism of action remain controversial. We compared the photoresponses of rods from control mice and from mice in which the recoverin gene was knocked out. Our analysis indicates that Ca2+-recoverin prolongs the dark-adapted flash response and increases the rod's sensitivity to dim steady light. Knockout rods had faster Ca2+ dynamics, indicating that recoverin is a significant Ca2+ buffer in the outer segment, but incorporation of exogenous buffer did not restore wild-type behavior. We infer that Ca2+-recoverin potentiates light-triggered phosphodiesterase activity, probably by effectively prolonging the catalytic activity of photoexcited rhodopsin.


Reduced ventricular flow propagation velocity in elite athletes is augmented with the resumption of exercise training.
Chronic exercise induces physiological enlargement of the left ventricle ('athlete's heart'), but the effects of current and long-term exercise training on diastolic function have not been investigated. Echocardiography and Doppler imaging were used to assess left ventricular (LV) dimensions and indices of diastolic filling in 22 elite athletes at the end of their 'off-season' (baseline) and, subsequently, following 3 and 6 months of training. Twelve matched controls were also studied at baseline, 3 and 6 months. Compared to controls at baseline, athletes exhibited significantly higher LV mass (235.7 +/- 7.1 g versus 178.1 +/- 14.5 g, P < 0.01) and reduced flow propagation velocity (V(P): 50.21 +/- 1.7 versus 72.2 +/- 3.6 cm s(-1), P < 0.01), a measure of diastolic function. Three months of training further increased LV mass in athletes (253.2 +/- 7.1 g; P < 0.01 versus baseline), and significantly increased their V(P) (66.7 +/- 2.5 cm s(-1); P < 0.05 versus baseline). These trends for increased mass and diastolic filling persisted following 6 months of training (LV mass 249.0 +/- 8.7 g P < 0.05 versus baseline; V(P) 75.7 +/- 3.0 cm s(-1); P < 0.01 versus baseline, and P = 0.01 versus 3 months). This study suggests that following a period of relative inactivity the rate of ventricular relaxation during early diastole may be slowed in athletes who exhibit ventricular hypertrophy, whilst resumption of training increases the speed of ventricular relaxation in the presence of further hypertrophy of the left ventricle.


Isolated repair of the medial patellofemoral ligament in primary dislocation of the patella: a prospective randomized study.
The purpose of this study was to evaluate the effect of surgical reinsertion of the medial patellofemoral ligament (MPFL) to the adductor tubercle compared with conservative treatment in patients with primary dislocation of the patella. Eighty patients with primary patella dislocation were included in the study. Delayed arthroscopy (mean, 50 days after injury) was performed to assess cartilage injury and status. During arthroscopy, patients were randomized to surgical reinsertion of the MPFL or to conservative treatment. Conservative treatment was use of a brace with 0 degrees to 20 degrees motion for the first 2 weeks after dislocation. Patients were followed up for 2 years. The incidence of recurrent patella dislocation was recorded along with the clinical Kujala scores and Knee Injury and Osteoarthritis Outcome Scores. The redislocation rates were 17% and 20% in the operative and conservative treatment groups, respectively (P = not significant). Kujala scores were 85 and 78 in the operative and conservative treatment groups, respectively (P = .07). The patella stability subscore was significantly higher in the operative group. No difference in Knee Injury and Osteoarthritis Outcome Scores was found. Delayed primary repair of the MPFL by use of an anchor-based reattachment to the adductor tubercle without vastus medialis obliquus repair after primary patella dislocation does not reduce the risk of redislocation nor does it produce any significantly better subjective functional outcome based on the Kujala knee score. Only the specific subjective patella stability score was improved by MPFL repair compared with conservative treatment. Level I, therapeutic randomized controlled trial.


The role of lysine-256 in the structure and function of sheep liver recombinant serine hydroxymethyltransferase.
The active site lysine residue, K256, involved in Schiff's base linkage with pyridoxal-5'-phosphate (PLP) in sheep liver recombinant serine hydroxymethyltransferase (rSHMT) was changed to glutamine or arginine by site-directed mutagenesis. The purified K256Q and K256R SHMTs had less than 0.1% of catalytic activity with serine and H4folate as substrates compared to rSHMT. The mutant enzymes also failed to exhibit the characteristic visible absorbance spectrum (lambda(max) 425 nm) and did not produce the quinonoid intermediate (lambda(max) 495 nm) upon the addition of glycine and H4folate. The mutant enzymes were unable to catalyze aldol cleavage of beta-phenylserine and transamination of D-alanine. These results suggested that the mutation of the lysine had resulted in the inability of the enzyme to bind to the cofactor. Therefore, the K256Q SHMT was isolated as a dimer and the K256R SHMT as a mixture of dimers and tetramers which were converted to dimers slowly. On the other hand, rSHMT was stable as a tetramer for several months, further confirming the role of PLP in maintenance of oligomeric structure. The mutant enzymes also failed to exhibit the increased thermal stability upon the addition of serine, normally observed with rSHMT. The enhanced thermal stability has been attributed to a change in conformation of the enzyme from open to closed form leading to reaction specificity. The mutant enzymes were unable to undergo this conformational change probably because of the absence of bound cofactor.


Measurement of viral load by the automated Abbott real-time HIV-1 assay using dried blood spots collected and processed in Malawi and Mozambique.
The use of dried blood spots (DBS) for HIV-1 viral load quantification can greatly improve access to viral monitoring for HIV-infected patients receiving treatment in resource-limited settings. To evaluate and validate HIV viral load measurement from DBS in sub-Saharan Africa, with a reliable, all-automated, standard commercial assay such as the Abbott m2000. A total of 277 DBS were collected in different health centres in Malawi and Mozambique and analysed for viral load determination using the Abbott m2000 assay with the corresponding plasma samples as gold standard. Samples were extracted using the m2000SP automatic extractor and then processed as the plasma samples using the specific 1.0 mL HIV-RNA DBS protocol. Among samples with detectable HIV-RNA the correlation between viral load obtained from the paired 131 plasma and DBS samples was high (r=0.946). Overall, viral load values between DBS and plasma differed by less than 0.5 log unit in 90.1% of cases and by less than 1 log unit in 100% of cases. Using a threshold of 1 000 copies/mL (defining virological failure in resource-limited settings), sensitivity was 94.2% and specificity 98.6%, and both positive and negative predictive values were high (98.5% and 94.5%, respectively). DBS extracted and processed using the Abbott automated system can be reliably used in resource-limited setting to diagnose virological failure.


Global Organization and Proposed Megataxonomy of the Virus World.
Viruses and mobile genetic elements are molecular parasites or symbionts that coevolve with nearly all forms of cellular life. The route of virus replication and protein expression is determined by the viral genome type. Comparison of these routes led to the classification of viruses into seven "Baltimore classes" (BCs) that define the major features of virus reproduction. However, recent phylogenomic studies identified multiple evolutionary connections among viruses within each of the BCs as well as between different classes. Due to the modular organization of virus genomes, these relationships defy simple representation as lines of descent but rather form complex networks. Phylogenetic analyses of virus hallmark genes combined with analyses of gene-sharing networks show that replication modules of five BCs (three classes of RNA viruses and two classes of reverse-transcribing viruses) evolved from a common ancestor that encoded an RNA-directed RNA polymerase or a reverse transcriptase. Bona fide viruses evolved from this ancestor on multiple, independent occasions via the recruitment of distinct cellular proteins as capsid subunits and other structural components of virions. The single-stranded DNA (ssDNA) viruses are a polyphyletic class, with different groups evolving by recombination between rolling-circle-replicating plasmids, which contributed the replication protein, and positive-sense RNA viruses, which contributed the capsid protein. The double-stranded DNA (dsDNA) viruses are distributed among several large monophyletic groups and arose via the combination of distinct structural modules with equally diverse replication modules. Phylogenomic analyses reveal the finer structure of evolutionary connections among RNA viruses and reverse-transcribing viruses, ssDNA viruses, and large subsets of dsDNA viruses. Taken together, these analyses allow us to outline the global organization of the virus world. Here, we describe the key aspects of this organization and propose a comprehensive hierarchical taxonomy of viruses.


Screening for hypothalamic-pituitary-adrenal axis suppression in asthmatic children remains problematic: a cross-sectional study.
To determine which parameter is the most useful screening test for hypothalamic-pituitary-adrenal suppression in asthmatic children. Cross-sectional study. Paediatric allergy clinics in Cape Town, South Africa. 143 asthmatic children of mostly mixed ancestry, aged 5-12 years. Primary outcome measures included Spearman correlation coefficients (r) calculated between the postmetyrapone (PMTP) serum adrenocorticotropic hormone (ACTH), 11-deoxycortisol (11DOC), 11DOC+ cortisol (C) and height, weight, height velocity, weight velocity, change in systolic blood pressure from supine to standing, early morning urinary free cortisol (UFC), morning C, ACTH and dehydroepiandrosterone sulfate (DHEAS). Secondary outcome measures were the receiver operating characteristics (ROC) curve and the diagnostic statistics for the most promising test. All screening variables were weakly correlated with the three PMTP outcomes. Only DHEAS and UFC (nmol/m(2)) were statistically significant-DHEAS for PMTP ACTH and 11DOC (r=0.20, p=0.025 and r=0.21, p=0.017); UFC (nmol/m(2)) for PMTP 11DOC and 11DOC+C (r=0.19, p=0.033 and r=0.20, p=0.022). The area under ROC curve for DHEAS in the 5-year to 9-year age group was 0.69 (95% CI 0.47 to 0.92). At DHEAS cut-off of 0.2 µmol/L: sensitivity=0.88 (CI 0.47 to 1.00), specificity=0.61 (CI 0.42 to 0.78), positive predictive value=0.37 (CI 0.16 to 0.62), negative predictive value=0.95 (CI 0.75 to 1.00), accuracy=0.67 (CI 0.50 to 0.81), positive likelihood ratio=2.26 (CI 1.35 to 3.78), negative likelihood ratio=0.20 (CI 0.03 to 1.30). No parameter is useful as a universal screening test. DHEAS may be suitable to exclude HPAS before adrenarche. Further research is needed to confirm these findings and identify factors, for example, genetic that may predict or protect against HPAS.


Methodology for the independent calibration of Raman backscatter water-vapor lidar systems.
We present a method for the independent calibration of Raman backscatter water-vapor lidar systems. Particular attention is given to the resolution of instrumental changes in the short and the long terms. The method reposes on the decomposition of the instrument function, which allows the lidar calibration coefficient to be re-expressed as the product of two terms, one describing the instrumental transmission and detection efficiency and the other describing the wavelength-dependent convolution of the Raman backscatter cross sections with the instrument function. The origins of changes in instrument response necessitate the experimental determination of the system detection efficiency. Two external light sources for calibration are assessed: zenith observation of diffuse sunlight and a xenon arc lamp. The results favor use of the diffuse-sunlight measurement but highlight the need for simultaneous sunphotometer measurements to constrain modeled aerosol optical properties. Quantum mechanical models of the Raman cross sections are described, and errors in determining the cross sections and their convolution with the instrument function are discussed in detail. The calibration coefficients deduced by using the independent method are compared with coefficients deduced from Vaisala H-Humicap radiosonde measurements. These results agree to within current calibration errors (15%, unconstrained aerosol parameters), and a change in calibration coefficient following instrument modification is reproduced satisfactorily. Results from modeling and intercomparison studies are extended to estimate the calibration accuracy and the precision of the diffuse-sunlight method with constrained modeled aerosol parameters. Changes in the calibration coefficient in the short and the long terms should be resolved to 4(6)% and 6(9)%, respectively, which is comparable or better than the precision of existing dependent methods of calibration. The reduction of the absolute calibration error remains an outstanding issue for all calibration methods.


Amyloid-β peptide-specific DARPins as a novel class of potential therapeutics for Alzheimer disease.
Passive immunization with anti-amyloid-β peptide (Aβ) antibodies is effective in animal models of Alzheimer disease. With the advent of efficient in vitro selection technologies, the novel class of designed ankyrin repeat proteins (DARPins) presents an attractive alternative to the immunoglobulin scaffold. DARPins are small and highly stable proteins with a compact modular architecture ideal for high affinity protein-protein interactions. In this report, we describe the selection, binding profile, and epitope analysis of Aβ-specific DARPins. We further showed their ability to delay Aβ aggregation and prevent Aβ-mediated neurotoxicity in vitro. To demonstrate their therapeutic potential in vivo, mono- and trivalent Aβ-specific DARPins (D23 and 3×D23) were infused intracerebroventricularly into the brains of 11-month-old Tg2576 mice over 4 weeks. Both D23 and 3×D23 treatments were shown to result in improved cognitive performance and reduced soluble Aβ levels. These findings demonstrate the therapeutic potential of Aβ-specific DARPins for the treatment of Alzheimer disease.


Towards a functional neuroanatomy of pleasure and happiness.
The pursuit of happiness is a preoccupation for many people. Yet only the pursuit can be promised, not happiness itself. Can science help? We focus on the most tractable ingredient, hedonia or positive affect. A step toward happiness might be gained by improving the pleasures and positive moods in daily life. The neuroscience of pleasure and reward provides relevant insights, and we discuss how specific hedonic mechanisms might relate to happiness or the lack thereof. Although the neuroscience of happiness is still in its infancy, further advances might be made through mapping overlap between brain networks of hedonic pleasure with others, such as the brain's default network, potentially involved in the other happiness ingredient, eudaimonia or life meaning and engagement.


Asthma exacerbation after administration of nicotine nasal spray for smoking cessation.
A 58-year-old man experienced an asthma exacerbation after administration of nicotine nasal spray for smoking cessation. His medical history was significant for asthma, chronic obstructive pulmonary disease, hypertension, and tobacco use when he was prescribed nicotine nasal spray for smoking cessation in an outpatient setting. Within the first 3 days of administering the spray, he developed wheezing, coughing, and significant shortness of breath, which required hospitalization. The patient was hospitalized for 48 hours and discharged with a diagnosis of asthma exacerbation probably related to administration of nicotine nasal spray. Prudent administration of nicotine nasal spray is recommended in patients with underlying respiratory disease. Patients should be counseled on the potential adverse effects of treatment and proper administration techniques in order to minimize these effects.


Phyllanthus spp. Exerts Anti-Angiogenic and Anti-Metastatic Effects Through Inhibition on Matrix Metalloproteinase Enzymes.
Tumor angiogenesis and metastasis are the major causes for high morbidity and mortality rates in cancer patient. Modulation on tumor angiogenesis and metastasis provides opportunities to halt progression of cancer. From our previous findings, Phyllanthus plant possesses antiproliferative effects on melanoma and prostate cancer cell lines and induction of apoptosis. The main aims of the present work were further investigated on the antimetastatic and antiangiogenic effects on cancer cells (MeWo and PC-3) and human umbilical vein endothelial cells (HUVECs) of 4 Phyllanthus species (P.amarus, P.niruri, P.urinaria and P.watsonii). Phyllanthus extracts significantly inhibited cell adhesion, migration, invasion, and transendothelial migration activities of cancer (MeWo and PC-3) cells in a dose-dependent manner (P < 0.05) by cell-matrix adhesion, Transwell migration, invasion, and transendothelial migration assays. Phyllanthus extracts were exhibited low cytotoxicity on HUVECs up to a concentration of 500.0 μg/ml by MTS reduction assay. Phyllanthus extracts also exhibited antiangiogenic effects through inhibition of migration, invasion, and microcapillary like-tube structure formation in HUVECs. These observations were due to alteration in activities of matrix metalloproteinase (MMP) -2, -7, -9, and -26 in treated-endothelial and cancer cells by zymographies. These findings suggest that Phyllanthus plant has the potential to inhibit tumour metastasis and angiogenesis through the suppression of MMP enzymes.


NU-IN: Nucleotide evolution and input module for the EvolSimulator genome simulation platform.
There is increasing demand to test hypotheses that contrast the evolution of genes and gene families among genomes, using simulations that work across these levels of organization. The EvolSimulator program was developed recently to provide a highly flexible platform for forward simulations of amino acid evolution in multiple related lineages of haploid genomes, permitting copy number variation and lateral gene transfer. Synonymous nucleotide evolution is not currently supported, however, and would be highly advantageous for comparisons to full genome, transcriptome, and single nucleotide polymorphism (SNP) datasets. In addition, EvolSimulator creates new genomes for each simulation, and does not allow the input of user-specified sequences and gene family information, limiting the incorporation of further biological realism and/or user manipulations of the data. We present modified C++ source code for the EvolSimulator platform, which we provide as the extension module NU-IN. With NU-IN, synonymous and non-synonymous nucleotide evolution is fully implemented, and the user has the ability to use real or previously-simulated sequence data to initiate a simulation of one or more lineages. Gene family membership can be optionally specified, as well as gene retention probabilities that model biased gene retention. We provide PERL scripts to assist the user in deriving this information from previous simulations. We demonstrate the features of NU-IN by simulating genome duplication (polyploidy) in the presence of ongoing copy number variation in an evolving lineage. This example is initiated with real genomic data, and produces output that we analyse directly with existing bioinformatic pipelines. The NU-IN extension module is a publicly available open source software (GNU GPLv3 license) extension to EvolSimulator. With the NU-IN module, users are now able to simulate both drift and selection at the nucleotide, amino acid, copy number, and gene family levels across sets of related genomes, for user-specified starting sequences and associated parameters. These features can be used to generate simulated genomic datasets under an extremely broad array of conditions, and with a high degree of biological realism.


Unroofed coronary sinus and coronary sinus orifice atresia. Implications for management of complex congenital heart disease.
The aim of this study was to assess the morphology of the coronary sinus, its drainage and associated cardiac malformations when there is either complete unroofing of the coronary sinus or atresia of its connection to the right atrium. As more children with complex cardiac anomalies are accepted for primary surgical repair or palliation with cavopulmonary anastomoses, a knowledge of coronary sinus and systemic venous anomalies is important if coronary venous return is to be preserved and residual shunts avoided. Twenty-six heart-lung specimens without a coronary sinus draining to the right atrium were identified from the Leiden collection of congenital heart malformations. These were classified into specimens with an unroofed coronary sinus and those with atresia of the coronary sinus orifice. Attention was paid to the associated cardiac malformations. In 14 (54%; confidence limits [CL] 35%, 73%) of 26 specimens, there was an unroofed coronary sinus, associated with persistence of the left superior caval vein. An inferoposterior location of an atrial septal defect was detected in 2 (14%; CL -4%, 33%) of 14. Atrial appendage anomalies were seen in 13 (93%; CL 79%, 106%) of 14 specimens, exemplified by both right and left isomerism. These were frequently associated with an atrioventricular septal defect (12 [86%; CL 67%, 104%] of 14). An atretic coronary sinus orifice was seen in 12 (46%; CL 27%, 65%) of 26. Atrial appendage anomalies (2 [17%; CL -4%, 38%] of 12) were rare in these cases. The drainage was then by way of a left superior caval vein or, in its absence, a coronary sinus to left atrial window. Ventricular hypoplasia was seen in both categories of coronary sinus abnormalities. Important ventricular hypoplasia was seen in 12 cases (46%; CL 27%, 65%). These findings emphasize the need to study coronary sinus drainage before procedures such as ligation or transcatheter coil embolization of a left superior caval vein, venous redirection or closure of a dorsal atrial septal defect are contemplated. These procedures might inadvertently lead to impairment of coronary venous return or persistence of an intracardiac shunt.


Involvement of histone H1 in the organization of the nucleosome and of the salt-dependent superstructures of chromatin.
We describe the results of a systematic study, using electron microscopy, of the effects of ionic strength on the morphology of chromatin and of H1-depleted chromatin. With increasing ionic strength, chromatin folds up progressively from a filament of nucleosomes at approximately 1 mM monovalent salt through some intermediate higher-order helical structures (Thoma, F., and T. Koller, 1977, Cell 12:101-107) with a fairly constant pitch but increasing numbers of nucleosomes per turn, until finally at 60 mM (or else in approximately 0.3 mM Mg++) a thick fiber of 250 A diameter is formed, corresponding to a structurally well-organized but not perfectly regular superhelix or solenoid of pitch approximately 110 A as described by Finch and Klug (1976, Proc. Natl. Acad. Sci. U.S.A. 73:1897-1901). The numbers of nucleosomes per turn of the helical structures agree well with those which can be calculated from the light-scattering data of Campbell et al. (1978, Nucleic Acids Res. 5:1571-1580). H1-depleted chromatin also condenses with increasing ionic strength but not so densely as chromatin and not into a definite structure with a well-defined fiber direction. At very low ionic strengths, nucleosomes are present in chromatin but not in H1-depleted chromatin which has the form of an unravelled filament. At somewhat higher ionic strengths (greater than 5 mM triethanolamine chloride), nucleosomes are visible in both types of specimen but the fine details are different. In chromatin containing H1, the DNA enters and leaves the nucleosome on the same side but in chromatin depleted of H1 the entrance and exit points are much more random and more or less on opposite sides of the nucleosome. We conclude that H1 stabilizes the nucleosome and is located in the region of the exit and entry points of the DNA. This result is correlated with biochemical and x-ray crystallographic results on the internal structure of the nucleosome core to give a picture of a nucleosome in which H1 is bound to the unique region on a complete two-turn, 166 base pair particle (Fig. 15). In the formation of higher-order structures, these regions on neighboring nucleosomes come closer together so that an H1 polymer may be formed in the center of the superhelical structures.


A prospective investigation of interleukin-8 levels in pediatric acute respiratory failure and acute respiratory distress syndrome.
The association of plasma interleukin-8 (IL-8), or IL-8 genetic variants, with pediatric acute respiratory distress syndrome (PARDS) in children with acute respiratory failure at risk for PARDS has not been examined. The purpose of this study was to examine the association of early and sequential measurement of plasma IL-8 and/or its genetic variants with development of PARDS and other clinical outcomes in mechanically ventilated children with acute respiratory failure. This was a prospective cohort study of children 2 weeks to 17 years of age with acute airways and/or parenchymal lung disease done in 22 pediatric intensive care units participating in the multi-center clinical trial, Randomized Evaluation of Sedation Titration for Respiratory Failure (RESTORE). Plasma IL-8 levels were measured within 24 h of consent and 24 and 48 h later. DNA was purified from whole blood, and IL-8 single nucleotide polymorphisms, rs4073, rs2227306, and rs2227307, were genotyped. Five hundred forty-nine patients were enrolled; 480 had blood sampling. Plasma IL-8 levels ranged widely from 4 to 7373 pg/mL. Highest IL-8 levels were observed on the day of intubation with subsequent tapering. Levels of IL-8 varied significantly across primary diagnoses with the highest levels occurring in patients with sepsis and the lowest levels in those with asthma. Plasma IL-8 was strongly correlated with oxygenation defect and severity of illness. IL-8 was consistently higher in PARDS patients compared to those without PARDS; levels were 4-12 fold higher in non-survivors compared to survivors. On multivariable analysis, IL-8 was independently associated with death, duration of mechanical ventilation, and PICU length of stay on all days measured, but was not associated with PARDS development. There was no association between the IL-8 single nucleotide polymorphisms, rs4073, rs2227306, and rs2227307, and PARDS development or plasma IL-8 level. When measured sequentially, plasma IL-8 was robustly associated with multiple, relevant clinical outcomes including mortality, but was not associated with PARDS development. The wide range of plasma IL-8 levels exhibited in this cohort suggests that further study into the heterogeneity of this patient population and its impact on individual responses to PARDS treatment is warranted.


Astrocytes contribute to remote memory formation by modulating hippocampal-cortical communication during learning.
Remote memories depend on coordinated activity in the hippocampus and frontal cortices, but the timeline of these interactions is debated. Astrocytes sense and modify neuronal activity, but their role in remote memory is scarcely explored. We expressed the Gi-coupled designer receptor hM4Di in CA1 astrocytes and discovered that astrocytic manipulation during learning specifically impaired remote, but not recent, memory recall and decreased activity in the anterior cingulate cortex (ACC) during retrieval. We revealed massive recruitment of ACC-projecting CA1 neurons during memory acquisition, which was accompanied by the activation of ACC neurons. Astrocytic Gi activation disrupted CA3 to CA1 communication in vivo and reduced the downstream response in the ACC. In behaving mice, it induced a projection-specific inhibition of CA1-to-ACC neurons during learning, which consequently prevented ACC recruitment. Finally, direct inhibition of CA1-to-ACC-projecting neurons spared recent and impaired remote memory. Our findings suggest that remote memory acquisition involves projection-specific functions of astrocytes in regulating CA1-to-ACC neuronal communication.


Structure of a signal transduction regulator, RACK1, from Arabidopsis thaliana.
The receptor for activated C-kinase 1 (RACK1) is a highly conserved WD40 repeat scaffold protein found in a wide range of eukaryotic species from Chlamydymonas to plants and humans. In tissues of higher mammals, RACK1 is ubiquitously expressed and has been implicated in diverse signaling pathways involving neuropathology, cellular stress, protein translation, and developmental processes. RACK1 has established itself as a scaffold protein through physical interaction with a myriad of signaling proteins ranging from kinases, phosphatases, ion channels, membrane receptors, G proteins, IP3 receptor, and with widely conserved structural proteins associated with the ribosome. In the plant Arabidopsis thaliana, RACK1A is implicated in diverse developmental and environmental stress pathways. Despite the functional conservation of RACK1-mediated protein-protein interaction-regulated signaling modes, the structural basis of such interactions is largely unknown. Here we present the first crystal structure of a RACK1 protein, RACK1 isoform A from Arabidopsis thaliana, at 2.4 A resolution, as a C-terminal fusion of the maltose binding protein. The structure implicates highly conserved surface residues that could play critical roles in protein-protein interactions and reveals the surface location of proposed post-transcriptionally modified residues. The availability of this structure provides a structural basis for dissecting RACK1-mediated cellular signaling mechanisms in both plants and animals.


Evaluation of autologous platelet-poor plasma gel as a hemostatic adjunct after posterior iliac crest bone harvest.
To evaluate the hemostatic efficacy of autologous platelet-poor plasma (PPP) gel following posterior iliac crest bone graft harvesting for oral and maxillofacial reconstruction. This was a prospective study of 24 consecutive patients involving 26 posterior iliac crest bone harvests that had bone wax and either 1-gram of bovine microfibrillar collagen or 20 mL of autologous PPP, activated as a gel, used as adjunct hemostatic agents. Compression bulb suction drain was placed into the graft site and drain output recorded every 8 hours for 64 hours. Cost analysis was also undertaken between the 2 methods. Statistical significance between means of each group was determined by Student's t test and found significant for P < .05. There were no statistically significant differences in average drain output between the PPP and MFC groups for each 8-hour interval. There was no statically significant difference in average total drain output between the PPP and MFC groups over the entire 64 hour period. Additionally, unlike the addition of MFC, the addition of PPP added no additional costs to the procedure. PPP gel, when compared with bovine microfibrillar collagen, is an effective and inexpensive adjunct in hemostasis following posterior iliac crest bone harvest.


Anodal cerebellar t-DCS impacts skill learning and transfer on a robotic surgery training task.
The cerebellum has demonstrated a critical role during adaptation in motor learning. However, the extent to which it can contribute to the skill acquisition of complex real-world tasks remains unclear. One particularly challenging application in terms of motor activities is robotic surgery, which requires surgeons to complete complex multidimensional visuomotor tasks through a remotely operated robot. Given the need for high skill proficiency and the lack of haptic feedback, there is a pressing need for understanding and improving skill development. We investigated the effect of cerebellar transcranial direct current stimulation applied during the execution of a robotic surgery training task. Study participants received either real or sham stimulation while performing a needle driving task in a virtual (simulated) and a real-world (actual surgical robot) setting. We found that cerebellar stimulation significantly improved performance compared to sham stimulation at fast (more demanding) execution speeds in both virtual and real-world training settings. Furthermore, participants that received cerebellar stimulation more effectively transferred the skills they acquired during virtual training to the real world. Our findings underline the potential of non-invasive brain stimulation to enhance skill learning and transfer in real-world relevant tasks and, more broadly, its potential for improving complex motor learning.


Utility of both muscle and fascia flaps in severe lower extremity trauma.
The evolving technology in trauma management today permits salvage of many severe lower extremity injuries previously even considered to be lethal. An essential component for any such treatment protocol must be adequate soft tissue coverage that often will use vascularized flaps. Traditionally, calf muscles have been used proximally and free flaps for the distal leg and foot. The reintroduction of reliable local fascia flaps has challenged this dictum, proving to be a simpler and yet versatile option. The role of both muscle and fascia flaps in lower extremity injuries has been retrospectively reviewed from a 2-decade experience. Soft tissue deficits requiring some form of vascularized flap occurred in 160 limbs in 155 patients. The frequency of use of flap types, specific complications and benefits, effect of timing of wound closure, and rate of limb salvage were compared. Initial coverage after significant lower extremity trauma in these 160 limbs required 60 local muscle flaps, 50 local fascia flaps, and 74 free flaps. These flaps had been selected on a nonrandom basis according to wound location, its severity, and flap availability. Complications were directly related to the severity of injury, and for free flaps as a group (39%), although these were not independent variables. Local muscle (27%) or fascia flaps (30%) were similar with regard to this morbidity. Healing was more likely to be uneventful if coverage were accomplished during the acute period after injury, regardless of flap type. Muscle flaps were still used in two thirds of all cases, with the soleus muscle used as often for the distal leg as the mid-leg. Local fascia flaps were most valuable for smaller defects, especially in the distal leg or foot, and often as a reasonable alternative to a free flap. The traditional role of the gastrocnemius muscles for flap coverage of knee and proximal leg defects and the soleus muscle for the middle third of the leg was reaffirmed. The soleus muscle often also reached distal leg defects as could local fascia flaps, where classically, otherwise, a free flap would have been necessary. The largest or most severe wounds, irrespective of limb location, required free flap coverage. Local fascia flaps proved to be a valuable alternative.


Gene transcription in Daphnia magna: effects of acute exposure to a carbamate insecticide and an acetanilide herbicide.
Daphnia magna is a key invertebrate in the freshwater environment and is used widely as a model in ecotoxicological measurements and risk assessment. Understanding the genomic responses of D. magna to chemical challenges will be of value to regulatory authorities worldwide. Here we exposed D. magna to the insecticide methomyl and the herbicide propanil to compare phenotypic effects with changes in mRNA expression levels. Both pesticides are found in drainage ditches and surface water bodies standing adjacent to crops. Methomyl, a carbamate insecticide widely used in agriculture, inhibits acetylcholinesterase, a key enzyme in nerve transmission. Propanil, an acetanilide herbicide, is used to control grass and broad-leaf weeds. The phenotypic effects of single doses of each chemical were evaluated using a standard immobilisation assay. Immobilisation was linked to global mRNA expression levels using the previously estimated 48h-EC(1)s, followed by hybridization to a cDNA microarray with more than 13,000 redundant cDNA clones representing >5000 unique genes. Following exposure to methomyl and propanil, differential expression was found for 624 and 551 cDNAs, respectively (one-way ANOVA with Bonferroni correction, P</=0.05, more than 2-fold change) and up-regulation was prevalent for both test chemicals. Both pesticides promoted transcriptional changes in energy metabolism (e.g., mitochondrial proteins, ATP synthesis-related proteins), moulting (e.g., chitin-binding proteins, cuticular proteins) and protein biosynthesis (e.g., ribosomal proteins, transcription factors). Methomyl induced the transcription of genes involved in specific processes such as ion homeostasis and xenobiotic metabolism. Propanil highly promoted haemoglobin synthesis and up-regulated genes specifically related to defence mechanisms (e.g., innate immunity response systems) and neuronal pathways. Pesticide-specific toxic responses were found but there is little evidence for transcriptional responses purely restricted to genes associated with the pesticide target site or mechanism of toxicity.


In vivo transplantation of fetal human gut-derived enteric neural crest cells.
The prospect of using neural cell replacement for the treatment of severe enteric neuropathies has seen significant progress in the last decade. The ability to harvest and transplant enteric neural crest cells (ENCCs) that functionally integrate within recipient intestine has recently been confirmed by in vivo murine studies. Although similar cells can be harvested from human fetal and postnatal gut, no studies have as yet verified their functional viability upon in vivo transplantation. We sought to determine whether ENCCs harvested from human fetal bowel are capable of engraftment and functional integration within recipient intestine following in vivo transplantation into postnatal murine colon. Enteric neural crest cells selected and harvested from fetal human gut using the neurotrophin receptor p75NTR were lentivirally labeled with either GFP or calcium-sensitive GCaMP and transplanted into the hindgut of Rag2- /γc- /C5- -immunodeficient mice at postnatal day 21. Transplanted intestines were assessed immunohistochemically for engraftment and differentiation of donor cells. Functional viability and integration with host neuromusculature was assessed using calcium imaging. Transplanted human fetal gut-derived ENCC showed engraftment within the recipient postnatal colon in 8/15 mice (53.3%). At 4 weeks posttransplantation, donor cells had spread from the site of transplantation and extended projections over distances of 1.2 ± 0.6 mm (n = 5), and differentiated into enteric nervous system (ENS) appropriate neurons and glia. These cells formed branching networks located with the myenteric plexus. Calcium transients (change in intensity F/F0 = 1.25 ± 0.03; 15 cells) were recorded in transplanted cells upon stimulation of the recipient endogenous ENS demonstrating their viability and establishment of functional connections.


Pharmacokinetic studies of 5-fluorouracil and 5'-deoxy-5-fluorouridine in rats.
The pharmacokinetics of 5-fluorouracil (FU) and its metabolic prodrug, 5'-deoxy-5-fluorouridine (dFUR), were investigated in 5- to 9-month-old rats without tumors. Intravenous bolus injections and infusions of FU (25-35 mg X kg-1) and dFUR (500-750 mg X kg-1) had activity against transplanted colon tumors in 2- to 4-month-old rats. Blood and plasma concentrations of FU and dFUR were analyzed by high-pressure liquid chromatography using 5-bromouracil as the internal standard. Free fractions of these drugs in plasma were indistinguishable from unity, indicating little or no protein binding. The area under the blood concentration-time profiles and the steady-state concentrations of unchanged dFUR were 40- to 100-fold higher than those of unchanged FU. The respective blood clearances of FU and dFUR were 44 and 18 ml X kg-1 X min-1 after i.v. bolus injections, which were significantly lower than the 108 and 26 ml X kg-1 X min-1 after infusion. FU clearance decreased when its infusion rate was increased from 25 to 50 mg X kg-1 X day-1. Renal clearances of both drugs remained the same after either route of administration. These data suggest that both drugs were eliminated by nonlinear kinetics and that their metabolism was saturated at a more rapid administration rate.(ABSTRACT TRUNCATED AT 250 WORDS)


[Hyperbaric oxygenation in neurotraumatology].
The use of hyperbaric oxygenation (HBO) as a therapeutic measure has a favorable effect, in the postoperative period also, on traumatic affections of different levels of the nervous system (the brain, spinal cord, peripheral nerve trunks and plexuses. It allows the severity of the neurologic deficit and the degree of the patient's disability to be reduced and in some cases it prevents the growth of vital disorders. At the same time, the effect may exert a certain load on the adaptation mechanisms which are very limited in many neurosurgical patients. In view of this, the tolerance of the patient's organism to this load must be taken into account in choosing the HBO schedules. The tolerance is decisively determined by the intactness of the integral function of the central nervous system and diminishes markedly as the impairment of the consciousness of patients with primary affection of the brain gradually becomes deeper and longer in duration.


Analytic signal phase-based myocardial motion estimation in tagged MRI sequences by a bilinear model and motion compensation.
Different mathematical tools, such as multidimensional analytic signals, allow for the calculation of 2D spatial phases of real-value images. The motion estimation method proposed in this paper is based on two spatial phases of the 2D analytic signal applied to cardiac sequences. By combining the information of these phases issued from analytic signals of two successive frames, we propose an analytical estimator for 2D local displacements. To improve the accuracy of the motion estimation, a local bilinear deformation model is used within an iterative estimation scheme. The main advantages of our method are: (1) The phase-based method allows the displacement to be estimated with subpixel accuracy and is robust to image intensity variation in time; (2) Preliminary filtering is not required due to the bilinear model. The proposed algorithm, integrating phase-based optical flow motion estimation and the combination of global motion compensation with local bilinear transform, allows spatio-temporal cardiac motion analysis, e.g. strain and dense trajectory estimation over the cardiac cycle. Results from 7 realistic simulated tagged magnetic resonance imaging (MRI) sequences show that our method is more accurate compared with state-of-the-art method for cardiac motion analysis and with another differential approach from the literature. The motion estimation errors (end point error) of the proposed method are reduced by about 33% compared with that of the two methods. In our work, the frame-to-frame displacements are further accumulated in time, to allow for the calculation of myocardial Lagrangian cardiac strains and point trajectories. Indeed, from the estimated trajectories in time on 11 in vivo data sets (9 patients and 2 healthy volunteers), the shape of myocardial point trajectories belonging to pathological regions are clearly reduced in magnitude compared with the ones from normal regions. Myocardial point trajectories, estimated from our phase-based analytic signal approach, seem therefore a good indicator of the local cardiac dynamics. Moreover, they are shown to be coherent with the estimated deformation of the myocardium.


Interleukin-23: a promising therapeutic target in seronegative spondyloarthropathy.
Particular therapeutic challenges are raised by the spondyloarthropathies which represent a key area of unmet medical need. Recent investigations have shown that these conditions are characterised both by altered responsiveness to interleukin(IL)-23 and expansion of IL-23 responsive cells as well as increased production of IL-23. The gut in particular has emerged as a key site of IL-23 production, and gut inflammation is known to be strongly clinically associated with these conditions. Moreover, HLA-B27, which is strongly associated with spondyloarthropathy, has also been shown to stimulate IL-23 production. The view is thus emerging that dysregulation of IL-23 biology is a unifying feature of spondyloarthropathy, suggesting that treatments targeting this cytokine are likely to be highly efficacious.


Fructose and Cardiometabolic Health: What the Evidence From Sugar-Sweetened Beverages Tells Us.
Recent attention has focused on fructose as having a unique role in the pathogenesis of cardiometabolic diseases. However, because we rarely consume fructose in isolation, the major source of fructose in the diet comes from fructose-containing sugars, sucrose and high fructose corn syrup, in sugar-sweetened beverages and foods. Intake of sugar-sweetened beverages has been consistently linked to increased risk of obesity, type 2 diabetes, and cardiovascular disease in various populations. Putative underlying mechanisms include incomplete compensation for liquid calories, adverse glycemic effects, and increased hepatic metabolism of fructose leading to de novo lipogenesis, production of uric acid, and accumulation of visceral and ectopic fat. In this review we summarize the epidemiological and clinical trial evidence evaluating added sugars, especially sugar-sweetened beverages, and the risk of obesity, diabetes, and cardiovascular disease and address potential biological mechanisms with an emphasis on fructose physiology. We also discuss strategies to reduce intake of fructose-containing beverages.


A comprehensive network and pathway analysis of candidate genes in major depressive disorder.
Numerous genetic and genomic datasets related to complex diseases have been made available during the last decade. It is now a great challenge to assess such heterogeneous datasets to prioritize disease genes and perform follow up functional analysis and validation. Among complex disease studies, psychiatric disorders such as major depressive disorder (MDD) are especially in need of robust integrative analysis because these diseases are more complex than others, with weak genetic factors at various levels, including genetic markers, transcription (gene expression), epigenetics (methylation), protein, pathways and networks. In this study, we proposed a comprehensive analysis framework at the systems level and demonstrated it in MDD using a set of candidate genes that have recently been prioritized based on multiple lines of evidence including association, linkage, gene expression (both human and animal studies), regulatory pathway, and literature search. In the network analysis, we explored the topological characteristics of these genes in the context of the human interactome and compared them with two other complex diseases. The network topological features indicated that MDD is similar to schizophrenia compared to cancer. In the functional analysis, we performed the gene set enrichment analysis for both Gene Ontology categories and canonical pathways. Moreover, we proposed a unique pathway crosstalk approach to examine the dynamic interactions among biological pathways. Our pathway enrichment and crosstalk analyses revealed two unique pathway interaction modules that were significantly enriched with MDD genes. These two modules are neuro-transmission and immune system related, supporting the neuropathology hypothesis of MDD. Finally, we constructed a MDD-specific subnetwork, which recruited novel candidate genes with association signals from a major MDD GWAS dataset. This study is the first systematic network and pathway analysis of candidate genes in MDD, providing abundant important information about gene interaction and regulation in a major psychiatric disease. The results suggest potential functional components underlying the molecular mechanisms of MDD and, thus, facilitate generation of novel hypotheses in this disease. The systems biology based strategy in this study can be applied to many other complex diseases.


Influence of halide binding on the hydrogen bonding network in the active site of Salinibacter sensory rhodopsin I.
In nature, organisms are subjected to a variety of environmental stimuli to which they respond and adapt. They can show avoidance or attractive behaviors away from or toward such stimuli in order to survive in the various environments in which they live. One such stimuli is light, to which, for example, the receptor sensory rhodopsin I (SRI) has been found to respond by regulating both negative and positive phototaxis in, e.g., the archaeon Halobacterium salinarum. Interestingly, to date, all organisms having SRI-like proteins live in highly halophilic environments, suggesting that salt significantly influences the properties of SRIs. Taking advantage of the discovery of the highly stable SRI homologue from Salinibacter ruber (SrSRI), which maintains its color even in the absence of salt, the importance of the chloride ion for the color tuning and for the slow M-decay, which is thought to be essential for the phototaxis function of SRIs, has been reported previously [Suzuki, D., et al. (2009) J. Mol. Biol.392, 48-62]. Here the effects of the anion binding on the structure and structural changes of SRI during its photocycle are investigated by means of Fourier transform infrared (FTIR) spectroscopy and electrochemical experiments. Our results reveal that, among other things, the structural change and proton movement of a characteristic amino acid residue, Asp102 in SrSRI, is suppressed by the binding of an anion in its vicinity, both in the K- and M-intermediate. The presence of this anion also effects the extent of chromophore distrotion, and tentative results indicate an influence on the number and/or properties of internal water molecules. In addition, a photoinduced proton transfer could only be observed in the absence of the bound anion. Possible proton movement pathways, including the residues Asp102 and the putative Cl binding site His131, are discussed. In conclusion, the results show that the anion binding to SRI is not only important for the color tuning, and for controlling the photocycle kinetics, but also induces some structural changes which facilitate the observed properties.


Complete sequence of RNA1 and subgenomic RNA3 of Atlantic halibut nodavirus (AHNV).
The Nodaviridae are divided into the alphanodavirus genus, which infects insects, and the betanodavirus genus, which infects fishes. Betanodaviruses are the causative agent of viral encephalopathy and retinopathy (VER) in a number of cultivated marine fish species. The Nodaviridae are small non-enveloped RNA viruses that contain a genome consisting of 2 single-stranded positivesense RNA segments: RNA1 (3.1 kb), which encodes the viral part of the RNA-dependent RNA polymerase (RdRp); and RNA2 (1.4 kb), which encodes the capsid protein. In addition to RNA1 and RNA2, a subgenomic transcript of RNA1, RNA3, is present in infected cells. We have cloned and sequenced RNA1 from the Atlantic halibut Hippoglossus hippoglossus nodavirus (AHNV), and for the first time, the sequence of a betanodaviral subgenomic RNA3 has been determined. AHNV RNA1 was 3100 nucleotides in length and contained a main open reading frame encoding a polypeptide of 981 amino acids. Conservative motifs for RdRp were found in the deduced amino acid sequence. RNA3 was 371 nucleotides in length, and contained an open reading frame encoding a peptide of 75 amino acids corresponding to a hypothetical B2 protein, although sequence alignments with the alphanodavirus B2 proteins showed only marginal similarities. AHNV RNA replication in the fish cell-line SSN-1 (derived from striped snakehead) was analysed by Northern blot analysis, which indicated that RNA3 was synthesised in large amounts (compared to RNA1) at an early point in time post-infection.


Phosphorylated KDR can be located in the nucleus of neoplastic cells.
KDR (kinase insert domain receptor) phosphorylation induces several effects which lead eventually to cell proliferation and survival. The precise mechanisms by which KDR, once it is activated, communicates with the nucleus are starting to be understood but have not yet been completely unravelled. Two in vitro studies on animal cell lines reported in the literature have demonstrated that, following stimulation with VEGF, KDR is actually translocated within the nucleus. Our aim was to investigate whether this translocation occurs in human cells both in vitro and in vivo. Using laser scanning confocal microscopy, a variable nuclear localization of phosphorylated and total KDR in cell lines and tumour samples was found. In human neoplastic cell lines, hypoxic stimulation greatly increased the nuclear amount of total KDR but less so that of the phosphorylated form. Only after hypoxia and VEGF stimulation there was a comparably increased expression of phosphorylated and total KDR observed in the nuclei of these cells. We conclude that neoplastic cells show a variable expression of total and phosphorylated KDR in the nucleus. The precise functional meaning of nuclear location remains to be established.


Dynamic contrast-enhanced magnetic resonance imaging perfusion characteristics in meningiomas treated with resection and adjuvant radiosurgery.
OBJECTIVEThere is a need for advanced imaging biomarkers to improve radiation treatment planning and response assessment. T1-weighted dynamic contrast-enhanced perfusion MRI (DCE MRI) allows quantitative assessment of tissue perfusion and blood-brain barrier dysfunction and has entered clinical practice in the management of primary and secondary brain neoplasms. The authors sought to retrospectively investigate DCE MRI parameters in meningiomas treated with resection and adjuvant radiation therapy using volumetric segmentation.METHODSA retrospective review of more than 300 patients with meningiomas resected between January 2015 and December 2018 identified 14 eligible patients with 18 meningiomas who underwent resection and adjuvant radiotherapy. Patients were excluded if they did not undergo adjuvant radiation therapy or DCE MRI. Demographic and clinical characteristics were obtained and compared to DCE perfusion metrics, including mean plasma volume (vp), extracellular volume (ve), volume transfer constant (Ktrans), rate constant (kep), and wash-in rate of contrast into the tissue, which were derived from volumetric analysis of the enhancing volumes of interest.RESULTSThe mean patient age was 64 years (range 49-86 years), and 50% of patients (7/14) were female. The average tumor volume was 8.07 cm3 (range 0.21-27.89 cm3). The median Ki-67 in the cohort was 15%. When stratified by median Ki-67, patients with Ki-67 greater than 15% had lower median vp (0.02 vs 0.10, p = 0.002), and lower median wash-in rate (1.27 vs 4.08 sec-1, p = 0.04) than patients with Ki-67 of 15% or below. Logistic regression analysis demonstrated a statistically significant, moderate positive correlation between ve and time to progression (r = 0.49, p < 0.05). Furthermore, there was a moderate positive correlation between Ktrans and time to progression, which approached, but did not reach, statistical significance (r = 0.48, p = 0.05).CONCLUSIONSThis study demonstrates a potential role for DCE MRI in the preoperative characterization and stratification of meningiomas, laying the foundation for future prospective studies incorporating DCE as a biomarker in meningioma diagnosis and treatment planning.


Care of Incarcerated Patients.
The United States has the highest incarceration rate of any nation in the world. Approximately 870 of every 100,000 U.S. citizens are currently in jails or prisons. U.S. inmates are disproportionately young males, racial and ethnic minorities, and persons of low socioeconomic status. Incarcerated persons have high rates of psychiatric conditions, communicable diseases, substance use disorders, and chronic diseases. The U.S. Preventive Services Task Force recommends that all inmates be screened for human immunodeficiency virus infection, hepatitis C, syphilis, and latent tuberculosis infection, and that sexually active female inmates be screened for gonorrhea and chlamydia. Inmates should also be screened for psychiatric conditions and substance use disorders. Therapy should be continued for all chronic conditions when indicated. Inmates should be referred to community organizations for follow-up medical care and treatment of substance use disorders before they are released from detention facilities. A systematic approach to urgent, routine, and preventive care for persons in jails and prisons creates a healthier correctional environment and a healthier community after release.


Specifying content and mechanisms of change in interventions to change professionals' practice: an illustration from the Good Goals study in occupational therapy.
It is widely agreed that interventions to change professionals' practice need to be clearly specified. This involves (1) selecting and defining the intervention techniques, (2) operationalising the techniques and deciding their delivery, and (3) formulating hypotheses about the mechanisms through which the techniques are thought to result in change. Descriptions of methods to achieve these objectives are limited. This paper reports methods and illustrates outputs from a study to meet these objectives, specifically from the Good Goals study to improve occupational therapists' caseload management practice. (1) Behaviour change techniques were identified and selected from an existing matrix that maps techniques to determinants. An existing coding manual was used to define the techniques. (2) A team of occupational therapists generated context-relevant, acceptable modes of delivery for the techniques; these data were compared and contrasted with previously collected data, literature on caseload management, and the aims of the intervention. (3) Hypotheses about the mechanisms of change were formulated by drawing on the matrix and on theories of behaviour change. (1) Eight behaviour change techniques were selected: goal specified; self-monitoring; contract; graded tasks; increasing skills (problem solving, decision making, goal setting); coping skills; rehearsal of relevant skills; social processes of encouragement, support, and pressure; demonstration by others; and feedback. (2) A range of modes of delivery were generated (e.g., graded tasks' consisting of series of clinical cases and situations that become increasingly difficult). Conditions for acceptable delivery were identified (e.g., 'self-monitoring' was acceptable only if delivered at team level). The modes of delivery were specified as face-to-face training, task sheets, group tasks, DVDs, and team-based weekly meetings. (3) The eight techniques were hypothesized to target caseload management practice through eleven mediating variables. Three domains were hypothesized to be most likely to change: beliefs about capabilities, motivation and goals, and behavioural regulation. The project provides an exemplar of a systematic and reportable development of a quality-improvement intervention, with its methods likely to be applicable to other projects. A subsequent study of the intervention has provided early indication that use of systematic methods to specify interventions may help to maximize acceptability and effectiveness.


Genetic incorporation of the protein transduction domain of Tat into Ad5 fiber enhances gene transfer efficacy.
Human adenovirus serotype 5 (Ad5) has been widely explored as a gene delivery vector for a variety of diseases. Many target cells, however, express low levels of Ad5 native receptor, the Coxsackie-Adenovirus Receptor (CAR), and thus are resistant to Ad5 infection. The Protein Transduction Domain of the HIV Tat protein, namely PTD tat, has been shown to mediate protein transduction in a wide range of cells. We hypothesize that re-targeting Ad5 vector via the PTD tat motif would improve the efficacy of Ad5-mediated gene delivery. In this study, we genetically incorporated the PTD tat motif into the knob domain of Ad5 fiber, and rescued the resultant viral vector, Ad5.PTD tat. Our data showed the modification did not interfere with Ad5 binding to its native receptor CAR, suggesting Ad5 infection via the CAR pathway is retained. In addition, we found that Ad5.PTD tat exhibited enhanced gene transfer efficacy in all of the cell lines that we have tested, which included both low-CAR and high-CAR decorated cells. Competitive inhibition assays suggested the enhanced infectivity of Ad5.PTD tat was mediated by binding of the positively charged PTD tat peptide to the negatively charged epitopes on the cells' surface. Furthermore, we investigated in vivo gene delivery efficacy of Ad5.PTD tat using subcutaneous tumor models established with U118MG glioma cells, and found that Ad5.PTD tat exhibited enhanced gene transfer efficacy compared to unmodified Ad5 vector as analyzed by a non-invasive fluorescence imaging technique. Genetic incorporation of the PTD tat motif into Ad5 fiber allowed Ad5 vectors to infect cells via an alternative PTD tat targeting motif while retaining the native CAR-mediated infection pathway. The enhanced infectivity was demonstrated in both cultured cells and in in vivo tumor models. Taken together, our study identifies a novel tropism expanded Ad5 vector that may be useful for clinical gene therapy applications.


CdSe nanocrystal sensitized photon upconverting film.
Here, films using CdSe nanocrystal (NC) triplet photosensitizers in conjunction with diphenylanthracene (DPA) emitters were assembled to address several challenges to practical applications for solution-based photon upconversion. By using poly(9-vinylcarbazole) as a phosphorescent host in this film, volatile organic solvents are eliminated, the spontaneous crystallization of the emitter is significantly retarded, and ∼1.5% photon upconversion quantum yield (out of a maximum of 50%) is obtained. Transient absorption spectroscopy on nanosecond-to-microsecond time scales reveals this efficiency is enabled by an exceptionally long triplet lifetime of 3.4 ± 0.3 ms. Ultimately, we find the upconversion efficiency is limited by incomplete triplet-triplet annihilation, which occurs with a rate 3-4 orders of magnitude slower than in solution-phase upconversion systems.


Value and limitations of the electrocardiographic response to exercise in the assessment of patients with coronary artery disease. Controversies in cardiology--II.
The results of exercise testing provide a probabibity statement rather than a definitive answer regarding the existence of coronary disease. Moreover, in asymptomatic subjects (1) the S-T segment response to exercise testing has a relatively poor predictive accuracy, (2) exercise testing does not appear to be a practical screening method for detecting subjects at high risk of sudden death, and (3) risk factors should be vigorously treated regardless of the results of screening studies for coronary artery disease. Hence, the probability statements derived from the electrocardiographic response to exercise testing do not appear to provide important enough diagnostic or prognostic information to justify routine use in asymptomatic subjects. Although exercise screening studies provide only marginal aid in detection coronary artery disease in symptomatic patients, important prognostic information may be derived that could influence decisions regarding pharmacologic or surgical therapy. Thus, although electrocardiographic stress testing should not be used as a routine screening procedure, its judicious use in symptomatic patients may provide the physician with important information for patient management.


Feasibility of Real-Time Near-Infrared Fluorescence Tracer Imaging in Sentinel Node Biopsy for Oral Cavity Cancer Patients.
Sentinel node biopsy (SNB) is an established method in oral squamous cell carcinoma (OSCC) for staging the cN0 neck and to select patients who will benefit from a neck dissection. Near-infrared fluorescence (NIRF) imaging has the potential to improve the SNB procedure by facilitating intraoperative visual identification of the sentinel lymph node (SN). The purpose of this study was to evaluate the feasibility of fluorescence tracer imaging for SN detection in conjunction with conventional radio-guided technique. Prospective study of patients with primary OSCC planned for tumor resection and SNB. Thirty patients were injected peritumorally with a bimodal tracer (ICG-99mTc-Nanocoll) followed by lymphoscintigraphy and SPECT/CT to define the SNs and their anatomic allocation preoperatively. SNs were detected intraoperatively with a hand-held gamma-probe and a hand-held NIRF camera. In 29 of 30 subjects (97%), all preoperatively defined SNs could be identified intraoperatively using a combination of radioactive and fluorescence guidance. A total of 94 SNs (mean 3, range 1-5) that were both radioactive and fluorescent ex vivo were harvested. Eleven of 94 SNs (12%) could only be identified in vivo using NIRF imaging, and the majority of those were located in level 1 close to the primary tumor. A combined fluorescent and radioactive tracer for SNB is feasible, and the additional use of NIRF imaging may improve the accuracy of SN identification in oral cancer patients. Intraoperative fluorescence guidance seems of particular value when SNs are located in close proximity to the injection site.


Eradication of Peste des Petits Ruminants Virus and the Wildlife-Livestock Interface.
Growing evidence suggests that multiple wildlife species can be infected with peste des petits ruminants virus (PPRV), with important consequences for the potential maintenance of PPRV in communities of susceptible hosts, and the threat that PPRV may pose to the conservation of wildlife populations and resilience of ecosystems. Significant knowledge gaps in the epidemiology of PPRV across the ruminant community (wildlife and domestic), and the understanding of infection in wildlife and other atypical host species groups (e.g., camelidae, suidae, and bovinae) hinder our ability to apply necessary integrated disease control and management interventions at the wildlife-livestock interface. Similarly, knowledge gaps limit the inclusion of wildlife in the FAO/OIE Global Strategy for the Control and Eradication of PPR, and the framework of activities in the PPR Global Eradication Programme that lays the foundation for eradicating PPR through national and regional efforts. This article reports on the first international meeting on, "Controlling PPR at the livestock-wildlife interface," held in Rome, Italy, March 27-29, 2019. A large group representing national and international institutions discussed recent advances in our understanding of PPRV in wildlife, identified knowledge gaps and research priorities, and formulated recommendations. The need for a better understanding of PPRV epidemiology at the wildlife-livestock interface to support the integration of wildlife into PPR eradication efforts was highlighted by meeting participants along with the reminder that PPR eradication and wildlife conservation need not be viewed as competing priorities, but instead constitute two requisites of healthy socio-ecological systems.


Hierarchical control using networks trained with higher-level forward models.
We propose and develop a hierarchical approach to network control of complex tasks. In this approach, a low-level controller directs the activity of a "plant," the system that performs the task. However, the low-level controller may be able to solve only fairly simple problems involving the plant. To accomplish more complex tasks, we introduce a higher-level controller that controls the lower-level controller. We use this system to direct an articulated truck to a specified location through an environment filled with static or moving obstacles. The final system consists of networks that have memorized associations between the sensory data they receive and the commands they issue. These networks are trained on a set of optimal associations generated by minimizing cost functions. Cost function minimization requires predicting the consequences of sequences of commands, which is achieved by constructing forward models, including a model of the lower-level controller. The forward models and cost minimization are used only during training, allowing the trained networks to respond rapidly. In general, the hierarchical approach can be extended to larger numbers of levels, dividing complex tasks into more manageable subtasks. The optimization procedure and the construction of the forward models and controllers can be performed in similar ways at each level of the hierarchy, which allows the system to be modified to perform other tasks or to be extended for more complex tasks without retraining lower-levels.


Utility of preoperative 3 Tesla pelvic phased-array multiparametric magnetic resonance imaging in prediction of extracapsular extension and seminal vesicle invasion of prostate cancer and its impact on surgical margin status: Experience at a Canadian academic tertiary care centre.
To evaluate the utility of 3 Tesla (3T) pelvic phased-array (PPA) multiparametric magnetic resonance imaging (mpMRI) to predict extracapsular extension (ECE) and seminal vesicle invasion (SVI) and its subsequent effect on radical prostatectomy (RP) surgical margin status. A retrospective evaluation was conducted of RP patients who underwent preoperative 3T PPA mpMRI (without endorectal coil) based on clinical probability of adverse pathological features. Frequencies, specificity, sensitivity, positive predictive value (PPV), and negative predictive value (NPV) of mpMRI in predicting the status of ECE and SVI were calculated. Forty-eight consecutive patients were included. Sensitivity, specificity, PPV, and NPV for 3T PPA mpMRI using T2-weighted sequences with diffusion-weighted imaging (DWI) and dynamic contrast enhanced (DCE) imaging to predict ECE was 39%, 56%, 45%, and 50%, respectively, while SVI prediction was 33%, 95%, 50%, and 91%, respectively. Twelve of the 28 cases predicted as being negative for ECE had positive margins, while two of the 20 cases predicted to be positive for ECE had positive margins. Imaging predicted four cases would have SVI, yet two had positive margins, while of the 44 cases predicted as being negative for SVI, four had positive margins. These findings at our centre suggest that the use of 3T PPA mpMRI using T2-weighted sequences with DWI and DCE in predicting pathological ECE and SVI is of questionable benefit. These mpMRI reports may result in closer dissection of neurovascular bundles and subsequent positive surgical margins. Caution should be exercised when basing intraoperative decisions on mpMRI findings.


Ipl1p-dependent phosphorylation of Mad3p is required for the spindle checkpoint response to lack of tension at kinetochores.
The spindle checkpoint delays anaphase onset until all chromosomes are correctly attached to microtubules. Ipl1 protein kinase (Aurora B) is required to correct inappropriate kinetochore-microtubule attachments and for the response to lack of tension between sister kinetochores. Here we identify residues in the checkpoint protein Mad3p that are phosphorylated by Ipl1p. When phosphorylation of Mad3p at two sites is prevented, the cell's response to reduced kinetochore tension is dramatically curtailed. Our data provide strong evidence for a distinct checkpoint pathway responding to lack of sister kinetochore tension, in which Ipl1p-dependent phosphorylation of Mad3p is a key step.


Differential distribution of three Ca(2+)-activated K(+) channel subunits, SK1, SK2, and SK3, in the adult rat central nervous system.
Ca(2+)-activated, voltage-independent K(+) channels are present in most neurons and mediate the afterhyperpolarizations (AHPs) following action potentials. They present distinct physiological and pharmacological properties and play an important role in controlling neuronal firing frequency and spike frequency adaptation. We used in situ hybridization to characterize the distribution patterns of the three cloned SK channel subunits (SK1-3), the prime candidates likely to underlie Ca(2+)-dependent AHPs in the central nervous system. We found high levels of expression in regions presenting prominent AHP currents, such as, for example, neocortex and CA1-3 layers of the hippocampus (SK1 and SK2), reticularis thalami (SK1 and SK2), supraoptic nucleus (SK3), and inferior olivary nucleus (SK2 and SK3). Our results reveal the functional role of SK channels with defined subunit compositions in some neurons and open the way to the identification of the molecular determinants of AHP currents in many brain regions.


Rhombohedral to Cubic Conversion of GeTe via MnTe Alloying Leads to Ultralow Thermal Conductivity, Electronic Band Convergence, and High Thermoelectric Performance.
In this study, a series of Ge1-xMnxTe (x = 0-0.21) compounds were prepared by a melting-quenching-annealing process combined with spark plasma sintering (SPS). The effect of alloying MnTe into GeTe on the structure and thermoelectric properties of Ge1-xMnxTe is profound. With increasing content of MnTe, the structure of the Ge1-xMnxTe compounds gradually changes from rhombohedral to cubic, and the known R3m to Fm-3m phase transition temperature of GeTe moves from 700 K closer to room temperature. First-principles density functional theory calculations show that alloying MnTe into GeTe decreases the energy difference between the light and heavy valence bands in both the R3m and Fm-3m structures, enhancing a multiband character of the valence band edge that increases the hole carrier effective mass. The effect of this band convergence is a significant enhancement in the carrier effective mass from 1.44 m0 (GeTe) to 6.15 m0 (Ge0.85Mn0.15Te). In addition, alloying with MnTe decreases the phonon relaxation time by enhancing alloy scattering, reduces the phonon velocity, and increases Ge vacancies all of which result in an ultralow lattice thermal conductivity of 0.13 W m-1 K-1 at 823 K. Subsequent doping of the Ge0.9Mn0.1Te compositions with Sb lowers the typical very high hole carrier concentration and brings it closer to its optimal value enhancing the power factor, which combined with the ultralow thermal conductivity yields a maximum ZT value of 1.61 at 823 K (for Ge0.86Mn0.10Sb0.04Te). The average ZT value of the compound over the temperature range 400-800 K is 1.09, making it the best GeTe-based thermoelectric material.


Direct high-temperature ohmic heating of metals as liquid pipes.
When a sufficiently high electric current is passed through a liquid metal, the electromagnetic pressure pinches off the liquid metal and interrupts the flow of current. For the first time the pinch effect has been overcome by use of centrifugal acceleration. By rotation of a pipe of liquid metal, tin or bismuth or their alloys, at sufficiently high speed, it can be heated electrically without intermission of the electric current. One may now heat liquid metallic substances, by resistive (ohmic) heating, to 5000 degrees K and perhaps higher temperatures.


Rnx deficiency results in congenital central hypoventilation.
The genes Tlx1 (Hox11), Enx (Hox11L2, Tlx-2) and Rnx (Hox11L2, Tlx-3) constitute a family of orphan homeobox genes. In situ hybridization has revealed considerable overlap in their expression within the nervous system, but Rnx is singularly expressed in the developing dorsal and ventral region of the medulla oblongata. Tlx1-deficient and Enx-deficient mice display phenotypes in tissues where the mutated gene is singularly expressed, resulting in asplenogenesis and hyperganglionic megacolon, respectively. To determine the developmental role of Rnx, we disrupted the locus in mouse embryonic stem (ES) cells. Rnx deficient mice developed to term, but all died within 24 hours after birth from a central respiratory failure. The electromyographic activity of intercostal muscles coupled with the C4 ventral root activity assessed in a medulla-spinal cord preparation revealed a high respiratory rate with short inspiratory duration and frequent apnea. Furthermore, a coordinate pattern existed between the abnormal activity of inspiratory neurons in the ventrolateral medulla and C4 motorneuron output, indicating a central respiratory defect in Rnx mice. Thus, Rnx is critical for the development of the ventral medullary respiratory centre and its deficiency results in a syndrome resembling congenital central hypoventilation.


Enzymatic detoxification of phosphoramide mustard by soluble fractions from rat organ tissues.
Enzymatic degradation of phosphoramide mustard (PM), the ultimate cytotoxic metabolite of cyclophosphamide (CP), by the cleavage of the phosphorous-nitrogen (P-N) bond was investigated in vitro using 65,000g-soluble fractions from rat organ tissues. In the presence of physiologic bicarbonate in vivo, the P-N bond cleavage of PM was previously found to form 3-(2-chloroethyl)-1,3-oxazolidin-2-one (CNM), which is devoid of antitumor activity. This product was thus quantitated in the present studies using GC/MS and a deuterium-labeled analog (CNM-d8) as the internal standard. Under the experimental conditions, CNM was found to be enzymatically produced from PM in soluble fractions of rat liver, kidney, spleen, and intestine. Mean KM and Vmax values in soluble fractions of rat liver at pH 6.7 and 37 degrees C were determined to be 1.65 +/- 0.536 mM (N = 7) and 6.38 +/- 1.37 microM/min (N = 6), respectively. The enzymic activity of the rat liver soluble fraction was significantly reduced following boiling at 100 degrees C for 5 min. No CNM production was detected from PM incubated in plasma. The P-N bond cleavage for CP and for two other metabolites, 4-ketocyclophosphamide and alcophosphamide, was also investigated using soluble fractions from rat liver similar to that for PM. None of these compounds has been found to form CNM, however, indicating enzyme specificity for the P-N bond in PM. This enzyme probably resembles the previously described phosphamidases.(ABSTRACT TRUNCATED AT 250 WORDS)


VTE, thrombophilia, antithrombotic therapy, and pregnancy: Antithrombotic Therapy and Prevention of Thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines.
The use of anticoagulant therapy during pregnancy is challenging because of the potential for both fetal and maternal complications. This guideline focuses on the management of VTE and thrombophilia as well as the use of antithrombotic agents during pregnancy. The methods of this guideline follow the Methodology for the Development of Antithrombotic Therapy and Prevention of Thrombosis Guidelines: Antithrombotic Therapy and Prevention of Thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines in this supplement. We recommend low-molecular-weight heparin for the prevention and treatment of VTE in pregnant women instead of unfractionated heparin (Grade 1B). For pregnant women with acute VTE, we suggest that anticoagulants be continued for at least 6 weeks postpartum (for a minimum duration of therapy of 3 months) compared with shorter durations of treatment (Grade 2C). For women who fulfill the laboratory criteria for antiphospholipid antibody (APLA) syndrome and meet the clinical APLA criteria based on a history of three or more pregnancy losses, we recommend antepartum administration of prophylactic or intermediate-dose unfractionated heparin or prophylactic low-molecular-weight heparin combined with low-dose aspirin (75-100 mg/d) over no treatment (Grade 1B). For women with inherited thrombophilia and a history of pregnancy complications, we suggest not to use antithrombotic prophylaxis (Grade 2C). For women with two or more miscarriages but without APLA or thrombophilia, we recommend against antithrombotic prophylaxis (Grade 1B). Most recommendations in this guideline are based on observational studies and extrapolation from other populations. There is an urgent need for appropriately designed studies in this population.


Safety evaluation of nivolumab added concurrently to radiotherapy in a standard first line chemo-radiotherapy regimen in stage III non-small cell lung cancer-The ETOP NICOLAS trial.
Chemo-radiotherapy (CRT) and concurrent PD-1 inhibition has shown promising results in pre-clinical models. So far, the feasibility of delivering concurrent CRT and PD-1/PD-L1 inhibition has never been assessed in a clinical trial. NICOLAS is a phase-II trial evaluating the safety and efficacy of nivolumab combined with CRT in stage III NSCLC. Patients received 3 cycles of platinum-based chemotherapy and concurrent RT (66 Gy/33fractions). Nivolumab started concurrently with RT. The primary endpoint was 6-month post-RT rate of grade-≥3-pneumonitis. A formal interim safety analysis (IA) was scheduled when the first 21 patients reached 3 months follow-up post-RT. An early positive safety conclusion would be reached at IA if there were no grade ≥3-pneumonitis in those patients. Efficacy evaluation was planned provided the safety conclusion was reached. As of 13 December 2018, 82 patients were recruited with median follow-up of 13.4 months. The most frequent adverse events (AEs) were anaemia, fatigue and pneumonitis. No unexpected AEs or increased toxicities were observed. For the first 21 patients, no grade-≥3-pneumonitis was observed by the end of the 3-month post-RT follow-up period. The early safety IA provides evidence that the addition of nivolumab to concurrent CRT is safe and tolerable regarding the 6-month rate of pneumonitis grade ≥3 at the one-sided significance level of 5%. Following that, the 1-year progression-free survival will be evaluated in an expanded patient cohort. NICOLAS trial creates the opportunity for assessing the activity of the combination of checkpoint with concurrent CRT in larger prospective trials for locally advanced NSCLC.


Polyphenolic extracts from Olea europea L. protect against cytokine-induced β-cell damage through maintenance of redox homeostasis.
Various pancreatic β-cell stressors, including cytokines, are known to induce oxidative stress, resulting in apoptotic/necrotic cell death and inhibition of insulin secretion. Traditionally, olive leaves or fruits are used for treating diabetes, but the cellular mechanism(s) of their effects are not known. We examined the effects of Olea europea L. (olive) leaf and fruit extracts and their component oleuropein on cytokine-induced β-cell toxicity. INS-1, an insulin-producing β-cell line, was preincubated with or without increasing concentrations of olive leaf or fruit extract or oleuropein for 24 hr followed by exposure to a cytokine cocktail containing 0.15 ng/mL interleukin-1β (IL-1β), 1 ng/mL interferon-γ (IFN-γ), and 1 ng/mL tumor necrosis factor-α (TNF-α) for 6 hr. The cytotoxicity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) testing. Apoptosis was quantified by detecting acridine orange/ethidium bromide-stained condensed nuclei under a fluorescent microscope. The cells exposed to cytokines had a higher apoptotic rate, a decreased viability (MTT), and an increased caspase 3/7 activity. Both extracts and oleuropein partially increased the proportion of living cells and improved the viability of cells after cytokines. The protective effects of extracts on live cell viability were mediated through the suppression of caspase 3/7 activity. Oleuropein did not decrease the amount of both apoptotic and necrotic cells, whereas extracts significantly protected cells against cytokine-induced death. Cytokines led to an increase in reactive oxygen species (ROS) generation and inhibited glutathione level, superoxide dismutase activity, and insulin secretion in INS-1. Insulin secretion was almost completely protected by leaf extract, but was partially affected by fruit extract or oleuropein. Neither cytokines nor olive derivatives had a significant effect on cellular cytochrome c release and catalase activity. Moreover, the cells incubated with each extract or oleuropein showed a significant reduction in cytokine-induced ROS production and ameliorated abnormal antioxidant defense. The molecular mechanism by which olive polyphenols inhibit cytokine-mediated β-cell toxicity appears to be involving the maintenance of redox homeostasis.


CARM1 is an important determinant of ERα-dependent breast cancer cell differentiation and proliferation in breast cancer cells.
Breast cancers with estrogen receptor α (ERα) expression are often more differentiated histologically than ERα-negative tumors, but the reasons for this difference are poorly understood. One possible explanation is that transcriptional cofactors associated with ERα determine the expression of genes which promote a more differentiated phenotype. In this study, we identify one such cofactor as coactivator-associated arginine methyltransferase 1 (CARM1), a unique coactivator of ERα that can simultaneously block cell proliferation and induce differentiation through global regulation of ERα-regulated genes. CARM1 was evidenced as an ERα coactivator in cell-based assays, gene expression microarrays, and mouse xenograft models. In human breast tumors, CARM1 expression positively correlated with ERα levels in ER-positive tumors but was inversely correlated with tumor grade. Our findings suggest that coexpression of CARM1 and ERα may provide a better biomarker of well-differentiated breast cancer. Furthermore, our findings define an important functional role of this histone arginine methyltransferase in reprogramming ERα-regulated cellular processes, implicating CARM1 as a putative epigenetic target in ER-positive breast cancers.


Characteristics of the single-longitudinal-mode planar-waveguide external cavity diode laser at 1064 nm.
We describe the characteristics of the planar-waveguide external cavity diode laser (PW-ECL). To the best of our knowledge, it is the first butterfly-packaged 1064 nm semiconductor laser that is stable enough to be locked to an external frequency reference. We evaluated its performance from the viewpoint of precision experiments. Using a hyperfine absorption line of iodine, we suppressed its frequency noise by a factor of up to 10(4) at 10 mHz. The PW-ECL's compactness and low cost make it a candidate to replace traditional Nd:YAG nonplanar ring oscillators and fiber lasers in applications that require a single longitudinal mode.


Measuring decision weights in recognition experiments with multiple response alternatives: comparing the correlation and multinomial-logistic-regression methods.
Psychophysical "reverse-correlation" methods allow researchers to gain insight into the perceptual representations and decision weighting strategies of individual subjects in perceptual tasks. Although these methods have gained momentum, until recently their development was limited to experiments involving only two response categories. Recently, two approaches for estimating decision weights in m-alternative experiments have been put forward. One approach extends the two-category correlation method to m > 2 alternatives; the second uses multinomial logistic regression (MLR). In this article, the relative merits of the two methods are discussed, and the issues of convergence and statistical efficiency of the methods are evaluated quantitatively using Monte Carlo simulations. The results indicate that, for a range of values of the number of trials, the estimated weighting patterns are closer to their asymptotic values for the correlation method than for the MLR method. Moreover, for the MLR method, weight estimates for different stimulus components can exhibit strong correlations, making the analysis and interpretation of measured weighting patterns less straightforward than for the correlation method. These and other advantages of the correlation method, which include computational simplicity and a close relationship to other well-established psychophysical reverse-correlation methods, make it an attractive tool to uncover decision strategies in m-alternative experiments.


Negative/zero thermal expansion in black phosphorus nanotubes.
Black phosphorus nanotubes (BPNTs) as new 1D materials have very promising prospects in nanoelectronic devices. Taking into account that the effect of finite temperature on nanoelectronic devices is indispensable, we perform a comprehensive first-principles study of thermal expansion of BPNTs. It is surprisingly found that the thermal expansion can be tuned from positive thermal expansion (PTE) to zero thermal expansion (ZTE) and even to negative thermal expansion (NTE) by changing the diameter and bonding in the unique configurations of armchair and zigzag BPNTs. The thermal expansion of zigzag BPNTs is diameter independent, in contrast, the size effect of thermal expansion is evidently presented in the armchair BPNTs. Accordingly, it is more manageable to adjust the coefficient of thermal expansion (CTE) in the armchair BPNTs. Unusual phonon hardening with the increase of temperature can be detected in both types of nanotubes. The unique dynamical modes and different vibrational amplitudes are analyzed. Underlying physical mechanisms have been explored. This work can provide some inspiration for exploring the abnormal thermal expansion in nanotubes and how to regulate these properties.


Receptors for IgG: subclass specificity of receptors on different mouse cell types and the definition of two distinct receptors on a macrophage cell line.
To evaluate subclass specificity and aggregate size requirements of IgG receptors on mouse cells, we measured binding of radiolabeled monomeric and BDB-aggregated mouse myeloma proteins fractionated into various sizes by means of gel filtration. Monomers, tetramers, and high molecular weight (approximately 10(7) daltons) aggregates were used. The various cells and cell lines studied could be segregated into three patterns of reactivity: (a) Macrophage and macrophage-like cell lines bound monomer IgG2a preferentially; high molecular weight IgG aggregates bound as follows: IgG1 = IgG2b = IgG2a. (b) Lymphoid lines D2N and S49 showed no capacity to bind monomer IgG2a; high molecular weight aggregates bound as follows: IgG1 = IgG2b less than IgG2a. (c) Other Thy-1-positive lymphoid cell lines (EL4 and L5178) and normal T and B cells showed no capacity to bind monomer IgG; high molecular weight IgG aggregates bound to a lesser extent than to cells of the first two categories in the following manner: IgG1 less than IgG2b greater than or equal to IgG2a. The variable pattern of reactivity of the macrophage-like cell lines with monomer and aggregated IgG suggested that two distinct receptors for IgG were present: one capable of binding IgG2a and another capable of binding all aggregates. Further evidence for this hypothesis was obtained by analysis of the inhibitory capacity of different IgG subclasses on the binding of aggregated IgG and monomer IgG2a to P388 cells. Inhibition of monomer IgG2a binding was effected only by monomer or aggregated IgG2a, whereas inhibition of binding of aggregated IgG1 or IgG2b was noted with aggregates of all three subclasses with some preferential inhibition by monomer IgG2b being observed. Furthermore, monomer IgG2b binding was preferentially inhibitable by monomer IgG2b. It is postulated from these data that two receptor sites are present on this macrophage-like cell line, one reactive with aggregates of all three subclasses as well as monomer IgG2b, and another receptor specific for monomer IgG2a which also binds aggregated IgG2a. Support of this concept was obtained by trypsinization experiments in which the binding of monomer IgG2a was markedly decreased by trypsin treatment of cells, whereas the binding of aggregated IgG2b was unaffected by this treatment.