Full-length recombinant Plasmodium falciparum VAR2CSA binds specifically to CSPG and induces potent parasite adhesion-blocking antibodies.
Plasmodium falciparum malaria remains one of the world's leading causes of human suffering and poverty. Each year, the disease takes 1-3 million lives, mainly in sub-Saharan Africa. The adhesion of infected erythrocytes (IEs) to vascular endothelium or placenta is the key event in the pathogenesis of severe P. falciparum infection. In pregnant women, the parasites express a single and unique member of the P. falciparum erythrocyte membrane protein 1 (PfEMP1) family named VAR2CSA, which is associated with the ability of the IEs to adhere specifically to chondroitin sulphate A (CSA) in the placenta. Several Duffy-binding-like domains from VAR2CSA molecules have been shown in vitro to bind to CSA, but it has also been demonstrated that Duffy-binding-like domains from PfEMP1 proteins other than VAR2CSA can bind CSA. In addition, the specificity of the binding of VAR2CSA domains to glycosaminoglycans does not match that of VAR2CSA-expressing IEs. This has led to speculation that the domains of native VAR2CSA need to come together to form a specific binding site or that VAR2CSA might bind to CSA through a bridging molecule. Here, we describe the expression and purification of the complete extracellular region of VAR2CSA secreted at high yields from insect cells. Using surface plasmon resonance, we demonstrate that VAR2CSA alone binds with nanomolar affinity to human chondroitin sulphate proteoglycan and with significantly weaker affinity to other glycosaminoglycans, showing a specificity similar to that observed for IEs. Antibodies raised against full-length VAR2CSA completely inhibit recombinant VAR2CSA binding, as well as parasite binding to chondroitin sulphate proteoglycan. This is the first study to describe the successful production and functionality of a full-length PfEMP1. The specificity of the binding and anti-adhesion potency of induced IgG, together with high-yield production, encourages the use of full-length PfEMP1 in vaccine development strategies.


Nuclear factor-kappa B subunits and their prognostic cancer-specific survival value in renal cell carcinoma patients.
Better characterisation and understanding of renal cell carcinoma (RCC) development and progression lead to better diagnosis and clinical outcomes. In this study, expression of nuclear factor-kappa B (NF-κB) subunits: p65 (RelA), p105/p50, p100/p52, and cRel in RCC tissue were compared with corresponding normal kidney, along with tumour characteristics and survival outcome. Ninety-six cases of RCC with paired normal kidney were analysed. Clinicopathological data, demographics and survival data were available. Immunohistochemistry (IHC) for NF-κB subtypes was analysed using the Aperio digital pathology system for overall cellular expression and localisation. The prognostic cancer-specific survival value of the subunits in RCC patients was analysed. Approximately 50% of patients had clinical stage T1, with 22 patients having metastases at presentation. RCC subtypes were: clear cell (n = 76); papillary (n = 11); chromophobe (n = 5); clear cell tubulopapillary (n = 3); and one multilocular cystic RCC. Median follow up was 54.5 months (0.2-135), with 28 deaths at time of analysis. NF-κB p65 had higher overall and nuclear expressions, with lower overall and nuclear expressions of p50, p52 and cRel in RCC compared with normal kidney. Higher expressions of p65 (nuclear), p52 (overall and nuclear) and p50 (overall) correlated significantly with worse cancer-specific survival. This is the first large series of analysis of expression of NF-κB subunits in RCC. Especially with regards to the less studied subunits (p52, p50, cRel), our results allow a better understanding the role of NF-κB in RCC development and progression, and may pave the way for future targeted NF-κB subunit specific therapies.


Microarray comparative genomic hybridization analysis of tubular breast carcinoma shows recurrent loss of the CDH13 locus on 16q.
Tubular breast carcinoma is a highly differentiated carcinoma with an excellent prognosis. Distinct genetic alterations in tubular breast carcinoma cells have been described, especially broad genetic losses on the q-arm of chromosome 16. These are more common in lobular breast carcinoma and low-grade ductal carcinoma in situ than in ductal breast carcinoma and high-grade ductal carcinoma in situ. To further delineate the molecular changes involved in tubular breast carcinoma more precisely, we examined 23 formalin-fixed and paraffin wax-embedded tissue samples (21 of tubular breast carcinoma and 2 of nonneoplastic breast epithelium) by microarray-based comparative genomic hybridization focusing on 287 genomic target clones of oncogenes and tumor suppressor genes. The results obtained from all nonneoplastic tissue samples of breast epithelium indicate no DNA copy number changes. In the tubular breast carcinoma samples, the highest frequencies for DNA sequence copy number losses were detected for CDH13 (in 86% of the samples) and MSH2, KCNK12 (in 52% of the samples). The highest frequencies of DNA sequence copy number gains were detected for HRAS and D13S319XYZ (each in 62% of the samples). Using principal component analysis, 3 subgroups of tubular breast carcinomas showing relative genetic changes were identified. For validation, the most frequent DNA copy number loss for CDH13 (18/21) was confirmed using fluorescence in situ hybridization in 4 of 5 tubular breast carcinomas analyzed. The newly identified genes with considerable copy number changes may include so far unknown candidate genes for the development and progression of tubular breast carcinoma, such as CDH13. The study provides the starting point for further delineating their detailed influence on the pathogenesis of tubular breast carcinoma.


Effect of Flooding and the nosZ Gene in Bradyrhizobia on Bradyrhizobial Community Structure in the Soil.
We investigated the effects of the water status (flooded or non-flooded) and presence of the nosZ gene in bradyrhizobia on the bradyrhizobial community structure in a factorial experiment that examined three temperature levels (20°C, 25°C, and 30°C) and two soil types (andosol and gray lowland soil) using microcosm incubations. All microcosms were inoculated with Bradyrhizobium japonicum USDA6T, B. japonicum USDA123, and B. elkanii USDA76T, which do not possess the nosZ gene, and then half received B. diazoefficiens USDA110Twt (wt for the wild-type) and the other half received B. diazoefficiens USDA110ΔnosZ. USDA110Twt possesses the nosZ gene, which encodes N2O reductase; 110ΔnosZ, a mutant variant, does not. Changes in the community structure after 30- and 60-d incubations were investigated by denaturing-gradient gel electrophoresis and an image analysis. USDA6T and 76T strains slightly increased in non-flooded soil regardless of which USDA110T strain was present. In flooded microcosms with the USDA110Twt strain, USDA110Twt became dominant, whereas in microcosms with the USDA110ΔnosZ, a similar change in the community structure occurred to that in non-flooded microcosms. These results suggest that possession of the nosZ gene confers a competitive advantage to B. diazoefficiens USDA110T in flooded soil. We herein demonstrated that the dominance of B. diazoefficiens USDA110Twt within the soil bradyrhizobial population may be enhanced by periods of flooding or waterlogging systems such as paddy-soybean rotations because it appears to have the ability to thrive in moderately anaerobic soil.


Carotid artery stenting vs carotid endarterectomy: meta-analysis and diversity-adjusted trial sequential analysis of randomized trials.
The role of carotid artery stenting (CAS) when compared with carotid endarterectomy (CEA) is controversial, with recent trials showing an increased risk of harm with CAS. To evaluate the periprocedural and intermediate to long-term benefits and harms of CAS compared with CEA. PubMed, EMBASE, and Cochrane Central Register of Controlled Trials searches for randomized clinical trials until June 2010 of CAS compared with CEA for carotid artery disease. Periprocedural (≤30-day) outcomes (death, myocardial infarction [MI], or stroke; death or any stroke; any stroke; and MI) and intermediate to long-term outcomes (outcomes as in the Stenting and Angioplasty With Protection in Patients at High Risk for Endarterectomy [SAPPHIRE] trial: composite of periprocedural death, MI, or stroke plus ipsilateral stroke or death thereafter; periprocedural death or stroke plus ipsilateral stroke thereafter; death or any stroke; and any stroke) were evaluated. Two of us independently extracted data in duplicate. Baseline characteristics, inclusion and exclusion criteria, use of an embolic protection device, US vs non-US study, and the earlier-mentioned outcomes of interest were extracted from each trial. We identified 13 randomized clinical trials randomizing 7477 participants. Carotid artery stenting was associated with an increased risk of periprocedural outcomes of death, MI, or stroke (odds ratio = 1.31; 95% confidence interval, 1.08-1.59), 65% and 67% increases in death or stroke and any stroke, respectively, but with 55% and 85% reductions in the risk of MI and cranial nerve injury, respectively, when compared with CEA. The trial sequential monitoring boundary was crossed by the cumulative z curve, suggesting firm evidence for at least a 20% relative risk increase of periprocedural death or stroke and any stroke and at least a 15% reduction in MI with CAS compared with CEA. Similarly, CAS was associated with 19%, 38%, 24%, and 48% increases in the intermediate to long-term outcomes of SAPPHIRE-like outcome, periprocedural death or stroke and ipsilateral stroke thereafter, death or any stroke, and any stroke, respectively. The trial sequential monitoring boundary was crossed by the cumulative z curve, suggesting firm evidence for at least a 20% relative risk increase of any stroke. In this largest and most comprehensive meta-analysis to date using outcomes that are standard in contemporary studies, CAS was associated with an increased risk of both periprocedural and intermediate to long-term outcomes, but with a reduction in periprocedural MI and cranial nerve injury. Strategies are urgently needed to identify patients who are best served by CAS vs CEA.


Genome-wide identification of inter-individually variable DNA methylation sites improves the efficacy of epigenetic association studies.
Epigenome-wide association studies, which searches for blood-based DNA methylation signatures associated with environmental exposures and/or disease susceptibilities, is a promising approach to a better understanding of the molecular aetiology of common diseases. To carry out large-scale epigenome-wide association studies while avoiding false negative detection, an efficient strategy to determine target CpG sites for microarray-based or sequencing-based DNA methylation profiling is essentially needed. Here, we propose and validate a hypothesis that a strategy focusing on CpG sites with high DNA methylation level variability may attain an improved efficacy. Through whole-genome bisulfite sequencing of purified blood cells collected from > 100 apparently healthy subjects, we identified ~2.0 million inter-individually variable CpG sites as potential targets. The efficacy of our strategy was estimated to be 3.7-fold higher than that of the most frequently used strategy. Our catalogue of inter-individually variable CpG sites will accelerate the discovery of clinically relevant DNA methylation biomarkers in future epigenome-wide association studies.


A fluorescence polarization assay for the experimental validation of an in silico model of the chemokine CXCL8 binding to receptor-derived peptides.
Peptide based inhibitors of protein-protein interactions are of great interest in proteomics, structural biology and medicinal chemistry. Optimized inhibitors can be designed by experimental approaches or by computational prediction. Ideally, computational models are adjusted to the peptide-protein complex of interest according to experimental data obtained in specific binding experiments. The chemokine CXCL8 (interleukin-8) is an interesting target for drug discovery due to its role in inflammatory diseases. Given the available structural data and information on its receptor interactions it constitutes a basis for the rational design of inhibitor peptides. Starting from the reported structure of CXCL8 in complex with a peptide derived from its receptor CXCR1 we developed a computational docking procedure to estimate the changes in binding energy as a function of individual amino acid exchanges. This indicates whether the respective amino acid residue must be preserved or can be substituted to maintain or improve affinity, respectively. To validate and improve the assumptions made in this docking simulation we established a fluorescence polarization assay for receptor-derived peptides binding to CXCL8. A peptide library was tested comprising selected mutants characterized by docking simulations. A number of predictions regarding electrostatic interactions were confirmed by these experiments and it was revealed that the model needed to be corrected for backbone flexibility. Therefore, the assay presented here is a promising tool to systematically improve the computational model by iterative cycles of modeling, experimental validation and refinement of the algorithm, leading to a more reliable model and peptides with improved affinity.


Silica dioxide colloidal solutions is efficient in the treatment of chronic periodontitis: a case control study.
The objective of this study was to compare the efficacy of supportive periodontal therapy (i.e. scaling and rooth planning, SRP) alone versus a chemical device silica dioxide (SiO2) colloidal solutions (SDCS) used in association with SRP in the treatment of chronic periodontitis in adult patients. A total of 20 patients with a diagnosis of chronic periodontitis (40 localized chronic periodontitis sites) in the age group of 35 to 55 were selected. None of these patients have previously received any surgical or non-surgical periodontal therapy and demonstrated radiographic evidence of moderate bone loss. Two non-adjacent sites in separate quadrants were selected in each patient to monitorize treatment efficacy (split mouth design). Clinical pocket depth (PD) and microbial analysis (MA) were analyzed at baseline and 15th day. SPSS program and paired simple statistic T-test were used to detect significant differences. Total bacteria loading, Tannerella Forsitia and Treponema Denticola loading were statistically reduced when SiO2 is locally delivered. SDCS gel is an adjuvant therapy which should be added to SRP in the management of moderate to severe chronic periodontitis.


Patterns of theta oscillation reflect the neural basis of individual differences in epistemic motivation.
Theta oscillations in the EEG have been shown to reflect ongoing cognitive processes related to mental effort. Here, we show that the pattern of theta oscillation in response to varying cognitive demands reflects stable individual differences in the personality trait epistemic motivation: Individuals with high levels of epistemic motivation recruit relatively more cognitive resources in response to situations possessing high, compared to low, cognitive demand; individuals with low levels do not show such a specific response. Our results provide direct evidence for the theory of the construct need for cognition and add to our understanding of the neural processes underlying theta oscillations. More generally, we provide an explanation how individual differences in personality traits might be represented on a neural level.


Polarization mode coupling in circularly birefringent gratings.
Polarization mode coupling in circularly birefringent gratings is analyzed. It is numerically found that efficient LP01x-LP02y mode coupling (where LP is linear polarization) is possible in a 50-cm-long circularly birefringent fiber grating formed in a terbium-doped borosilicate glass fiber and that complete LP01x-LP02y and LP01x-LP03y mode couplings result after a few-centimeter-long circularly birefringent grating that is formed in a bismuth-substitute iron garnet waveguide. Various parameters of polarization mode coupling in a number of circularly birefringent gratings are also computed.


S-Nitrosothiols (SNO) as light-responsive molecular activators for post-synthesis fluorescence augmentation in fluorophore-loaded nanospheres.
Dye-loaded, fluorescent nanoparticles (FNPs) have been extensively studied as promising imaging or probing agents for therapeutic and biomedical applications, because of improved photostability and reduced cytotoxicity (compared with free dyes). The synthesis of FNPs often involves entrapment of fluorescent dye molecules into nanostructures, which, however, would encounter a problem associated with the formation of molecular aggregates within the confined matrix space. The formation of nonfluorescent aggregates seems unavoidable for some conventional fluorescent dyes, thereby leading to fluorescent quenching. The problem is well-recognized, but frequently ignored; and FNPs are usually applied as prepared without addressing it either during or after the synthesis of FNPs. The ignorance may be due to the difficulty in altering post-synthetically the intraparticulate molecular arrangements and interactions. Herein, we describe how light-responsive S-nitrosothiol (SNO) can be engineered into fluorophore-loaded silica nanoparticles for efficient dye-loading and post-synthesis fluorescence augmentation. Silica nanoparticles loaded with various fluorescent molecules were prepared using one-pot, simultaneous, acid-catalyzed sol-gel condensation and nitrosation of a single mercaptosilane source, 3-mercaptopropyl trimethoxysilane (MPTMS), followed by nanoprecipitation. We first show how doxorubicin-loaded silica NPs respond to stepwise visible-light exposure and exhibit ON/OFF fluorescent response. We then demonstrate that rhodamine 6G (R6G) can be stably incorporated into SNO-enriched silica nanostructure with negligible payload leakage (<0.2%) and ∼1000-fold reduction in cytotoxicity (compared with free R6G). Remarkably, visible light irradiation leads to ∼100-fold increase in fluorescent intensity. Thus, for the first time, SNO is proposed as a light-responsive entity for post-synthesis fluorescence intensification in FNPs.


Examining the range of normal intraindividual variability in neuropsychological test performance.
Neuropsychologists often diagnose cerebral dysfunction based, in part, on marked variation in an individual's cognitive test performance. However, little is known about what constitutes the normal range of intraindividual variation. In this study, after excluding 54 individuals with significant health problems, we derived 32 z-transformed scores from 15 tests administered to 197 adult participants in a study of normal aging. The difference between each person's highest and lowest scores was computed to assess his or her maximum discrepancy (MD). The resulting MD values ranged from 1.6 to 6.1 meaning that the smallest MD shown by any person was 1.6 standard deviations (SDs) and the largest MD shown by any person was 6.1 SDs. Sixty-six percent of participants produced MD values that exceeded 3 SDs. Eliminating each person's highest and lowest test scores decreased their MDs, but 27% of the participants still produced MD values exceeding 3. Although MD values appeared to increase with age, adjusting test scores for age, which is standard in clinical practice, did not correct for this. These data reveal that marked intraindividual variability is very common in normal adults, and underscore the need to base diagnostic inferences on clinically recognizable patterns rather than psychometric variability alone.


Emergence of a ribotype 244 strain of Clostridium difficile associated with severe disease and related to the epidemic ribotype 027 strain.
We identified 12 patients with Clostridium difficile infection between July 2011 and March 2012 from whom an unusual C. difficile strain was isolated. This strain had a single-nucleotide deletion of the tcdC gene at position 117 and binary toxin genes, which are characteristic of the hypervirulent ribotype (RT) 027 strain. A retrospective cohort study of 12 patients infected with C. difficile RT244 and 24 patients infected with non-RT244/non-RT027 strains matched for place of diagnosis and time of collection of specimen was performed. We performed whole-genome sequencing to understand the relationship of the RT244 strain to other C. difficile strains and further understand its virulence potential. Clostridium difficile RT244 was associated with more severe disease and a higher mortality rate. Phylogenomic analysis using core genome single-nucleotide polymorphisms showed that RT244 is in the same genetic clade (clade 2) as RT027 but is distinct from all RT027 strains. The pathogenicity locus of the RT244 strain encodes a variant toxin B, and this was confirmed by demonstration of Clostridium sordellii-like cytopathic effect on Vero cells. Toxin B production in culture supernatants was lower than that seen with a RT027 strain. Our findings demonstrate the pathogenic potential of this RT244 C. difficile strain and emphasize the importance of ongoing surveillance for emergent strains.


A bipedal DNA motor that travels back and forth between two DNA origami tiles.
In this work, the successful operation of a dynamic DNA device constructed from two DNA origami building blocks is reported. The device includes a bipedal walker that strides back and forth between the two origami tiles. Two different DNA origami tiles are first prepared separately; they are then joined together in a controlled manner by a set of DNA strands to form a stable track in high yield as confirmed by single-molecule fluorescence (SMF). Second, a bipedal DNA motor, initially attached to one of the two origami units and operated by sequential interaction with "fuel" and "antifuel" DNA strands, moves from one origami tile to another and then back again. The operational yield, measured by SMF, was similar to that of a motor operating on a similar track embedded in a single origami tile, confirming that the transfer across the junction from one tile to the other does not result in dissociation that is any more than that of steps on a single tile. These results demonstrate that moving parts can reliably travel from one origami unit to another, and it demonstrates the feasibility of dynamic DNA molecular machines that are made of more than a single origami building block. This study is a step toward the development of motors that can stride over micrometer distances.


Use of NMR to study serpin function.
Two-dimensional heteronuclear [1H,15N] single quantum correlation NMR spectra of serpins show dramatic changes between native and loop-inserted conformations, making them very sensitive reporters of the serpin conformation. Much of the spectral overlap that arises when all amides are 15N labelled can be removed by use of selective labelling of a single type of amino acid, such as alanine. The method allows ready determination of whether loop insertion is present, and to what extent, as well as providing information on motional freedom of components of the complex and of the reactive center loop. With label introduced separately into the proteinase, information can also be obtained on the conformational changes brought about in that moiety by complex formation. In addition, with the use of cryoprobes, high field spectrometers, TROSY-based signal detection and deuteration, samples as small as 1-2mg can easily be examined, making it applicable to a wide range of serpins, including those that can only be expressed in mammalian cells.


Developing a disability determination model using a decision support system in Taiwan: a pilot study.
The aims of our study were to: (1) develop the Disability Grading Decision Support System (DGDSS) and to (2) compare the new International Classification of Functioning, Disability, and Health (ICF)-based disability determination tool (ICF-DDT) with the diagnosis-based disability determination tool (D-DDT). A total of 9357 patients recruited from 236 accredited institutions were evaluated using the ICF-DDT and the D-DDT, and the presence, severity, and category of the disability identified using the two determination tools were compared. In the DGDSS, the ICF-DDT consisted of four models comprising nine modules to determine the presence and the severity of the disability. The differences between models (modules) are the different combinations of World Health Organization Disability Assessment Schedule 2.0 (WHODAS 2.0) and Scale of Body Functions and Structures. Compared with the D-DDT, more patients were determined to be disability-free when using the ICF-DDT. Module 1-1 had the highest profoundly severe rate, and module 2-2 had the highest mild and moderate disability rates. Module 2-1 had the highest severe disability rate. Module 1-1 resulted in the smallest difference, and module 3-1 resulted in the largest difference, compared with the D-DDT. Feedback from users suggested that the DGDSS is a robust system if the original data are accurate. The presence, severity, and category of the disability determined using the ICF-DDT and the D-DDT were significantly different. The results of the DGDSS provide information for policymakers to determine the optimal allocation of social welfare and medical resources for people with disabilities.


Berry-phase-induced dimerization in one-dimensional quadrupolar systems.
We investigate the effect of the Berry phase on quadrupoles that occur, for example, in the low-energy description of spin models. Specifically, we study here the one-dimensional bilinear-biquadratic spin-one model. An open question for many years about this model is whether it has a nondimerized fluctuating nematic phase. The dimerization has recently been proposed to be related to Berry phases of the quantum fluctuations. We use an effective low-energy description to calculate the scaling of the dimerization according to this theory and then verify the predictions using large scale density-matrix renormalization group simulations, giving good evidence that the state is dimerized all the way up to its transition into the ferromagnetic phase. We furthermore discuss the multiplet structure found in the entanglement spectrum of the ground state wave functions.


Human prorenin.
Human prorenin is the enzymatically inactive biosynthetic precursor of renin. Recent interest has focused on the posttranslational sorting and processing of prorenin to renin since markedly increased levels of circulating prorenin have been associated with both physiological and pathological changes. These observations raise the question of whether prorenin processing may be a regulatory event in renin production in the kidney. In the juxtaglomerular cells of the kidney, prorenin can be sorted to either of two pathways: 1) the regulated pathway, which is mediated by secretory granules, where a thiol protease resembling cathepsin B processes prorenin to renin by cleavage of the amino terminal 43-amino acid prosegment, which allows exposure of the active site of renin, or 2) the constitutive pathway, which is not regulated and does not involve conversion of prorenin to renin. Studies in which segments of prorenin are modified by site-directed mutagenesis suggest that the prosegment and glycosylation are not required for sorting, although they may influence or participate in sorting, or both. Certain areas in the prosegment are important determinants of enzyme activity and ability to cleave the prosegment. Further structural analysis of prorenin will be useful to assess details of its sorting and processing. In addition, a number of extrarenal tissues such as uterine lining, ovarian theca, corpus luteum, pituitary, and adrenal, express the renin gene. These tissues have different capabilities to sort and process prorenin compared with kidney, and some tissues secrete only prorenin. Whether prorenin-to-renin conversion is necessary to activate these local renin-angiotensin systems is a key issue.(ABSTRACT TRUNCATED AT 250 WORDS)


Elongator subunit 3 (ELP3) modifies ALS through tRNA modification.
Amyotrophic lateral sclerosis (ALS) is a fatal degenerative motor neuron disorder of which the progression is influenced by several disease-modifying factors. Here, we investigated ELP3, a subunit of the elongator complex that modifies tRNA wobble uridines, as one of such ALS disease modifiers. ELP3 attenuated the axonopathy of a mutant SOD1, as well as of a mutant C9orf72 ALS zebrafish model. Furthermore, the expression of ELP3 in the SOD1G93A mouse extended the survival and attenuated the denervation in this model. Depletion of ELP3 in vitro reduced the modified tRNA wobble uridine mcm5s2U and increased abundance of insoluble mutant SOD1, which was reverted by exogenous ELP3 expression. Interestingly, the expression of ELP3 in the motor cortex of ALS patients was reduced and correlated with mcm5s2U levels. Our results demonstrate that ELP3 is a modifier of ALS and suggest a link between tRNA modification and neurodegeneration.


Pharmacodynamics of the antibacterial effect and emergence of resistance to tomopenem, formerly RO4908463/CS-023, in an in vitro pharmacokinetic model of Staphylococcus aureus infection.
The antibacterial effects (ABE) of tomopenem (formerly RO4908463/CS-023) against seven Staphylococcus aureus strains (methicillin-resistant S. aureus [MRSA] strain tomopenem MICs, 0.5 to 16 mg/liter; methicillin-sensitive S. aureus [MSSA] strain tomopenem MIC, 0.06 mg/liter) were studied in an in vitro pharmacokinetic model. Initially, two human doses were simulated, 750 mg every 8 hours (8hly) and 1,500 mg 8hly intravenously, using S. aureus at a standard inoculum of 10(6) CFU/ml. There was a rapid clearance of bacteria from the model by 12 h after drug exposure with most strains. Clearance was not related to the tomopenem MIC. The ABE of these two tomopenem dose regimens were also tested at a high inoculum, 10(8) CFU/ml; in all simulations, there was a >4-log drop in viable count at 24 h. Strains were not cleared from the model at 10(8) CFU/ml, in contrast to what was seen for the standard inoculum. When the ABE of tomopenem at 750 mg 8hly was compared to those of vancomycin, tomopenem was seen to have a superior effect, as measured by the area under the bacterial kill curve at 24 h (AUBKC24) and 48 h (P < 0.05). Dose ranging studies were performed to provide time-above-MIC (T>MIC) drug exposures of 0 to 100% (8 to 10 doses per strain) with five MRSA/MSSA strains. The T>MIC for a 24-h bacteriostatic effect was 8% +/- 5% (range, 1.3% to 15.4%); the T>MIC for a 4-log drop in viable count was 32% +/- 18% (range, 12.8% to 36.2%). The T>MIC for a 90% maximum response using AUBKC24 as ABE was 24.9% +/- 15.7%. Inoculum had little impact on T>MIC exposures for ABE. There was emergence of resistance to tomopenem in the dose ranging studies, with increased growth of subpopulations on plates containing tomopenem at 2x and 4x the MIC compared to what was seen for preexposure population analysis at T>MICs of <20%. The pharmacodynamics of tomopenem against S. aureus is similar to those of other members of the carbapenem class, with the exception that MRSA is included. These data indicate that tomopenem will have clinically useful activity against MRSA at T>MICs achievable in humans.


Lymphatic pump treatment as an adjunct to antibiotics for pneumonia in a rat model.
Lymphatic pump treatment (LPT) is a technique used by osteopathic physicians as an adjunct to antibiotics for patients with respiratory tract infections, and previous studies have demonstrated that LPT reduces bacterial load in the lungs of rats with pneumonia. Currently, it is unknown whether LPT affects drug effcacy. To determine whether the combination of antibiotics and LPT would reduce bacterial load in the lungs of rats with acute pneumonia. Rats were infected intranasally with 5×107 colony-forming units (CFU) of Streptococcus pneumoniae. At 24, 48, and 72 hours after infection, the rats received no therapy (control), 4 minutes of sham therapy, or 4 minutes of LPT, followed by subcutaneous injection of 40 mg/kg of levofoxacin or sterile phosphate-buffered saline. At 48, 72, and 96 hours after infection, the spleens and lungs were collected, and S pneumoniae CFU were enumerated. Blood was analyzed for a complete blood cell count and leukocyte differential count. At 48 and 72 hours after infection, no statistically significant differences in pulmonary CFU were found between control, sham therapy, or LPT when phosphate-buffered saline was administered; however, the reduction in CFU was statistically significant in all rats given levofoxacin. The combination of sham therapy and levofoxacin decreased bacterial load at 72 and 96 hours after infection, and LPT and levofoxacin significantly reduced CFU compared with sham therapy and levofoxacin at both time points (P<.05). Colony-forming units were not detected in the spleens at any time. No statistically significant differences in hematologic findings between any treatment groups were found at any time point measured. The results suggest that 3 applications of LPT induces an additional protective mechanism when combined with levofoxacin and support its use as an adjunctive therapy for the management of pneumonia; however, the mechanism responsible for this protection is unclear.


The imprinted gene Magel2 regulates normal circadian output.
Mammalian circadian rhythms of activity are generated within the suprachiasmatic nucleus (SCN). Transcripts from the imprinted, paternally expressed Magel2 gene, which maps to the chromosomal region associated with Prader-Willi Syndrome (PWS), are highly enriched in the SCN. The Magel2 message is circadianly expressed and peaks during the subjective day. Mice deficient in Magel2 expression entrain to light cycles and express normal running-wheel rhythms, but with markedly reduced amplitude of activity and increased daytime activity. These changes are associated with reductions in food intake and male fertility. Orexin levels and orexin-positive neurons in the lateral hypothalamus are substantially reduced, suggesting that some of the consequences of Magel2 loss are mediated through changes in orexin signaling. The robust rhythmicity of Magel2 expression in the SCN and the altered behavioral rhythmicity of null mice reveal Magel2 to be a clock-controlled circadian output gene whose disruption results in some of the phenotypes characteristic of PWS.


Design, synthesis, biological evaluation, and molecular modeling studies of TIBO-like cyclic sulfones as non-nucleoside HIV-1 reverse transcriptase inhibitors.
TIBO- and TBO-like sulfone derivatives 1 and 2 were designed, synthesized, and tested for their ability to block the replication cycle of HIV-1 in infected cells. The anti-HIV-1 activities of sulfones 3, which were intermediates in the syntheses of 1 and 2, were also evaluated. Surprisingly, the sulfone analogues of TIBO R82913 (compounds 1) were inactive, whereas interesting results were obtained for truncated derivatives 2. Compound 2 w was the most potent among this series in cell-based assays (EC50=0.07 microM, CC50>200 microM, SI>2857). It was twofold less potent than R82913, but more selective. An X-ray crystallographic analysis was carried out to establish the absolute configuration of 2 w and its enantiomer 2 x, which were obtained by semipreparative HPLC of 2 v, one of the most potent racemates. Compounds 1-3 were proven to target HIV-1 RT. In fact, representative derivatives inhibited recombinant HIV-1 RT in vitro at concentrations similar to those active in cell-based assays. 3D QSAR studies and docking simulations were developed on TIBO- and TBO-like sulfone derivatives to rationalize their anti-HIV-1 potencies and to predict the activity of novel untested sulfone derivatives. Predictive 3D QSAR models were obtained with a receptor-based alignment by docking of TIBO- and TBO-like derivatives into the NNBS of RT.


Integrated model of multiple kernel learning and differential evolution for EUR/USD trading.
Currency trading is an important area for individual investors, government policy decisions, and organization investments. In this study, we propose a hybrid approach referred to as MKL-DE, which combines multiple kernel learning (MKL) with differential evolution (DE) for trading a currency pair. MKL is used to learn a model that predicts changes in the target currency pair, whereas DE is used to generate the buy and sell signals for the target currency pair based on the relative strength index (RSI), while it is also combined with MKL as a trading signal. The new hybrid implementation is applied to EUR/USD trading, which is the most traded foreign exchange (FX) currency pair. MKL is essential for utilizing information from multiple information sources and DE is essential for formulating a trading rule based on a mixture of discrete structures and continuous parameters. Initially, the prediction model optimized by MKL predicts the returns based on a technical indicator called the moving average convergence and divergence. Next, a combined trading signal is optimized by DE using the inputs from the prediction model and technical indicator RSI obtained from multiple timeframes. The experimental results showed that trading using the prediction learned by MKL yielded consistent profits.


Oral Microbiota Shift after 12-Week Supplementation with Lactobacillus reuteri DSM 17938 and PTA 5289; A Randomized Control Trial.
Lactobacillus spp. potentially contribute to health by modulating bacterial biofilm formation, but their effects on the overall oral microbiota remain unclear. Oral microbiota was characterized via 454-pyrosequencing of the 16S rDNA hypervariable region V3-V4 after 12 weeks of daily Lactobacillus reuteri DSM 17938 and PTA 5289 consumption. Forty-four adults were assigned to a test group (n = 22) that received lactobacilli lozenges (108 CFU of each strain/lozenge) or a control group that received placebo (n = 22). Presence of L. reuteri was confirmed by cultivation and species specific PCR. Tooth biofilm samples from 16 adults before, during, and after exposure were analyzed by pyrosequencing. A total of 1,310,292 sequences were quality filtered. After removing single reads, 257 species or phylotypes were identified at 98.5% identity in the Human Oral Microbiome Database. Firmicutes, Bacteroidetes, Fusobacteria, Proteobacteria, and Actinobacteria were the most abundant phyla. Streptococcus was the most common genus and the S. oralis/S. mitis/S. mitis bv2/S. infantis group comprised the dominant species. The number of observed species was unaffected by L. reuteri exposure. However, subjects who had consumed L. reuteri were clustered in a principal coordinates analysis relative to scattering at baseline, and multivariate modeling of pyrosequencing microbiota, and culture and PCR detected L. reuteri separated baseline from 12-week samples in test subjects. L. reuteri intake correlated with increased S. oralis/S. mitis/S. mitis bv2/S. infantis group and Campylobacter concisus, Granulicatella adiacens, Bergeyella sp. HOT322, Neisseria subflava, and SR1 [G-1] sp. HOT874 detection and reduced S. mutans, S. anginosus, N. mucosa, Fusobacterium periodicum, F. nucleatum ss vincentii, and Prevotella maculosa detection. This effect had disappeared 1 month after exposure was terminated. L. reuteri consumption did not affect species richness but induced a shift in the oral microbiota composition. The biological relevance of this remains to be elucidated. ClinicalTrials.gov NCT02311218.


Assessment of recovery in patients undergoing intravenous conscious sedation using bispectral analysis.
The Bispectral Index (BIS) has been recently shown to objectively predict the level of sedation in patients undergoing conscious sedation. It was the goal of this study to directly compare the recovery profile of patients where the BIS was used to monitor sedation with a control group where the monitor was not used. Forty patients undergoing third molar extractions under intravenous conscious sedation were randomly assigned to 2 groups. In both groups, induction of sedation was performed using a standard dose of fentanyl (1.5 microg/kg) and midazolam (0.05 mg/kg). Propofol was then given in 10 to 20 mg boluses until a clinically desirable sedation level was achieved. In 1 group, the BIS was then monitored continually during surgery using a microcomputer (Aspect-1050 Monitor, Aspect Co, Natick, MA) and recorded at 5-minute intervals. The anesthetist (N.A.S.) provided additional propofol boluses to maintain a BIS level of 70 to 80. In the other group, the BIS sensor was applied, but the monitor was not used. In this group, the sedation was modified, and additional propofol was given based solely on the anesthetist's subjective assessment of the desired level of sedation (Observer's Assessment of Alertness/Sedation [OAA/S] scale level 2 to 3). Additional boluses of 1 mg of midazolam were given during the procedure if patients required repeated boluses of propofol at less than 5-minute intervals to maintain the desired sedation level (BIS level of 70 to 80 or OAA/S level of 2 to 3). These additional midazolam boluses, as well as the time of the last sedative dose (propofol or midazolam) were recorded to study the effect of these factors on recovery. Of the 40 patients initially included in the study, 1 subject in the BIS-monitored group was excluded due to the loss of intravenous access at initiation of the case. For the remaining 39 subjects, 19 were assessed objectively using the BIS monitor, whereas 20 were assessed subjectively using the OAA/S scale. The BIS cases were slightly longer in duration than the OAA/S cases, lasting an average of 26 minutes versus 22 minutes. This difference was statistically nonsignificant (P =.19). Less propofol was used in the BIS cases, with an average of 98 mg for BIS cases versus 106 mg for OAA/S cases (P =.59). The total dose in mg/kg/min was significantly less in the BIS group (0.054 mg/kg/min) than in the OAA/S group (0.074 mg/kg/min; P =.0082). There was no significant difference in the amount of midazolam administered after induction between the 2 groups (P =.60). The surgeon, who was blinded to whether the monitor was used, ranked the third molar extractions more difficult in the BIS group (P =.05). However, patients in the BIS group were on average more cooperative, with better maintenance of muscle tone. The difference in these parameters were nonsignificant (P =.15 and .092, respectively). A positive Romberg test was obtained earlier in BIS patients, although this difference was nonsignificant (P =.097). The straight-line test was completed significantly sooner in BIS patients (P =.013). There was no significant difference between the BIS and OAA/S groups in perceptual speed (P =.55) or computation (P =.32). There was essentially no difference between groups in patient-assessed comfort or recall of the procedure. There were also no notable differences in anesthesia complications, return to activities of daily life, or pain medication use between the 2 groups. The BIS provides additional information for standard monitoring techniques that helps guide the administration of sedative-hypnotic agents. It appears that use of the BIS monitor can help to titrate the level of sedation so that less drugs are used to maintain the desired level. The trend toward an earlier return of motor function in BIS-monitored patients warrants further investigation.


Efficacy of moxalactam in an animal model of subcutaneous abscesses: penetration into infected sites and in vivo activity against Bacteroides fragilis.
Moxalactam was compared with alternative antimicrobial agents for in vivo activity against Bacteroides fragilis. Mice were challenged with one of 15 strains and evaluated by quantitative bacteriologic analysis of abscess contents after five days of treatment. Optimal results were achieved with clindamycin, moxalactam, and cefoxitin. The mean decrease in bacteria with these three drugs was significantly greater than with chloramphenicol, cefotaxime, carbenicillin, cephalothin, ampicillin, and cefoperazone. The penetration of antimicrobial agents into subcutaneous abscesses was examined. The mean peak level in sterile abscesses of moxalactam was 27% of the mean peak serum level, a result indicating relatively good penetration compared with other beta-lactam antimicrobial agents. Stability of moxalactam to the beta-lactamase of B. fragilis was demonstrated in vivo by comparing levels of biologically active drug in sterile and infected abscesses and in vitro by comparing levels of antibiotic incubated in sterile exudate and exudate infected with B. fragilis. Moxalactam compares favorably with clindamycin and cefoxitin in activity against B. fragilis.


Subtalar distraction bone block arthrodesis.
This retrospective study analyses the results of subtalar bone block distraction arthrodesis used in the treatment of late complications of calcaneal fractures, acute severely comminuted fractures, nonunion (and malunion) of attempted subtalar arthrodeses, avascular necrosis of the talus, and club-foot deformity. Of 39 patients (41 feet) who had this procedure, 35 (37 feet) returned for follow-up after a mean of 70 months (26 to 140). There were 24 men (25 feet) and 11 women (12 feet) with a mean age of 41 years (16 to 63). Each completed a standardised questionnaire, based on the hindfoot-scoring system of the American Orthopaedic Foot and Ankle Society and were reviewed both clinically and radiologically. Of the 37 operations, 32 (87%) achieved union. The mean hindfoot score (maximum of 94 points) increased from 21.1 points (8 to 46) preoperatively to 68.9 (14 to 82) at the final follow-up. The mean talocalcaneal and calcaneal pitch angles were 20.5 degrees and 4.9 degrees before operation, 25.9 degrees and 8.3 degrees immediately after, and 24.6 degrees and 7.7 degrees at the final follow-up, respectively. The mean talar declination angle improved from 6.5 degrees (-10 to 22) before operation to 24.8 degrees (14 to 32) at the final follow-up. The mean talocalcaneal height increased from 68.7 mm before operation to 74.5 mm immediately after and 73.5 mm at the final follow-up. Of the 37 arthrodeses available for review, 32 were successful; 29 patients (30 arthrodeses) were satisfied with the procedure. Minimal loss of hindfoot alignment occurred when comparing radiographs taken immediately after operation and at final follow-up.


Adaptive imaging on a diagnostic ultrasound scanner at quasi real-time rates.
Constructing an ultrasonic imaging system capable of compensating for phase errors in real-time is a significant challenge in adaptive imaging. We present a versatile adaptive imaging system capable of updating arrival time profiles at frame rates of approximately 2 frames per second (fps) with 1-D arrays and up to 0.81 fps for 1.75-D arrays, depending on the desired near-field phase correction algorithm. A novel feature included in this system is the ability to update the aberration profile at multiple beam locations for 1-D arrays. The features of this real-time adaptive imaging system are illustrated in tissue-mimicking phantoms with physical near-field phase screens and evaluated in clinical breast tissue with a 1.75-D array. The contrast-to-noise ratio (CNR) of anechoic cysts was shown to improve dramatically in the tissue-mimicking phantoms. In breast tissue, the width of point-like targets showed significant improvement: a reduction of 26.2% on average. Brightness of these targets, however, marginally decreased by 3.9%. For larger structures such as cysts, little improvement in features and CNR were observed, which is likely a result of the system assuming an infinite isoplanatic patch size for the 1.75-D arrays. The necessary requirements for constructing a real-time adaptive imaging system are also discussed.


Effect of Alirocumab on Lipoprotein(a) and Cardiovascular Risk After Acute Coronary Syndrome.
Lipoprotein(a) concentration is associated with cardiovascular events. Alirocumab, a proprotein convertase subtilisin/kexin type 9 inhibitor, lowers lipoprotein(a) and low-density lipoprotein cholesterol (LDL-C). A pre-specified analysis of the placebo-controlled ODYSSEY Outcomes trial in patients with recent acute coronary syndrome (ACS) determined whether alirocumab-induced changes in lipoprotein(a) and LDL-C independently predicted major adverse cardiovascular events (MACE). One to 12 months after ACS, 18,924 patients on high-intensity statin therapy were randomized to alirocumab or placebo and followed for 2.8 years (median). Lipoprotein(a) was measured at randomization and 4 and 12 months thereafter. The primary MACE outcome was coronary heart disease death, nonfatal myocardial infarction, ischemic stroke, or hospitalization for unstable angina. Baseline lipoprotein(a) levels (median: 21.2 mg/dl; interquartile range [IQR]: 6.7 to 59.6 mg/dl) and LDL-C [corrected for cholesterol content in lipoprotein(a)] predicted MACE. Alirocumab reduced lipoprotein(a) by 5.0 mg/dl (IQR: 0 to 13.5 mg/dl), corrected LDL-C by 51.1 mg/dl (IQR: 33.7 to 67.2 mg/dl), and reduced the risk of MACE (hazard ratio [HR]: 0.85; 95% confidence interval [CI]: 0.78 to 0.93). Alirocumab-induced reductions of lipoprotein(a) and corrected LDL-C independently predicted lower risk of MACE, after adjustment for baseline concentrations of both lipoproteins and demographic and clinical characteristics. A 1-mg/dl reduction in lipoprotein(a) with alirocumab was associated with a HR of 0.994 (95% CI: 0.990 to 0.999; p = 0.0081). Baseline lipoprotein(a) and corrected LDL-C levels and their reductions by alirocumab predicted the risk of MACE after recent ACS. Lipoprotein(a) lowering by alirocumab is an independent contributor to MACE reduction, which suggests that lipoprotein(a) should be an independent treatment target after ACS. (ODYSSEY Outcomes: Evaluation of Cardiovascular Outcomes After an Acute Coronary Syndrome During Treatment With Alirocumab; NCT01663402).


Use of Virtual Reality Tools for Vestibular Disorders Rehabilitation: A Comprehensive Analysis.
Classical peripheral vestibular disorders rehabilitation is a long and costly process. While virtual reality settings have been repeatedly suggested to represent possible tools to help the rehabilitation process, no systematic study had been conducted so far. We systematically reviewed the current literature to analyze the published protocols documenting the use of virtual reality settings for peripheral vestibular disorders rehabilitation. There is an important diversity of settings and protocols involving virtual reality settings for the treatment of this pathology. Evaluation of the symptoms is often not standardized. However, our results unveil a clear effect of virtual reality settings-based rehabilitation of the patients' symptoms, assessed by objectives tools such as the DHI (mean decrease of 27 points), changing symptoms handicap perception from moderate to mild impact on life. Furthermore, we detected a relationship between the duration of the exposure to virtual reality environments and the magnitude of the therapeutic effects, suggesting that virtual reality treatments should last at least 150 minutes of cumulated exposure to ensure positive outcomes. Virtual reality offers a pleasant and safe environment for the patient. Future studies should standardize evaluation tools, document putative side effects further, compare virtual reality to conventional physical therapy, and evaluate economical costs/benefits of such strategies.


Chemical Entities of Biological Interest: an update.
Chemical Entities of Biological Interest (ChEBI) is a freely available dictionary of molecular entities focused on 'small' chemical compounds. The molecular entities in question are either natural products or synthetic products used to intervene in the processes of living organisms. Genome-encoded macromolecules (nucleic acids, proteins and peptides derived from proteins by cleavage) are not as a rule included in ChEBI. In addition to molecular entities, ChEBI contains groups (parts of molecular entities) and classes of entities. ChEBI includes an ontological classification, whereby the relationships between molecular entities or classes of entities and their parents and/or children are specified. ChEBI is available online at http://www.ebi.ac.uk/chebi/. This article reports on new features in ChEBI since the last NAR report in 2007, including substructure and similarity searching, a submission tool for authoring of ChEBI datasets by the community and a 30-fold increase in the number of chemical structures stored in ChEBI.


Accumulation of mutations and somatic selection in aging neural stem/progenitor cells.
Genomic instability within somatic stem cells may lead to the accumulation of mutations and contribute to cancer or other age-related phenotypes. However, determining the frequency of mutations that differ among individual stem cells is difficult from whole tissue samples because each event is diluted in the total population of both stem cells and differentiated tissue. Here the ability to expand neural stem/progenitor cells clonally permitted measurement of genomic alterations derived from a single initial cell. C57Bl/6 x DBA/2 hybrid mice were used and PCR analysis with strain-specific primers was performed to detect loss of heterozygosity on nine different chromosomes for each neurosphere. The frequency with which changes occurred in neurospheres derived from 2-month- and 2-year-old mice was compared. In 15 neurospheres derived from young animals both parental chromosomes were present for all nine chromosome pairs. In contrast, 16/17 neurospheres from old animals demonstrated loss of heterozygosity (LOH) on one or more chromosomes and seven exhibited a complete deletion of at least one chromosomal region. For chromosomes 9 and 19 there is a significant bias in the allele that is lost where in each case the C57Bl/6 allele is retained in 6/6 neurospheres exhibiting LOH. These data suggest that aging leads to a substantial mutational load within the neural stem cell compartment which can be expected to affect the normal function of these cells. Furthermore, the retention of specific alleles for chromosomes 9 and 19 suggests that a subset of mutational events lead to an allele-specific survival advantage within the neural stem cell compartment.


Sampling expansions for three-dimensional light amplitude distribution in the vicinity of an axial image point: comment.
Landgrave and Berriel-Valdos presented axial and radial sampling expansions for three-dimensional light amplitude distribution around the Gaussian focal point. [J. Opt. Soc. Am. A 14, 2962 (1997)]. The expansions were obtained under the assumption that the pupil function was rotationally symmetric. We present a new derivation of the axial expansion that does not make use of arbitrary formal assumptions used by Landgrave and Berriel-Valdos and eliminates some faults of the derivation given by Arsenault and Boivin, who published this expansion in 1967 [J. Appl. Phys. 38, 3988 (1967)]. We also discuss generalizations of the axial expansion to the case of pupils that exhibit no symmetry with respect to the axis considered.


Risk factors of cellulitis in cirrhosis and antibiotic prophylaxis in preventing recurrence.
Cellulitis is a commonly encountered bacterial infection among cirrhotic patients apart from spontaneous bacterial peritonitis, urinary tract and respiratory infections. This study aimed to determine the risk factors of cellulitis in cirrhosis and whether antibiotic prophylaxis helps prevent recurrence of cellulitis. The study was conducted in two phases. In phase 1, all cirrhotic patients admitted with cellulitis from August 2011 to August 2013 were taken as cases (n=70) and cirrhotic patients without cellulitis were included as controls (n=73). Baseline demographic data, comorbidities and investigations were noted and compared. In phase 2, the cases of phase 1 were divided into two groups, based on initiation of antibiotic prophylaxis at the time of discharge and were followed up for six months for recurrence of cellulitis. The main etiology of cirrhosis was alcohol and 68% of cases were Child C. Factors which showed significance in univariate analysis were presence of diabetes mellitus, hepatic encephalopathy, platelet count, albumin level and model for end-stage liver disease (MELD) score. Using logistic regression, hepatic encephalopathy (OR 2.95, CI 1.01-8.45), albumin level <2.5 g/dL (OR 2.80, CI 1.32-5.92) and MELD >15 (OR 2.95, CI 1.39-6.27) emerged as significant factors associated with cellulitis. Cellulitis recurred in 20% and recurrence was significantly low among antibiotic prophylaxis group (15% vs. 50% P=0.048). Hypoalbuminemia, and high MELD score are the risk factors for cellulitis in cirrhosis. Antibiotic prophylaxis can reduce the recurrence of cellulitis as in the case of spontaneous bacterial peritonitis.


An eHealth program versus a standard care supervised health program and associated health outcomes in individuals with mobility disability: study protocol for a randomized controlled trial.
Young adults with mobility disability (MD) are less likely to engage in regular physical activity (PA) compared with their able-bodied peers and inactive adults with a MD are more likely to report one or more chronic diseases compared to those who are physically active. Despite the vast amount of research published in the field of PA interventions over the past decades, little attention has been focused on interventions aiming to increase PA among individuals with MD. Thus, we propose to compare the effects of an eHealth program compared to a usual care supervised health program on levels of PA and other health behaviors. The current intervention will use a randomized controlled trial (RCT) design with two treatment groups (an eHealth program and a usual care supervised health program) in young adults with newly acquired MD. In total, 110 young adults (aged 18-40 years) with a MD, acquired within the past 3 years, will be recruited to participate in a 12-week intervention. The primary study outcome is accelerometer-measured time spent in moderate to vigorous PA. Secondary outcomes includes health-related quality of life, depression, stress, fitness, body composition, diet, musculoskeletal pain, motivation to exercise and work ability. There is a lack of RCTs investigating effective ways to increase levels of PA in young adults with MD. Increased levels of PA among this physically inactive population have the potential to substantially improve health-related outcomes, possibly more so than in the general population. The trial will put strong emphasis on optimizing exercise adherence and investigating feasibility in the two treatment programs. The Ethical Review Board (EPN) at Karolinska Institutet has approved the study (2017/1206-31/1). International Standard Randomised Controlled Trial Number (ISRCTN), reference number ISRCTN22387524 . Prospectively registered February 4, 2018.


Risk factors for and outcomes of intraprocedural rupture during endovascular treatment of unruptured intracranial aneurysms.
The risk factors for intraprocedural rupture (IPR) of unruptured intracranial aneurysms (UIAs) and the outcomes of IPR itself are unclear. This study was performed to identify the independent risk factors for and outcomes of IPR. We retrospectively evaluated the medical records and radiologic data of 1375 patients (1406 UIAs) who underwent coil embolization from January 2001 to October 2016. IPR occurred in 20 aneurysms of 20 patients (1.4%). Univariate analyses showed that the rate of IPR was significantly higher in the treatment of aneurysms with a small dome size, aneurysms in the anterior communicating artery (AcomA) (6.6%), and patients with a medical history of dyslipidemia. Multivariate analyses showed that a small dome size and aneurysms in the AcomA were independently associated with IPR (p=0.0096 and p=0.0001, respectively). IPR induced by a microcatheter was associated with a higher risk of severe subarachnoid hemorrhage than other causes of IPR (57% vs 0%, respectively). Thromboembolic complications occurred in seven (35%) patients with IPR. Six (30%) patients required external ventricular drainage placement after developing symptoms of acute hydrocephalus. The overall morbidity and mortality rates from IPR were 0.22% and 0.15%, respectively. Aneurysms in the AcomA and with a small dome size are likely to be risk factors for IPR. IPR induced by microcatheters can result in poor outcomes. The rate of IPR-associated thromboembolic complications is high, and IPR itself is associated with acute hydrocephalus. If managed appropriately, however, most patients with IPR can survive without neurological deterioration.


Blood pressure and heart rate variability during yoga-based alternate nostril breathing practice and breath awareness.
Previous research has shown a reduction in blood pressure (BP) immediately after the practice of alternate nostril yoga breathing (ANYB) in normal healthy male volunteers and in hypertensive patients of both sexes. The BP during ANYB has not been recorded. Participants were 26 male volunteers (group mean age ±SD, 23.8±3.5 years). We assessed (1) heart rate variability, (2) non-invasive arterial BP, and (3) respiration rate, during (a) ANYB and (b) breath awareness (BAW) sessions. Each session was 25 minutes. We performed assessments at 3 time points: Pre (5 minutes), during (15 minutes; for ANYB or BAW) and Post (5 minutes). A naïve-to-yoga control group (n=15 males, mean age ±SD 26.1±4.0 years) were assessed while seated quietly for 25 minutes. During ANYB there was a significant decrease (repeated measures ANOVA) in systolic BP and respiration rate; while RMSSD (the square root of the mean of the sum of squares of differences between adjacent NN intervals) and NN50 (the number of interval differences of successive normal to normal intervals greater than 50 ms) significantly increased. During BAW respiration rate decreased. In contrast, respiration rate increased during the control state. ANYB and BAW were significantly different (2-factor ANOVA) in RMSSD and respiration rate. BAW and control were different with respect to respiration rate. The results suggest that vagal activity increased during and after ANYB, which could have contributed to the decrease in BP and changes in the HRV.


Iridium nanoparticles stabilized by metal organic frameworks (IrNPs@ZIF-8): synthesis, structural properties and catalytic performance.
Today, the synthesis of well-defined metal nanoparticles stabilized by the metal-organic frameworks (MOFs), which provide high specific surface areas, tunable pore sizes, and guest interactable organic linkers, and the discovery of their unique properties are still challenging goals. The chemically robust zeolitic imidazole framework (ZIF) is a subclass of MOF. In this study, the microporous sodalite-like ZIF-8 (Zn(MeIM); MeIM = 2-methylimidazole) was selected as host matrix to stabilize guest iridium nanoparticles (IrNPs). The iridium loading was achieved via gas phase infiltration of Ir(COD)(MeCp) (methylcyclopentadienyl)(1,5-cyclooctadiene)iridium(i)) precursor followed by hydrogenolysis of the inclusion compound Ir(COD)(MeCp)@ZIF-8 to form the IrNPs@ZIF-8. The characterization of IrNPs@ZIF-8 by ICP-MS, P-XRD, XPS, (13)C MAS NMR, TEM, HRTEM, STEM, STEM-EDX, HAADF-STEM, DR-UV-vis, EA analyses and N(2)-adsorption-desorption techniques reveal the formation of well-dispersed iridium nanoparticles (3.3 ± 1.7 nm) within the framework of ZIF-8 (IrNPs@ZIF-8) by keeping the host framework intact. The catalytic application of IrNPs@ZIF-8 in terms of activity, selectivity, reusability and durability was demonstrated in the hydrogenation of cyclohexene and phenylacetylene under mild conditions, in which they were found to be highly active catalysts. Moreover, they show great durability against sintering and leaching throughout the catalytic runs that make them highly reusable catalysts. They retain their inherent catalytic activity even at the fifth catalytic run in the hydrogenation of cyclohexene and phenylacetylene.


Predictors of carotid atherosclerosis in systemic lupus erythematosus.
Patients with systemic lupus erythematosus (SLE) are at increased risk for cardiovascular and cerebrovascular events, even after adjustment for traditional risk factors. We examined the association of traditional risk factors, novel markers of cardiovascular disease (C-reactive protein, homocysteine, lipoprotein(a), plasminogen activator inhibitor-1, fibrinogen), and markers indicative of SLE activity (including C3, C4, anti-dsDNA, and prednisone use) with the presence of significant plaque on carotid duplex imaging. Six hundred five patients with SLE enrolled in the Hopkins Lupus Cohort Study (92% female, 38% African-American) underwent carotid duplex testing. Prospectively gathered clinical, laboratory, and serologic data from their quarterly followup visits in the Hopkins Lupus Cohort were used in the analyses. For predictors that varied over time, such as cholesterol, the mean values during cohort participation were calculated for the analysis. Informed consent was obtained from all patients. The presence of carotid plaque was strongly associated with age, ranging from 1% among those less than 30 years of age to 61% among those 60 years or older. After adjusting for age, there were moderate or strong associations of carotid plaque with male gender (age-adjusted risk 25% vs 13%; p = 0.051), hypertension (age-adjusted risk 18% vs 8%; p = 0.0001), diabetes mellitus (age-adjusted risk 19% vs 13%; p = 0.075), C3 > 120 mg/dl (age-adjusted risk 18% vs 11% and 14% for normal and low C3, respectively; p = 0.046), serum creatinine > 1.3 (age-adjusted risk 32% vs 13%; p = 0.039), and mean systolic blood pressure > 140 (age-adjusted risk 23% vs 13%; p = 0.028). There was no strong evidence of an association between plaque and SLE disease activity (age-adjusted risk 14% among those with adjusted mean SLEDAI > 3 vs 14% among those with lower SLEDAI) or with time since SLE diagnosis (age-adjusted risk 12%, 14%, and 16% among those with SLE for < 2, 2-8, and > 8 years, respectively; p = 0.49). Traditional cardiovascular risk factors were associated with carotid plaque in SLE. However, SLE disease activity and duration of SLE are not strongly associated with carotid plaque. A "lupus factor" separate from traditional risk factors remains unidentified.


Internet-based cognitive behavior therapy for obsessive compulsive disorder: a pilot study.
Cognitive behavior therapy (CBT) is widely regarded as an effective treatment for obsessive compulsive disorder (OCD), but access to CBT therapists is limited. Internet-based CBT (ICBT) with therapist support is a way to increase access to CBT but has not been developed or tested for OCD. The aim of this study was to evaluate ICBT for OCD. An open trial where patients (N = 23) received a 15-week ICBT program with therapist support consisting of psychoeducation, cognitive restructuring and exposure with response prevention. The primary outcome was the Yale-Brown Obsessive Compulsive Scale (Y-BOCS), which was assessed by a psychiatrist before and immediately after treatment. Secondary outcomes were self-rated measures of OCD symptoms, depressive symptoms, general functioning, anxiety and quality of life. All assessments were made at baseline and post-treatment. All participants completed the primary outcome measure at all assessment points. There were reductions in OCD symptoms with a large within-group effect size (Cohen's d = 1.56). At post-treatment, 61% of participants had a clinically significant improvement and 43% no longer fulfilled the diagnostic criteria of OCD. The treatment also resulted in statistically significant improvements in self-rated OCD symptoms, general functioning and depression. ICBT with therapist support reduces OCD symptoms, depressive symptoms and improves general functioning. Randomized trials are needed to confirm the effectiveness of this new treatment format.


Charting a path for innovative toilet technology using multicriteria decision analysis.
Practical and theoretically sound methods for analyzing innovative environmental technologies are needed to inform public and private decisions regarding research and development, risk management, and stakeholder communication. By integrating scientific assessments with a characterization of values, multicriteria decision analysis (MCDA) supports the ranking of alternative technology pathways on the basis of technical, financial, and social concerns. We applied MCDAto evaluate the use of NoMix urine separating toilets for managing environmental risk and postponing expensive upgrades to a large wastewater treatment plant near Zürich, Switzerland. Results indicate that, given current priorities, no single, fixed course of action (including the status quo) will be desirable to all stakeholders over the considered time horizon. However, a path forward is suggested that is not significantly disadvantageous to any stakeholder now and leaves open future options, allowing society to achieve overall greater benefits if priorities change, new environmental risks are revealed, or technology improves. While our analysis focuses on a particular catchment in Switzerland, many communities worldwide are faced with an aging and inefficient wastewater treatment infrastructure while also experiencing growth and development. Our framework can help these communities balance the conflicting objectives of diverse stakeholders and gain insight into the role that urine separation can play in transitioning to a more comprehensive and sustainable urban water management system.


Management of acute coronary syndromes. Variations in practice and outcome; findings from the Global Registry of Acute Coronary Events (GRACE).
Despite advances in the treatment of acute coronary syndromes based on randomized trial data and published guidelines, the extent to which such treatments are applied in practice remains uncertain. Data from clinical trials derive from selected geographical areas and in highly selected populations of patients, and hence may not reflect the overall population. The aim of the study was to investigate variations in hospital management and outcome using unselected data collected in the prospective Global Registry of Acute Coronary Events (GRACE). The 95 hospitals in GRACE were organized into 18 population-based clusters in 14 countries. Information was recorded about patient management and outcome during hospitalization and after discharge. Data on treatments administered were analysed by baseline condition, hospital type, by the presence or absence of a catheterization laboratory, and by geographical region. Of 11543 patients, 44% had an admission diagnosis of unstable angina, 36% presented with myocardial infarction, 9% were admitted to rule out a myocardial infarction, 7% had chest pain and 4% were hospitalized for 'other cardiac' and 'non-cardiac' diagnoses. Of the total GRACE population 38% had a final diagnosis of unstable angina, 30% ST-segment elevation myocardial infarction, 25% non-ST-segment elevation myocardial infarction, and 7% of 'other cardiac' and 'non-cardiac' final diagnoses. The event rates for hospital death or reinfarction were six and 2% for non-ST-segment elevation myocardial infarction, seven and 3% for ST-segment elevation myocardial infarction, and 3% hospital death for unstable angina. The use of aspirin was similar across all hospital types and geographical regions. In contrast, the use of percutaneous coronary intervention and glycoprotein IIb/IIIa inhibitors was higher (P<0.0001) in teaching hospitals and hospitals with catheterization laboratories and was also higher in the United States. At discharge a higher percentage (P<0.0001) of patients received angiotensin-converting enzyme inhibitors in hospitals without catheterization laboratories. The use of statins was lower in non-teaching hospitals and in centres without a catheterization laboratory. The GRACE study reveals substantial differences in the management of patients based on hospital type and geographical location. Further analyses will determine whether such variations translate into differences in longer term outcomes. GRACE provides a multinational reference for the implementation of therapies of proven efficacy.


Pluralistic ignorance and alcohol use on campus: some consequences of misperceiving the social norm.
Four studies examined the relation between college students' own attitudes toward alcohol use and their estimates of the attitudes of their peers. All studies found widespread evidence of pluralistic ignorance: Students believed that they were more uncomfortable with campus alcohol practices than was the average student. Study 2 demonstrated this perceived self-other difference also with respect to one's friends. Study 3 traced attitudes toward drinking over the course of a semester and found gender differences in response to perceived deviance: Male students shifted their attitudes over time in the direction of what they mistakenly believed to be the norm, whereas female students showed no such attitude change. Study 4 found that students' perceived deviance correlated with various measures of campus alienation, even though that deviance was illusory. The implications of these results for general issues of norm estimation and responses to perceived deviance are discussed.


Porcine kidney and heart transplantation in baboons undergoing a tolerance induction regimen and antibody adsorption.
Xenotransplantation would provide a solution to the current shortage of organs for transplantation. Our group has been successful in inducing tolerance in mice and monkey models of allogeneic transplantation. The present study attempts to extend the same tolerance-inducing regimen to a pig-to-baboon organ transplantation model. Nine baboons underwent a conditioning regimen (consisting of nonmyeloablative or myeloablative whole body and thymic irradiation, splenectomy, antithymocyte globulin, pharmacologic immunosuppression and porcine bone marrow transplantation [BMTx]), which has previously been demonstrated to induce donor-specific allograft tolerance in monkeys. In addition, immunoadsorption of anti-alphaGal antibody (Ab) was performed. Four of the nine baboons received pig kidney transplants (KTx), and one also underwent repeat transplantation with an SLA-matched kidney. Two received heterotopic pig heart transplants (HTx). Three baboons underwent conditioning without organ transplantation for long-term studies of natural Ab kinetics. In the three baboons that received the conditioning regimen without an organ transplant, immunoadsorption reduced Ab by approximately 90%, but recovery of Ab to pretreatment level or higher occurred within 7 days. In contrast, the level of Ab remained low after organ transplant. No Ab to pig antigens other than alphaGal was detected in any baboon before or after BMTx, KTx, or HTx. No graft succumbed to hyperacute rejection. KTx function began to deteriorate within 3-6 days, with oliguria and hematuria progressing to anuria, and the kidneys were excised after 3, 6, 9, 11, and 14 days, respectively. One HTx ceased functioning at 8 days; the second baboon died with a contracting HTx at 15 days. Features of coagulopathy and thrombocytopenia developed in all six transplanted baboons (high D-dimer, prolonged prothrombin time and partial thromboplastin time, and falling fibrinogen) resulting in serious bleeding complications in two baboons, one of which died on day 9. Donor organs showed progressive acute humoral rejection with deposits of IgM, IgG, and complement; a focal mononuclear cellular infiltrate was also observed. The ureter was the earliest structure of the KTx affected by rejection, with progression to necrosis. This conditioning regimen prevented hyperacute rejection but was ineffective in preventing the return of Ab, which was associated with the development of acute humoral rejection with features of coagulopathy. No baboon developed anti-pig Ab other than alphaGal Ab. Further modifications of the protocol directed toward suppression of production of Ab are required to successfully induce tolerance to pig organs in baboons.


Enzyme-nanoporous gold biocomposite: excellent biocatalyst with improved biocatalytic performance and stability.
Applications involving biomolecules, such as enzymes, antibodies, and other proteins as well as whole cells, are often hampered by their unstable nature at extremely high temperature and in organic solvents. We constructed enzyme-NPG biocomposites by assembling various enzymes onto the surface of nanoporous gold (NPG), which showed much enhanced biocatalytic performance and stability. Various enzymes with different molecular sizes were successfully tethered onto NPG, and the loadings were 3.6, 3.1 and 0.8 mg g(-1) for lipase, catalase and horseradish peroxidase, respectively. The enzyme-NPG biocomposites exhibited remarkable catalytic activities which were fully comparable to those of free enzymes. They also presented enhanced stability, with 74, 78 and 53% of enzymatic activity retained after 20 successive batch reactions. Moreover, these novel biocomposites possessed significantly enhanced reaction durability under various thermal and in organic solvent systems. In a sample transesterification reaction, a high conversion rate was readily achieved by using the lipase-NPG biocomposite. These nano-biocomposite materials hold great potential in applications such as biosensing, molecular electronics, catalysis, and controlled delivery.


Are perceived muscle tension, electromyographic hyperactivity and personality traits correlated in the fibromyalgia syndrome?
The study was performed to investigate the relationship between perceived muscle tension and electromyographic hyperactivity and to what extent electromyographic (EMG) hyperactivity relates to personality traits in fibromyalgics. Thirty-six females with fibromyalgia performed isokinetic maximal forward flexions of the shoulder combined with surface EMG recordings of the trapezius and infraspinatus muscles. Signal amplitude ratio and peak torque were calculated in the initial and endurance test phases. Pain intensity, perceived general and local shoulder muscle tension, and personality traits using the Karolinska Scales of Personality were assessed pre-test. Neither perceived muscle tension nor muscular tension personality trait correlated with EMG muscle hyperactivity. Perceived general muscle tension correlated with aspects of anxiety proneness (including muscle tension) of the Karolinska Scales of Personality. Pain intensity interacted with many of the variables. We propose that when patients with fibromyalgia report muscle tension that they may be expressing something other than physiological muscle tension.


Complete hydatidiform mole coexisting with a live fetus.
The co-existence of a hydatidiform mole with a living fetus is a rare phenomenon. The condition is a dilemma with respect to the diagnosis and management of associated maternal (a risk of maternal complications, such as preeclampsia, hyperthyrodism, and a risk of malignancy) and fetal (elevated risk of spontaneous abortion, neonatal thyrotoxicosis) complications. A 27-year-old woman was referred to our hospital with a diagnosis of hydatidiform mole and live fetus. The pregnancy was unremarkable except for the complaints of excessive nausea and vomiting. Successive ultrasound examinations demonstrated a normally growing live fetus (14 weeks) alongside a normal placenta and an additional intrauterine echogenic mass with features of hydatidiform mole. Genetic amniocentesis at 18 weeks' of gestation showed normal diploid fetal karyotype. At 20 weeks of pregnancy, a control prenatal visit revealed intrauterine fetal death. The follow-up period for two years was unremarkable. In the case of a normal fetal karyotype and the absence of serious signs of maternal pathology, waiting until fetal viability is achieved can justifiably be proposed, however there is still a risk of prenatal complications such as intrauterine death.


Design of aqueous two-phase systems for purification of hyaluronic acid produced by metabolically engineered Lactococcus lactis.
Hyaluronic acid has a wide range of biomedical applications and its commercial value is highly dependent on its purity and molecular weight. This study highlights the utility of aqueous two-phase separation as a primary recovery step for hyaluronic acid and for removal of major protein impurities from fermentation broths. Metabolically engineered cultures of a lactate dehydrogenase mutant strain of Lactococcus lactis (L. lactis NZ9020) were used to produce high-molecular-weight hyaluronic acid. The cell-free fermentation broth was partially purified using a polyethylene glycol/potassium phosphate system, resulting in nearly 100% recovery of hyaluronic acid in the salt-rich bottom phase in all the aqueous two-phase separation experiments. These experiments were optimized for maximum removal of protein impurities in the polyethylene glycol rich top phase. The removal of protein impurities resulted in substantial reduction of membrane fouling in the subsequent diafiltration process, carried out with a 300 kDa polyether sulfone membrane. This step resulted in considerable purification of hyaluronic acid, without any loss in recovery and molecular weight. Diafiltration was followed by an adsorption step to remove minor impurities and achieve nearly 100% purity. The final hyaluronic acid product was characterized by Fourier-transform IR and NMR spectroscopy, confirming its purity.


Comparison of the abilities of Salmonella typhimurium rpoS, aroA and rpoS aroA strains to elicit humoral immune responses in BALB/c mice and to cause lethal infection in athymic BALB/c mice.
Salmonella typhimurium rpoS and rpoS aroA mutants are effective live vaccines in the murine model of salmonellosis (Coynault et al., Mol. Microbiol. 1996; 22: 149-60). Here, we further investigate the characteristics of these vaccines. The systemic humoral response induced by S. typhimurium rpoS, aroA and rpoS aroA vaccine candidates against S. typhimurium LPS was studied by ELISA. In BALB/c mice, the rpoS aroA strain induced a systemic anti-LPS humoral response similar to that induced by the rpoS and aroA strains. The virulence of aroA and rpoS aroA vaccines in nude (nu/nu) BALB/c mice was also compared. Salmonella typhimurium aroA and rpoS aroA vaccines both produced slowly progressing lethal infections in athymic mice inoculated i.p. but the rpoS aroA strain was more attenuated than the aroA strain, as determined by time to death and bacterial counts in spleens. Finally, a rpoS mutant of Salmonella dublin conferred protection in mice against an oral challenge with a wild-type strain of S. dublin whereas a rpoS mutant of S. typhimurium did not. This suggests that the protection provided by the S. typhimurium rpoS vaccine is serotype-dependent.


Receptor-mediated transcytosis of IgA in MDCK cells is via apical recycling endosomes.
Classically, the polymeric immunoglobulin receptor and its ligand, IgA, are thought to be sorted from basolateral early endosomes into transcytotic vesicles that directly fuse with the apical plasma membrane. In contrast, we have found that in MDCK cells IgA is delivered from basolateral endosomes to apical endosomes and only then to the apical cell surface. When internalized from the basolateral surface of MDCK cells IgA is found to accumulate under the apical plasma membrane in a compartment that is accessible to two apically added membrane markers: anti-secretory component Fab fragments, and avidin internalized from the biotinylated apical pole of the cell. This accumulation occurs in the presence of apical trypsin, which prevents internalization of the ligand from the apical cell surface. Using a modification of the diaminobenzidine density-shift assay, we estimate that approximately 80% of basolaterally internalized IgA resides in the apical endosomal compartment. In addition, approximately 50% of basolaterally internalized transferrin, a basolateral recycling protein, has access to this apical endosomal compartment and is efficiently recycled back to the basolateral surface. Microtubules are required for the organization of the apical endosomal compartment and it is dispersed in nocodazole-treated cells. Moreover, this compartment is largely inaccessible to fluid-phase markers added to either pole of the cell, and therefore seems analogous to the recycling endosome described in nonpolarized cells. We propose a model in which transcytosis is not a specialized pathway that uses unique transcytotic vesicles, but rather combines portions of pathways used by non-transcytosing molecules.


Effect of Flowering Time-Related Genes on Biomass, Harvest Index, and Grain Yield in CIMMYT Elite Spring Bread Wheat.
Grain yield (YLD) is a function of the total biomass (BM) and of partitioning the biomass by grains, i.e., the harvest index (HI). The most critical developmental stage for their determination is the flowering time, which mainly depends on the vernalization requirement (Vrn) and photoperiod sensitivity genes (Ppd) loci. Allelic variants at the Vrn, Ppd, and earliness per se (Eps) genes of elite spring wheat genotypes included in High Biomass Association Panel (HiBAP) I and II were used to estimate their effects on the phenological stages BM, HI, and YLD. Each panel was grown for two consecutive years in Northwest Mexico. Spring alleles at Vrn-1 had the largest effect on shortening the time to anthesis, and the Ppd-insensitive allele Ppd-D1a had the most significant positive effect on YLD in both panels. In addition, alleles at TaTOE-B1 and TaFT3-B1 promoted between 3.8% and 7.6% higher YLD and 4.2% and 10.2% higher HI in HiBAP I and II, respectively. When the possible effects of the TaTOE-B1 and TaFT3-B1 alleles on the sink and source traits were explored, the favorable allele at TaTOE-B1 showed positive effects on several sink traits mainly related to grain number. The favorable alleles at TaFT3-B1 followed a different pattern, with positive effects on the traits related to grain weight. The results of this study expanded the wheat breeders' toolbox in the quest to breed better-adapted and higher-yielding wheat cultivars.


A study in vitro of the sodium pump in fulminant hepatic failure.
1. The mechanism underlying the raised leucocyte sodium content in fulminant hepatic failure was studied by measurement of sodium fluxes, (Na+ + K+)-dependent adenosine triphosphatase activity, and leucocyte ATP content. 2. The rate constant for sodium efflux in the leucocytes was significantly reduced, and attributable to reduced activity of the enzyme (Na+ + K+)-ATPase. Leucocyte ATP content was not significantly different from that of control cells. 3. Incubation of cells from patients in the sera of normal subjects resulted in a reversal of these changes. Inhibition of the leucocyte sodium efflux rate constants and (Na+ +K+)-ATPase of normal cells was achieved by incubation in sera from patients. 4. We suggest that the raised sodium content of leucocytes in fulminant hepatic failure is attributable to a defective sodium pumping mechanism, possibly due to a circulating toxin.


Comparison of structure- and ligand-based scoring functions for deep generative models: a GPCR case study.
Deep generative models have shown the ability to devise both valid and novel chemistry, which could significantly accelerate the identification of bioactive compounds. Many current models, however, use molecular descriptors or ligand-based predictive methods to guide molecule generation towards a desirable property space. This restricts their application to relatively data-rich targets, neglecting those where little data is available to sufficiently train a predictor. Moreover, ligand-based approaches often bias molecule generation towards previously established chemical space, thereby limiting their ability to identify truly novel chemotypes. In this work, we assess the ability of using molecular docking via Glide-a structure-based approach-as a scoring function to guide the deep generative model REINVENT and compare model performance and behaviour to a ligand-based scoring function. Additionally, we modify the previously published MOSES benchmarking dataset to remove any induced bias towards non-protonatable groups. We also propose a new metric to measure dataset diversity, which is less confounded by the distribution of heavy atom count than the commonly used internal diversity metric. With respect to the main findings, we found that when optimizing the docking score against DRD2, the model improves predicted ligand affinity beyond that of known DRD2 active molecules. In addition, generated molecules occupy complementary chemical and physicochemical space compared to the ligand-based approach, and novel physicochemical space compared to known DRD2 active molecules. Furthermore, the structure-based approach learns to generate molecules that satisfy crucial residue interactions, which is information only available when taking protein structure into account. Overall, this work demonstrates the advantage of using molecular docking to guide de novo molecule generation over ligand-based predictors with respect to predicted affinity, novelty, and the ability to identify key interactions between ligand and protein target. Practically, this approach has applications in early hit generation campaigns to enrich a virtual library towards a particular target, and also in novelty-focused projects, where de novo molecule generation either has no prior ligand knowledge available or should not be biased by it.


Inhibition of prostaglandin synthesis in newborn pigs increases cerebral microvessel prostaglandin F2 alpha and prostaglandin E2 receptors, their second messengers and vasoconstrictor response to adult levels.
We recently reported that the density of prostaglandin (PG) F2 alpha and E2 receptors (FP and EP) on the cerebral microvasculature of the newborn is less than on that of the adult animal. This study tests the hypothesis that higher levels of PGF2 alpha and PGE2 in the newborn than in the adult brain might down-regulate FP and EP and their functions in the cerebral microvasculature. Newborn pigs (1-2 days old) were treated with ibuprofen (40 mg/kg i.v.) every 6 h for 48 h; and cerebrovascular FP and EP density, receptor-coupled second messenger production and cerebral vasoconstrictor responses to PGF2 alpha and PGE2 were determined. The results showed that ibuprofen treatment in the newborn increased brain microvascular FP and EP densities to levels found in the brains of adult pigs. This up-regulation of prostaglandin receptors was also observed in isolated newborn brain microvessels incubated for 48 h with ibuprofen. PGF 2 alpha, fenprostalene (PGF2 alpha analog), PGE2, 17-phenyl trinor PGE2 (EP1 receptor subtype agonist) and M&B 28,767 (EP3 agonist) caused a significantly greater increase in inositol 1,4,5-triphosphate production in brain microvessels of ibuprofen-treated than in brain microvessels of saline-treated newborn pigs. The cerebral vasoconstrictor effects of PGF2 alpha, 17-phenyl trinor PGE2 and M&B 28,767 were also significantly increased in newborn pigs treated with ibuprofen to levels comparable to those of adults. However, the steady-state level of FP mRNA in cerebral microvasculature did not differ between saline-treated newborn, ibuprofen-treated newborn and adult pigs. It is concluded that the low FP and EP densities in newborn brain microvessels are a result of high levels of brain prostaglandins and that these receptors, receptor-coupled second messengers and cerebral vasoconstrictor responses to FP, EP1 and EP3 stimulation can be up-regulated to adult levels by decreasing endogenous prostaglandin production. The changes in receptor levels were not related to steady-state levels of receptor mRNA in brain microvessels.


Suicide trends in discharged patients with mood disorders: associations with selective serotonin uptake inhibitors and comorbid substance misuse.
We determined the rates of suicide in two consecutive time intervals of low and high selective serotonin reuptake inhibitor (SSRI) prescribing for men and women with mood disorders, who were at relatively high risk of suicide, in a discharged hospitalized sample. High quality Swedish national registers were cross-linked to estimate rates of suicide. The results were stratified by three age-bands, and rates of comorbid substance misuse were explored. These were compared with general population suicide rates. There were 1606 suicides in discharged patients with mood disorders. Suicide rates decreased for all women during 1989-94 and 1995-2000 [odds ratio (OR) = 0.74, 95% confidence interval (CI) = 0.64-0.85] and a trend was found towards lower rates in men (OR = 0.89, 95% CI = 0.78-1.02). Approximately 90% of suicides occurred in those aged over 40 years; thus, the inpatient sample lacked the power to discriminate clear effects in the younger age groups. There was no increase or decrease in the rates of substance use disorders in patients who committed suicide. There were 20 851 suicides in the general population during 1989-2000. Suicide rates were lower at all ages for men and women in the general population in the interval of high SSRI prescribing. We conclude that there is no evidence from this study of an increase in suicide rates following the introduction of SSRIs in the general population or in a high-risk inpatient sample.


Establishing a Cancer Research Consortium in Low- and Middle-Income Countries: Challenges Faced and Lessons Learned.
There is an increasing effort in the global public health community to strengthen research capacity in low- and middle-income countries, but there is no consensus on how best to approach such endeavors. Successful consortia that perform research on HIV/AIDS and other infectious diseases exist, but few papers have been published detailing the challenges faced and lessons learned in setting up and running a successful research consortium. Members of the African Research Group for Oncology (ARGO) participated in generating lessons learned regarding the foundation and maintenance of a cancer research consortium in Nigeria. Drawing on our experience of founding ARGO, we describe steps and key factors needed to establish a successful collaborative consortium between researchers from both high- and low-income countries. In addition, we present challenges we encountered in building our consortium, and how we managed those challenges. Although our research group is focused primarily on cancer, many of our lessons learned can be applied more widely in biomedical or public health research in low-income countries. As the need for cancer care in LMICs continues to grow, the ability to create sustainable, innovative, collaborative research groups will become vital. Assessing the successes and failures that occur in creating and sustaining research consortia in LMICs is important for expansion of research and training capacity in LMICs.


Upstream tissue inhibitor of metalloproteinases-1 (TIMP-1) element-1, a novel and essential regulatory DNA motif in the human TIMP-1 gene promoter, directly interacts with a 30-kDa nuclear protein.
Elevated expression of the tissue inhibitor of metalloproteinases-1 (TIMP-1) protein and mRNA has been reported in human diseases including cancers and tissue fibrosis. Regulation of TIMP-1 gene expression is mainly mediated at the level of gene transcription and involves the activation of several well known transcription factors including those belonging to the AP-1, STAT, and Pea3/Ets families. In the current study, we have used DNase-1 footprinting to identify a new regulatory element (5'-TGTGGTTTCCG-3') present in the human TIMP-1 gene promoter. Mutagenesis and transfection studies in culture-activated rat hepatic stellate cells and the human Jurkat T cell line demonstrated that the new element named upstream TIMP-1 element-1 (UTE-1) is essential for transcriptional activity of the human TIMP-1 promoter. Electrophoretic mobility shift assay studies revealed that UTE-1 can form protein-DNA complexes of distinct mobilities with nuclear extracts from a variety of mammalian cell types and showed that induction of a high mobility UTE-1 complex is associated with culture activation of freshly isolated rat hepatic stellate cells. A combination of UV-cross-linking and Southwestern blotting techniques demonstrated that UTE-1 directly interacts with a 30-kDa nuclear protein that appears to be present in all cell types tested. We conclude that UTE-1 is a novel regulatory element that in combination with its cellular binding proteins may be an important component of the mechanisms controlling TIMP-1 expression in normal and pathological states.


Vaccinia virus replication is independent of cellular HSP72 expression which is induced during virus infection.
HSP72 is dramatically induced in the ovaries of vaccinia virus (VV)-infected mice and associates with VV proteins. In order to investigate the role of HSP72 during vaccinia virus replication, we have constructed a recombinant vaccinia virus encoding the major inducible cellular HSP72 (VV-HSP72+) and examined the replication characteristics of this virus. VV-HSP72+ exhibited growth kinetics identical to and peak titers very similar to those of control viruses, both in vitro and in vivo. In particular, replication of VV-HSP72+ was identical to that of control viruses in the HSP72-negative cell line Y3.Ag.1.2.3, and overexpression of HSP72 had no effect on the virulence of VV infection in normal or immunocompromised mice. We conclude that while VV infection results in the induction of the major inducible 72-kDa HSP, VV replication proceeds normally in the absence of this protein. It is unclear whether another celluar chaperone is required to facilitate virus replication in place of HSP72 in Y3.Ag.1.2.3 cells or whether HSP expression plays no role in virus replication, but is simply a component of the generalized stress response to virus infection.