Epichloë fungal endophyte interactions in perennial ryegrass (Lolium perenne L.) modified to accumulate foliar lipids for increased energy density.
Commercial cultivars of perennial ryegrass infected with selected Epichloë fungal endophytes are highly desirable in certain pastures as the resulting mutualistic association has the capacity to confer agronomic benefits (such as invertebrate pest deterrence) largely due to fungal produced secondary metabolites (e.g., alkaloids). In this study, we investigated T2 segregating populations derived from two independent transformation events expressing diacylglycerol acyltransferase (DGAT) and cysteine oleosin (CO) genes designed to increase foliar lipid and biomass accumulation. These populations were either infected with Epichloë festucae var. lolii strain AR1 or Epichloë sp. LpTG-3 strain AR37 to examine relationships between the introduced trait and the endophytic association. Here we report on experiments designed to investigate if expression of the DGAT + CO trait in foliar tissues of perennial ryegrass could negatively impact the grass-endophyte association and vice versa. Both endophyte and plant characters were measured under controlled environment and field conditions. Expected relative increases in total fatty acids of 17-58% accrued as a result of DGAT + CO expression with no significant difference between the endophyte-infected and non-infected progeny. Hyphal growth in association with DGAT + CO expression appeared normal when compared to control plants in a growth chamber. There was no significant difference in mycelial biomass for both strains AR1 and AR37, however, Epichloë-derived alkaloid concentrations were significantly lower on some occasions in the DGAT + CO plants compared to the corresponding null-segregant progenies, although these remained within the reported range for bioactivity. These results suggest that the mutualistic association formed between perennial ryegrass and selected Epichloë strains does not influence expression of the host DGAT + CO technology, but that endophyte performance may be reduced under some circumstances. Further investigation will now be required to determine the preferred genetic backgrounds for introgression of the DGAT + CO trait in combination with selected endophyte strains, as grass host genetics is a major determinant to the success of the grass-endophyte association in this species.


Synthesis of Novel Phosphorus-Containing Derivatives of 1,3,4-Trimethylglycoluril via the Birum-Oleksyszyn Reaction.
This work presents the synthesis of a new compound, 1-[aryl-(diphenylphosphono)methyl]-3,4,6-trimethylglycolurils, via the interaction of benzaldehyde and its mononitro- and monohydroxyderivatives with 1,3,4-trimethylglycoluril and triphenylphosphite. By varying the reaction conditions and the catalysts, the obtained product yields ranged from satisfactory to good. The diastereomers formed during the reaction were separated by semipreparative HPLC on the C18 stationary phase. The isolated diastereomers were characterized by 1H, 13C, and 31P NMR, and the structures of the diastereomers were confirmed using a single-crystal X-ray crystal structure analysis and quantum chemical calculations.


Whole heart coronary imaging with flexible acquisition window and trigger delay.
Coronary magnetic resonance imaging (MRI) requires a correctly timed trigger delay derived from a scout cine scan to synchronize k-space acquisition with the quiescent period of the cardiac cycle. However, heart rate changes between breath-held cine and free-breathing coronary imaging may result in inaccurate timing errors. Additionally, the determined trigger delay may not reflect the period of minimal motion for both left and right coronary arteries or different segments. In this work, we present a whole-heart coronary imaging approach that allows flexible selection of the trigger delay timings by performing k-space sampling over an enlarged acquisition window. Our approach addresses coronary motion in an interactive manner by allowing the operator to determine the temporal window with minimal cardiac motion for each artery region. An electrocardiogram-gated, k-space segmented 3D radial stack-of-stars sequence that employs a custom rotation angle is developed. An interactive reconstruction and visualization platform is then employed to determine the subset of the enlarged acquisition window for minimal coronary motion. Coronary MRI was acquired on eight healthy subjects (5 male, mean age = 37 ± 18 years), where an enlarged acquisition window of 166-220 ms was set 50 ms prior to the scout-derived trigger delay. Coronary visualization and sharpness scores were compared between the standard 120 ms window set at the trigger delay, and those reconstructed using a manually adjusted window. The proposed method using manual adjustment was able to recover delineation of five mid and distal right coronary artery regions that were otherwise not visible from the standard window, and the sharpness scores improved in all coronary regions using the proposed method. This paper demonstrates the feasibility of a whole-heart coronary imaging approach that allows interactive selection of any subset of the enlarged acquisition window for a tailored reconstruction for each branch region.


Development of selective agents targeting serotonin 5HT1A receptors with subnanomolar activities based on a coumarin core.
A series of 18 new 5-[3-(4-aryl-1-piperazinyl)propoxy]coumarin derivatives from the corresponding bromoalkyl derivatives have been designed and synthesized by us using a microwave-assisted protocol. Radioligand binding assays of this series of compounds as well as a previously synthesized series of 17 structurally-similar compounds showed that six systems have very high affinities to the 5-HT1A receptor (0.3-1.0 nM) and good selectivity against the 5-HT2A receptor. Molecular docking, structural studies and structure-activity relationship studies were used to gain more insight into the atomistic details of ligand binding and rationalize the obtained results.


[State of antiendotoxin defense during community-acquired pneumonia].
To assess the effect of systemic endotoxinemia and LPS-induced immune response on severity of community-acquired pneumonia (CAP). Clinico-immunological follow-up of 60 patients with CAP of different severity was conducted. LPS was quantitatively measured by ENDOSAFE ENDOCHROME (0.015-0.12 EU/ml), LBP--by Hbt Human LBP ELISA (min 1 ng/ml), IgG to core region of LPS--by Hbt EndoCAb ELISA (min 0.125 MU/ml) ("HyCult biotechnology", Netherlands). Level of LPS in blood was not increased, whereas there was increase of antiendotoxin proteins in patients with more severe CAP with Gram-negative or mixed bacterial flora identified. Correlation between levels of LBP, IgG to core-region of LPS, IL-6, TNF-alpha, IL-4 and immune response abnormalities during severe forms of disease was demonstrated. State of antiendotoxin immunity is an etiologic marker of CAP and its severity.


Impact of age on 30-day postoperative outcome of surgery for ruptured and unruptured intracranial aneurysms.
To assess in a retrospective analysis of a prospectively collected database, the impact of increased age on 30-day postoperative outcomes of surgery for intracranial aneurysms (ICAs). 721 adult patients who underwent surgery for ICA were identified in the 2006-2012 American College of Surgeons' National Surgical Quality Improvement Program. Baseline characteristics and 30-day outcomes were stratified by age: <50 years (n=221), 50-60 years (n=221), and >60 years (n=266). Patients <50 and 50-60 years old were propensity score-matched to those aged >60 years. Logistic regression was used to examine the relationship between increased age and surgical outcome. In unadjusted analyses, age <50 years was associated with fewer postoperative complications (OR=0.5, 95% CI 0.3 to 0.7) and lower mortality (OR=0.4, 95% CI 0.2 to 0.9) compared with those aged >60 years. Patients aged between 50 and 60 years were less likely to have complications (OR=0.6, 95% CI 0.4 to 0.8) in unadjusted analyses. Upon propensity score matching, covariate balance was achieved for all age strata. In adjusted analyses, patients <50 years (OR=0.4, 95% CI 0.2 to 0.7) and 50-60 years (OR=0.5, 95% CI 0.3 to 0.8) of age continued to have fewer complications than those aged >60. Age >60 is independently associated with 30-day postoperative morbidity in patients undergoing surgery for ICA. The results of this study suggest age >60 should be considered an a priori risk factor in surgical management of ICA, regardless of associated comorbidities often associated with increased age.


Inhibitory effects of D-amino acids on Staphylococcus aureus biofilm development.
Biofilms are communities of cells held together by a self-produced extracellular matrix typically consisting of protein, exopolysaccharide, and often DNA. A natural signal for biofilm disassembly in Bacillus subtilis is certain D-amino acids, which are incorporated into the peptidoglycan and trigger the release of the protein component of the matrix. D-amino acids also prevent biofilm formation by the related Gram-positive bacterium Staphylococcus aureus. Here we employed fluorescence microscopy and confocal laser scanning microscopy to investigate how D-amino acids prevent biofilm formation by S. aureus. We report that biofilm formation takes place in two stages, initial attachment to surfaces, resulting in small foci, and the subsequent growth of the foci into large aggregates. D-amino acids did not prevent the initial surface attachment of cells but blocked the subsequent growth of the foci into larger assemblies of cells. Using protein- and polysaccharide-specific stains, we have shown that D-amino acids inhibited the accumulation of the protein component of the matrix but had little effect on exopolysaccharide production and localization within the biofilm. We conclude that D-amino acids act in an analogous manner to prevent biofilm development in B. subtilis and S. aureus. Finally, to investigate the potential utility of D-amino acids in preventing device-related infections, we have shown that surfaces impregnated with D-amino acids were effective in preventing biofilm growth.


Meiotic Crossing Over in Maize Knob Heterochromatin.
There is ample evidence that crossing over is suppressed in heterochromatin associated with centromeres and nucleolus organizers (NORs). This characteristic has been attributed to all heterochromatin, but the generalization may not be justified. To investigate the relationship of crossing over to heterochromatin that is not associated with centromeres or NORs, we used a combination of fluorescence in situ hybridization of the maize 180-bp knob repeat to show the locations of knob heterochromatin and fluorescent immunolocalization of MLH1 protein and AFD1 protein to show the locations of MLH1 foci on maize synaptonemal complexes (SCs, pachytene chromosomes). MLH1 foci correspond to the location of recombination nodules (RNs) that mark sites of crossing over. We found that MLH1 foci occur at similar frequencies per unit length of SC in interstitial knobs and in the 1 µm segments of SC in euchromatin immediately to either side of interstitial knobs. These results indicate not only that crossing over occurs within knob heterochromatin, but also that crossing over is not suppressed in the context of SC length in maize knobs. However, because there is more DNA per unit length of SC in knobs compared to euchromatin, crossing over is suppressed (but not eliminated) in knobs in the context of DNA length compared to adjacent euchromatin.


Camera footage and identification testimony undermine the availability of exculpatory alibi evidence.
The present field experiment investigated how alibi witnesses react when confronted with camera footage or identification testimony that incriminates an innocent suspect. Under the pretext of a problem-solving study, pairs of participants (N = 109) and confederates worked on an individual task with a dividing wall obstructing their view of each other. When the mobile phone of the experimenter was missing from an adjacent room at the end of the session, all participants confirmed that the confederate had not left the room. After several days, participants returned to the lab for a second session. They were asked to confirm their corroboration, orally and in writing, after learning that the confederate either had been identified from a photograph or was present on camera footage. A control group received no evidence. In this second session, written (but not oral) alibi corroboration was weaker in the incriminating evidence conditions (47%) than the no-evidence condition (81%), as hypothesized. Unexpectedly, corroboration was equally strong in the camera and identification evidence conditions. As expected, alibi corroboration was stronger in session 1 than in session 2 for both camera (89% and 31-46%) and identification evidence conditions (86% and 31-49%). The current findings provide first evidence that camera footage and eyewitness identification testimony can bear on the availability of exculpatory alibi evidence in court and emphasize the need to document incidents of evidence contamination.


Light-emitting diode 415 nm in the treatment of inflammatory acne: an open-label, multicentric, pilot investigation.
The management of acne remains a challenge, with current therapies linked to significant side effects and patient non-compliance. Phototherapy using blue light has been proven in the treatment of acne vulgaris and offers the clinician an effective alternative. To determine the effect of narrowband light-emitting diode (LED) blue light in the reduction of inflammatory and non-inflammatory lesions in patients with mild to moderate acne and to evaluate patient tolerance of the therapy. Forty-five patients were treated with high-intensity pure blue light, 415 nm and 48 J/cm2, receiving two treatments of 20 minutes per week for a period of 4-8 weeks. Clinical assessment was performed at baseline, and 2, 4 and 8 weeks after treatment. A patient's therapeutic response was measured using a global improvement scoring system. The mean improvement score was 3.14 at 4 weeks and 2.90 at 8 weeks. Nine patients experienced complete clearing at 8 weeks. The treatment was well tolerated, with 50% of patients highly satisfied with the treatment. This open-label study suggests the therapeutic efficacy of high-intensity LED pure blue light in the treatment of acne vulgaris with no reported side effects.


Treatment strategies and survival of older breast cancer patients - an international comparison between the Netherlands and Ireland.
Forty percent of breast cancers occur among older patients. Unfortunately, there is a lack of evidence for treatment guidelines for older breast cancer patients. The aim of this study is to compare treatment strategy and relative survival for operable breast cancer in the elderly between The Netherlands and Ireland. From the Dutch and Irish national cancer registries, women aged ≥65 years with non-metastatic breast cancer were included (2001-2009). Proportions of patients receiving guideline-adherent locoregional treatment, endocrine therapy, and chemotherapy were calculated and compared between the countries by stage. Secondly, 5-year relative survival was calculated by stage and compared between countries. Overall, 41,055 patients from The Netherlands and 5,826 patients from Ireland were included. Overall, more patients received guideline-adherent locoregional treatment in The Netherlands, overall (80% vs. 68%, adjusted p<0.001), stage I (83% vs. 65%, p<0.001), stage II (80% vs. 74%, p<0.001) and stage III (74% vs. 57%, P<0.001) disease. On the other hand, more systemic treatment was provided in Ireland, where endocrine therapy was prescribed to 92% of hormone receptor-positive patients, compared to 59% in The Netherlands. In The Netherlands, only 6% received chemotherapy, as compared 24% in Ireland. But relative survival was poorer in Ireland (5 years relative survival 89% vs. 83%), especially in stage II (87% vs. 85%) and stage III (61% vs. 58%) patients. Treatment for older breast cancer patients differed significantly on all treatment modalities between The Netherlands and Ireland. More locoregional treatment was provided in The Netherlands, and more systemic therapy was provided in Ireland. Relative survival for Irish patients was worse than for their Dutch counterparts. This finding should be a strong recommendation to study breast cancer treatment and survival internationally, with the ultimate goal to equalize the survival rates for breast cancer patients across Europe.


Characterization of additional sewage treatment technologies: ecotoxicological effects and levels of selected pharmaceuticals, hormones and endocrine disruptors.
In the present study, two conventional (with and without sand filter) and four additional (moving bed biofilm reactor, ozone, moving bed biofilm reactor combined with ozone and a membrane bio reactor) treatment technologies were operated in small-scale at Hammarby Sjöstad sewage treatment plant, Stockholm, Sweden. The effluents were tested with five short-term (≤ 7 days exposure) ecotoxicological tests, and analyzed for a number of target analytes, comprising pharmaceuticals, natural hormones and industrial chemicals. Overall, the tested effluents generated few adverse effects at lower concentrations (< 50% sewage effluent), and no major differences were observed between any of the treatments. The effluent treated with the moving bed biofilm reactor resulted in the lowest effects in the ecotoxicological tests. The most efficient treatment technology with regard to the pharmaceutical residues was the ozone treatment, which however caused negative effects in some of the ecotoxicological tests.


Bromethalin poisoning in a raccoon (Procyon lotor): diagnostic considerations and relevance to nontarget wildlife.
Submission of a raccoon (Procyon lotor) for necropsy following exhaustion at a California wildlife care center revealed minimal gross pathologic changes and only mild vacuolar changes in the white matter of the brain. Turquoise granular material was noted in the gastrointestinal tract and was submitted for toxicological testing along with portions of the brain, liver, kidney, and mesenteric and perirenal adipose tissues. Testing of the turquoise material for 7 anticoagulant rodenticides, strychnine, 4-aminopyridine, starlicide, and salts revealed none of these compounds; however, desmethylbromethalin was detected by high-performance liquid chromatography-tandem mass spectrometry. Other tissues were subsequently analyzed; the mesenteric and perirenal adipose tissues contained desmethylbromethalin. Desmethylbromethalin is the active metabolite of bromethalin, uncouples oxidative phosphorylation, and results in cerebral edema. Bromethalin is a rodenticide that is visually indistinguishable from many other rodenticides, making identification of poisonings by appearance alone nearly impossible. Based on the pathological and toxicological findings, a diagnosis of bromethalin toxicosis was established. In cases of wildlife species with unknown deaths or inconsistent clinical signs with normal or minimal histological findings, bromethalin toxicosis should be considered as a differential. Adipose tissue is the tissue of choice and can be easily harvested from a live or deceased animal to help confirm or rule out bromethalin exposure or intoxication.


Network-based statistical comparison of citation topology of bibliographic databases.
Modern bibliographic databases provide the basis for scientific research and its evaluation. While their content and structure differ substantially, there exist only informal notions on their reliability. Here we compare the topological consistency of citation networks extracted from six popular bibliographic databases including Web of Science, CiteSeer and arXiv.org. The networks are assessed through a rich set of local and global graph statistics. We first reveal statistically significant inconsistencies between some of the databases with respect to individual statistics. For example, the introduced field bow-tie decomposition of DBLP Computer Science Bibliography substantially differs from the rest due to the coverage of the database, while the citation information within arXiv.org is the most exhaustive. Finally, we compare the databases over multiple graph statistics using the critical difference diagram. The citation topology of DBLP Computer Science Bibliography is the least consistent with the rest, while, not surprisingly, Web of Science is significantly more reliable from the perspective of consistency. This work can serve either as a reference for scholars in bibliometrics and scientometrics or a scientific evaluation guideline for governments and research agencies.


Pharmacological exploration of the sedative mechanism of hesperidin identified as the active principle of Citrus sinensis flowers.
The infusion of flowers of several species of Citrus genera is used as a sedative to treat insomnia in Mexican traditional medicine. The aims of this study were to investigate the sedative effect of different extracts of flowers of Citrus sinensis (L.) Osbeck (Rutaceae) and describe the pharmacological action mechanism of the sedative active compounds of this plant. The methanol and dichloromethane extracts, obtained from the flowers of Citrus sinensis (L.) Osbeck (Rutaceae), showed a dose-dependent sedative effect in the exploratory cylinder model in mice, with ED50 (ip) values of 47.04+/-12.03 mg/kg and 129.15+/-21.25 mg/kg, respectively. Hesperidin (ED50=11.34+/-2.48 mg/kg) was identified in the methanol extract as the sedative active principle of this plant. The pre-treatment with atropine (1 mg/kg I. P.), flumazenil (2 mg/kg I. P.), clonidine (0.01 mg/kg I. P.), isoproterenol (0.3 mg/kg I. P.), haloperidol (0.3 mg/kg I. P.), WAY 100 635 (3 mg/kg I. P.), P-chlorophenylalanine (250 mg/kg I. P., twice per day for 2 days), forskolin (3 mg/kg I. P.) and rolipram (0.173 mg/kg I. P.) did not modify the sedative effect of 30 mg/kg hesperidin. However, the sedative effect of this compound was potentiated by yohimbine (1.25 mg/kg I. P.) and buspirone (1 mg/kg I. P.), and reverted by pretreatment with aminophylline (30 mg/kg I. P.), caffeine (30 mg/kg I. P.) and several doses of 1,3-dimethyl-8-phenylxanthine (10, 30 and 54.7 mg/kg I. P.). These results suggest that adenosine receptors might be involved in the sedative action of hesperidin, identified as the active principle of the flowers of Citrus sinensis.


A rapid, accurate and robust UHPLC-MS/MS method for quantitative determination of BMS-927711, a CGRP receptor antagonist, in plasma in support of non-clinical toxicokinetic studies.
BMS-927711 is a calcitonin gene-related peptide (CGRP) receptor antagonist that is being developed for the treatment of migraine. A rapid, accurate and robust assay was developed and validated for the quantitation of BMS-927711 in rat, monkey, rabbit and mouse plasma using ultra high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). A simplified method screening strategy was utilized that included a liquid-liquid extraction (LLE) methodology and eleven LC columns (ten sub-2 μm UHPLC columns and one 2.6 μm HPLC column) for screening with emphasis on the removal of phospholipids, avoidance of metabolite interference and ruggedness of LC conditions. A stable isotope labeled [(13)C2, D4]-BMS-927711 was used as the internal standard, and 50 μL of plasma samples were used for extraction by automated LLE with methyl tert-butyl ether (MTBE) in 96-well format. Chromatographic separation was achieved with an isocratic elution and a gradient column wash on a Waters Acuity UPLC(®) BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with run time of 3.7 min. Positive electrospray ionization was performed using selected reaction monitoring (SRM) with transitions of m/z 535>256 for BMS-927711 and m/z 541>256 for [(13)C2, D4]-BMS-927711. The standard curve, which ranged from 3.00 to 3000 ng/mL for BMS-927711, was fitted to a 1/x(2) weighted linear regression model. The intra-assay precision was within 5.2% CV, inter-assay precision was within 5.9% CV, and the assay accuracy was within ±5.2% deviation (%Dev) of the nominal values in all the species. The stability of an N-carbamoyl glucuronide metabolite was carefully investigated, and the conversion of this metabolite to BMS-927711 was minimal and manageable without a stabilization procedure. The method was successfully applied to multiple non-clinical toxicokinetic studies in different species in support of the investigative new drug (IND) filing.


Filamentous white matter prion protein deposition is a distinctive feature of multiple inherited prion diseases.
Sporadic, inherited and acquired prion diseases show distinct histological patterns of abnormal prion protein (PrP) deposits. Many of the inherited prion diseases show striking histological patterns, which often associate with specific mutations. Most reports have focused on the pattern of PrP deposition in the cortical or cerebellar grey matter. We observed that the subcortical white matter in inherited prion diseases frequently contained filamentous depositions of abnormal PrP, and we have analysed by immunohistochemistry, immunofluorescence and electron microscopy 35 cases of inherited prion disease seen at the UK National Prion Clinic. We report here that filamentous PrP is abundantly deposited in myelinated fibres in inherited prion diseases, in particular in those with N-terminal mutations. It is possible that the presence of filamentous PrP is related to the pathogenesis of inherited forms, which is different from those sporadic and acquired forms.


The Potential Role of the NLRP3 Inflammasome as a Link between Mitochondrial Complex I Dysfunction and Inflammation in Bipolar Disorder.
Mitochondrial dysfunction and activation of the inflammatory system are two of the most consistently reported findings in bipolar disorder (BD). More specifically, altered levels of inflammatory cytokines and decreased levels of mitochondrial complex I subunits have been found in the brain and periphery of patients with BD, which could lead to increased production of mitochondrial reactive oxygen species (ROS). Recent studies have shown that mitochondrial production of ROS and inflammation may be closely linked through a redox sensor known as nod-like receptor pyrin domain-containing 3 (NLRP3). Upon sensing mitochondrial release of ROS, NLRP3 assembles the NLRP3 inflammasome, which releases caspase 1 to begin the inflammatory cascade. In this review, we discuss the potential role of the NLRP3 inflammasome as a link between complex I dysfunction and inflammation in BD and its therapeutic implications.


Anomalous ligand effect in gold(I)-catalyzed intramolecular hydroamination of alkynes.
We analyzed the ligand electronic effect in a gold(I)-catalyzed intramolecular alkyne hydroamination, through a DFT and charge-displacement function (CDF) study. We found that, in the presence of π-electron conjugation between the alkyne and the nucleophilic functionality, electron poor ligands modify the coordination mode and the geometric parameters of the substrate in such a way that, contrary to expectations, the activation barrier of the nucleophilic attack increases. This remarkable effect is due to the competition between alkyne activation and nucleophile deactivation. The general relevance of these findings is highlighted.


Pathophysiology and Treatment of Diabetic Cardiomyopathy and Heart Failure in Patients with Diabetes Mellitus.
There is a close relationship between diabetes mellitus and heart failure, and diabetes is an independent risk factor for heart failure. Diabetes and heart failure are linked by not only the complication of ischemic heart disease, but also by metabolic disorders such as glucose toxicity and lipotoxicity based on insulin resistance. Cardiac dysfunction in the absence of coronary artery disease, hypertension, and valvular disease is called diabetic cardiomyopathy. Diabetes-induced hyperglycemia and hyperinsulinemia lead to capillary damage, myocardial fibrosis, and myocardial hypertrophy with mitochondrial dysfunction. Lipotoxicity with extensive fat deposits or lipid droplets is observed on cardiomyocytes. Furthermore, increased oxidative stress and inflammation cause cardiac fibrosis and hypertrophy. Treatment with a sodium glucose cotransporter 2 (SGLT2) inhibitor is currently one of the most effective treatments for heart failure associated with diabetes. However, an effective treatment for lipotoxicity of the myocardium has not yet been established, and the establishment of an effective treatment is needed in the future. This review provides an overview of heart failure in diabetic patients for the clinical practice of clinicians.


Photo-oxygenation inhibits tau amyloid formation.
Tau amyloid formation and deposition are responsible for the onset of Alzheimer's disease. In particular, the seeding activity of the tau protein plays an important role in the spreading of tau pathology via its propagation in the human brain. Here we demonstrate that catalytic photo-oxygenation markedly suppresses tau seeding activity, resulting in the inhibition of amyloid formation, both in vitro and in cultured cells.


Taste-Masked Flucloxacillin Powder Part 2: Formulation Optimisation Using the Mixture Design Approach and Storage Stability.
Flucloxacillin is prescribed to treat skin infections but its highly bitter taste is poorly tolerated in children. This work describes the application of the D-optimal mixture experimental design to identify the optimal component ratio of flucloxacillin, Eudragit EPO and palmitic acid to prepare flucloxacillin taste-masked microparticles that would be stable to storage and would inhibit flucloxacillin release in the oral cavity while facilitating the total release of the flucloxacillin load in the lower gastrointestinal tract (GIT). The model predicted ratio was found to be very close to the stoichiometric equimolar component ratio, which supported our hypothesis that the ionic interactions among flucloxacillin, Eudragit EPO and palmitic acid underscore the polyelectrolyte complex formation in the flucloxacillin taste-masked microparticles. The excipient-drug interactions showed protective effects on the microparticle storage stability and minimised flucloxacillin release at 2 min in dissolution medium. These interactions had less influence on flucloxacillin release in the dissolution medium at 60 min. Storage temperature and relative humidity significantly affected the chemical stability of the microparticles. At the preferred storage conditions of ambient temperature under reduced RH of 23%, over 90% of the baseline drug load was retained in the microparticles at 12 months of storage.


Performance-based financing: the same is different.
Although it is increasingly acknowledged within the Performance-Based Financing (PBF) research community that PBF is more than just payments based on outputs verified for quality, this narrow definition of PBF is still very present in many studies and evaluations. This leads to missed opportunities, misunderstandings and an unhelpful debate. Therefore, we reinforce the claim that PBF should be viewed as a reform package focused on targeted services with many different aspects that go beyond the health worker level. Failing to acknowledge the importance of the different elements of PBF negatively influences the task of practitioners, researchers and policymakers alike. After making the case for this wider definition, we propose three research pathways (describing, understanding and framing PBF) and give a short and tentative starting point for future research, leaving the floor open for more in-depth discussions. From these three vantage points it appears that when it comes to PBF 'the same is different'. Notwithstanding the increased complexity due to the use of the wider definition, progress on these three different research pathways will strongly improve our knowledge, lead to better adapted PBF programs and create a more nuanced debate on PBF.


CCDC6: the identity of a protein known to be partner in fusion.
Coiled Coil Domain Containing 6 gene, CCDC6, was initially isolated as part of a tumorigenic DNA originated by the fusion of CCDC6 with the tyrosine kinase of RET receptor, following a paracentric inversion of chromosome 10. For a long time, CCDC6 has been considered as an accidental partner of the RET protooncogene, providing the promoter and the first 101 aa necessary for the constitutive activation of the oncogenic Tyrosine Kinase (TK) RET in thyroid cells. With the advent of more refined diagnostic tools and bioinformatic algorithms, an exponential growth in fusion genes discoveries has allowed the identification of CCDC6 as partner of genes other than RET in different tumor types. CCDC6 gene product has a proper role in sustaining the DNA damage checkpoints in response to DNA damage. The inactivation of CCDC6 secondary to chromosomal rearrangements or gene mutations could enhance tumor progression by impairing the apoptotic response upon the DNA damage exposure, contributing to the generation of radio- and chemoresistance. Preclinical studies indicate that the attenuation of CCDC6 in cancer, while conferring a resistance to cisplatinum, sensitizes the cancer cells to the small molecule inhibitors of Poly (ADP-ribose) polymerase (PARP1/2) with a synthetic lethal effect. Several CCDC6 mutations and gene rearrangements have been described so far in different types of cancer and CCDC6 may represent a possible predictive biomarker of tumor resistance to the conventional anticancer treatments. Nevertheless, the detection of a CCDC6 impairment in cancer patients may help to select, in future clinical trials, those patients who could benefit of PARP-inhibitors treatment alone or in combination with other treatments.


NETWORK MODELLING OF TOPOLOGICAL DOMAINS USING HI-C DATA.
Chromosome conformation capture experiments such as Hi-C are used to map the three-dimensional spatial organization of genomes. One specific feature of the 3D organization is known as topologically associating domains (TADs), which are densely interacting, contiguous chromatin regions playing important roles in regulating gene expression. A few algorithms have been proposed to detect TADs. In particular, the structure of Hi-C data naturally inspires application of community detection methods. However, one of the drawbacks of community detection is that most methods take exchangeability of the nodes in the network for granted; whereas the nodes in this case, that is, the positions on the chromosomes, are not exchangeable. We propose a network model for detecting TADs using Hi-C data that takes into account this nonexchangeability. in addition, our model explicitly makes use of cell-type specific CTCF binding sites as biological covariates and can be used to identify conserved TADs across multiple cell types. The model leads to a likelihood objective that can be efficiently optimized via relaxation. We also prove that when suitably initialized, this model finds the underlying TAD structure with high probability. using simulated data, we show the advantages of our method and the caveats of popular community detection methods, such as spectral clustering, in this application. Applying our method to real Hi-C data, we demonstrate the domains identified have desirable epigenetic features and compare them across different cell types.


Spin coupling in engineered atomic structures.
We used a scanning tunneling microscope to probe the interactions between spins in individual atomic-scale magnetic structures. Linear chains of 1 to 10 manganese atoms were assembled one atom at a time on a thin insulating layer, and the spin excitation spectra of these structures were measured with inelastic electron tunneling spectroscopy. We observed excitations of the coupled atomic spins that can change both the total spin and its orientation. Comparison with a model spin-interaction Hamiltonian yielded the collective spin configuration and the strength of the coupling between the atomic spins.


Chemotherapy in elderly patients with nonsmall cell lung cancer.
Using chemotherapy in elderly nonsmall cell lung cancer (NSCLC) patients is often challenging given concerns of treatment-related toxicity. However, data have demonstrated that chemotherapy can lead to improved survival in this age group. In this review, we summarize existing data and discuss the role of chemotherapy in elderly patients with localized, locally advanced, and metastatic NSCLC. Clear evidence-based guidelines for chemotherapy management in elderly patients is lacking given the limited prospective data available. However, there are more clinical trials investigating optimal chemotherapy agents and dosing schedules specific to the elderly. Comprehensive geriatric assessment-directed interventions are also being prospectively investigated to improve treatment selection for elderly patients. Chronological age should not be a limiting factor for chemotherapy use in elderly NSCLC patients. Several studies have demonstrated similar survival benefits than in younger patients when chemotherapy is given as adjuvant treatment for localized disease; part of definitive treatment with radiation in locally advanced disease; and palliative treatment for advanced NSCLC, however, at the cost of greater toxicity. Tolerability of chemotherapy in this heterogeneous group can be difficult to predict. Therefore, therapeutic decisions should be individualized based on performance status. Comprehensive geriatric assessment should be used to supplement performance status measures to minimize both under and overtreatment.


Microproteinuria and long-term prognosis with respect to renal function and survival in normotensive and hypertensive women--a 24-year follow-up of a representative population sample of women in Gothenburg, Sweden.
This study aimed to assess albuminuria and subclinical proteinuria, their association with hypertension and their role as predictors of hypertension, impaired renal function and mortality. A baseline population study comprising 1462 women in five different age groups in Gothenburg, Sweden, was carried out in 1968-69. Comprehensive clinical examinations and laboratory tests were performed, including blood pressure measurement and an Albustix test. A systematic subsample of women additionally collected a 24 h urine sample for quantitative protein analysis. Values of urinary protein (u-protein) excretion between 80 and 300 mg/24 h were defined as microproteinuria. The results described in this paper are based on a 24-year follow-up. The baseline Albustix test was positive in 6.8% of 1458 women, from whom a urine sample was obtained. Of 741 baseline urine collections for u-protein excretion, 16.9% were in the microproteinuric range (80-300 mg/24 h), 1.1% in the macroproteinuric range (> 300 mg/24 h) and 82.1% in the normoproteinuric range (< 80 mg/24 h). Hypertension was more common in Albustix-positive women than in those with negative Albustix, and hypertension was also more prevalent in women with microproteinuria than in women with normoproteinuria. Neither positive Albustix nor microproteinuria was related to later renal impairment. Hypertension was associated with increased mortality in both Albustix-positive and Albustix-negative women, and in women with both normoproteinuria and microproteinuria at baseline. The mortality ratio during the follow-up period was, however, not significantly influenced by positive Albustix or by microproteinuria at baseline, in either hypertensive or non-hypertensive women. This study demonstrated that both a positive Albustix test and microproteinuria were associated with hypertension. Hypertension at baseline increased the risk for death during the follow-up period, while neither albuminuria, defined as a positive Albustix test, nor microproteinuria was associated with an impaired long-term prognosis with respect to renal function or survival in this cohort of Swedish middle-aged women during 24 years of follow-up. Microproteinuria in otherwise healthy normotensive or hypertensive women does not appear to impair the long-term prognosis.


Natural genetic variation underlying differences in Peromyscus repetitive and social/aggressive behaviors.
Peromyscus maniculatus (BW) and P. polionotus (PO) are interfertile North American species that differ in many characteristics. For example, PO exhibit monogamy and BW animals are susceptible to repetitive behaviors and thus a model for neurobehavioral disorders such as Autism. We analyzed these two stocks as well as their hybrids, a BW Y(PO) consomic line (previously shown to alter glucose homeostasis) and a natural P. maniculatus agouti variant (A(Nb) = wide band agouti). We show that PO animals engage in far less repetitive behavior than BW animals, that this trait is dominant, and that trait distribution in both species is bi-modal. The A(Nb) allele also reduces such behaviors, particularly in females. PO, F1, and A(Nb) animals all dig significantly more than BW. Increased self-grooming is also a PO dominant trait, and there is a bimodal trait distribution in all groups except BW. The inter-stock differences in self-grooming are greater between males, and the consomic data suggest the Y chromosome plays a role. The monogamous PO animals engage in more social behavior than BW; hybrid animals exhibit intermediate levels. Surprisingly, A(Nb) animals are also more social than BW animals, although A(Nb) interactions led to aggressive interactions at higher levels than any other group. PO animals exhibited the lowest incidence of aggressive behaviors, while the hybrids exhibited BW levels. Thus this group exhibits natural, genetically tractable variation in several biomedically relevant traits.


[Appropriate care in the last phase of life].
To identify the types of care that patients and relatives perceive as appropriate or inappropriate in the last phase of life. Questionnaire survey. By means of an internet questionnaire with open questions, we asked patients and relatives about their experiences of care in the last phase of life and if they found it appropriate or inappropriate, and why. We recruited via e-mails, Twitter, Facebook, newsletters and links on homepages of patient organisations. Members of a Dutch large organisation for older people (Unie KBO) received the questionnaire on paper. Forty-five patients and 547 relatives described received one or more cases. We found 429 cases of appropriate care and 309 different cases of inappropriate care. We identified five common dimensions of appropriate and inappropriate care, i.e., supportive care, treatment decisions, location of care, following the patient's wishes and communication. In the cases of appropriate care, all five dimensions were regularly mentioned; supportive care being most-frequently mentioned. In cases of inappropriate care, the dimensions treatment decisions, supportive care and communication were most often mentioned. Descriptions of appropriate care more frequently concerned patients with cancer, and descriptions of inappropriate care more frequently concerned patients with other somatic conditions or psychiatric conditions. Nurses or carers and general practitioners were more often mentioned in descriptions of appropriate care, and medical specialists more often mentioned in descriptions of inappropriate care. Appropriate care appears to be a wide-ranging term covering supportive care, treatment decisions, communication, following the patient's wishes, and the location where care is provided. Inappropriate treatment decisions and communication problems are the main threats to appropriate care in the last phase of life.


A novel calmodulin-like protein gene in rice which has an unusual prolonged C-terminal sequence carrying a putative prenylation site.
A rice cDNA encoding a novel calmodulin-like protein was identified. It has 38 additional amino acids at the C-terminus of a complete, typical calmodulin (CaM) sequence of 149 amino acids. The four C-terminal amino acid residues form a CAAL motif which could be a site for protein prenylation and may subsequently cause the protein to become membrane associated. RT-PCR analysis confirmed that such a combined protein gene truly exists in rice. Sequence analysis of its genomic counterpart showed that there is an intron located at junction of the normal CaM sequence and the 38 C-terminal amino acids. This introduces a potential stop codon for normal CaM if an alternative splicing mechanism is involved. Southern blot analysis of rice genomic DNA revealed that there is only one locus for this gene. The northern blot analysis showed that this gene is highly expressed in rice roots, shoots and flowers. The distribution of this protein demonstrates the functional importance of this novel CaM-like protein in rice.


Feasibility studies in spheronization and scale-up of ibuprofen microparticulates using the rotor disk fluid-bed technology.
The aim of this study was to develop spheronized microparticulates as a drug delivery system using the 1-step closed rotor disk fluid-bed technology, and to scale up the batch spheronization process. Ibuprofen was used as the model drug and microcrystalline cellulose/sodium carboxymethyl cellulose hydrocolloid (Avicel(R) RC-581 or CL-611) was present as the diluent/binder. The mixture, in 1:1 ratio, was blended with and without 1% sodium lauryl sulfate (SLS) and spheronized with the rotor disk insert, using either water or hydroxypropylmethyl cellulose (HPMC) as binder. Fluid-bed machines (Vector/Freund Flo-Coater model) FLM-1 (with 9-inch rotor insert for 0.75 kg) and FLM-15 (with a 12-inch and 19-inch rotor inserts for 1 kg and 5, 10 kg, respectively) were used. The critical process parameters included inlet air temperature, rotor disk speed and configuration, air flow, and rate of binder application. The 1 kg batch containing SLS that was made with 12-inch smooth stainless steel or waffle teflon plates rotating at 500 rpm had desirable characteristics. The sphericity values were 0.88 and 0.91, with percent yield of 85.4 and 91.2 and drug content values of 94.47% and 91.44%, respectively. The spheroids showed good flow properties with respective rapid drug release (Q20 = 83.27 and 91.75). No difference was seen in the Avicel RC-581 and CL-611. Based on the 1 kg data, Avicel RC-581 and smooth stainless steel and waffle teflon plates (12 inch and 19 inch), the batch was scaled up to 5 and 10 kg. The scale-up parameters included rotor speed (124 -300 rpm) and spray rate (90-140 g/min). The scale-up batches had similar flow characteristics, release rate, and size distribution. The geometric mean diameter increased as batch size increased, and slightly bigger spheroids were obtained using the waffle teflon plate. Ibuprofen spheres with very good physical characteristics were developed using the rotor disk fluid-bed technology, a 1-step closed process that did not require additional unit processes.


Gelatin promotes rapid restoration of the blood brain barrier after acute brain injury.
Gelatin coating of brain implants is known to provide considerable benefits in terms of reduced inflammatory sequalae and long-term neuroprotective effects. However, the mechanisms for gelatin's protective role in brain injury are still unknown. To address this question, cellular and molecular markers were studied with quantitative immunohistochemical microscopy at acute (<2hours, 1, 3days), intermediate (1-2 weeks) and long-term time points (6 weeks) after transient insertion of stainless steel needles into female rat cortex cerebri with or without gelatin coating. Compared to non-coated controls, injuries caused by gelatin coated needles showed a significantly faster resolution of post-stab bleeding/leakage and differential effects on different groups of microglia cells. While similar levels of matrix metalloproteinase (MMP-2 and MMP-9, two gelatinases) was found for coated and noncoated needle stabs during the first week, markedly increased levels of both MMPs was seen for gelatin-coated but not non-coated needle stabs after 2weeks. Neuronal populations and activated astrocytes were largely unaffected. In conclusion, the beneficial effects of gelatin may be the combined results of faster healing of the blood brain barrier curtailing leakage of blood borne molecules/cells into brain parenchyma and to a modulation of the microglial population response favoring restitution of the injured tissue. These findings present an important therapeutic potential for gelatin coatings in various disease, injury and surgical conditions. The neural interfaces field holds great promise to enable elucidation of neural information processing and to develop new implantable devices for stimulation based therapy. Currently, this field is struggling to find solutions for reducing tissue reactions to implanted micro and nanotechnology. Prior studies have recently shown that gelatin coatings lower activation of digestive microglia and mitigate the ubiquitous loss of neurons adjacent to implanted probes, both of which impede implant function. The underlying mechanisms remain to be elucidated, however. Our findings demonstrate for the first time that gelatin has a significant effect on the BBB by promoting rapid restoration of integrity after injury. Moreover, gelatin alters microglia phenotypes and modulates gelatinase activity for up to 2weeks favoring anti-inflammation and restoration of the tissue. Given the key importance of the BBB for normal brain functions, we believe our findings have substantial significance and will be highly interesting to researchers in the biomaterial field.


Serum activin A and B, and follistatin in critically ill patients with influenza A(H1N1) infection.
Activin A and its binding protein follistatin (FS) are increased in inflammatory disorders and sepsis. Overexpression of activin A in the lung causes similar histopathological changes as acute respiratory distress syndrome (ARDS). ARDS and severe respiratory failure are complications of influenza A(H1N1) infection. Interleukin 6 (IL-6), which in experimental studies increases after activin A release, is known to be related to the severity of H1N1 infection. Our aim was to evaluate the levels of activin A, activin B, FS, IL-6 and IL-10 and their association with the severity of respiratory failure in critically ill H1N1 patients. A substudy of a prospective, observational cohort of H1N1 patients in Finnish intensive care units (ICU). Clinical information was recorded during ICU treatment, and serum activin A, activin B, FS, IL-6 and IL-10 were measured at admission to ICU and on days 2 and 7. Blood samples from 29 patients were analysed. At the time of admission to intensive care unit, elevated serum levels above the normal range for respective age group and sex were observed in 44% for activin A, 57% for activin B, and 39% for FS. In 13 of the 29 patients, serial samples at all time points were available and in these the highest activin A, activin B and FS were above the normal range in 85%, 100% and 46% of the patients, respectively. No difference in baseline or highest activin A or activin B was found in patients with or without acute lung injury (ALI) or ARDS (P > 0.05 for all). Peak levels of IL-6 were significantly elevated in ALI/ARDS patients. Peak activin A and activin A/FS were associated with ventilatory support free-days, severity of acute illness and length of ICU stay (P < 0.05 for all). Higher than normal values of these proteins were common in patients with H1N1 infection but we found no association with the severity of their respiratory failure.


Alpha-ketoglutarate affects murine embryo development through metabolic and epigenetic modulations.
α-Ketoglutarate (α-KG) is an intermediary metabolite in the tricarboxylic acid (TCA) cycle and functions to inhibit ATPase and maintain the pluripotency of embryonic stem cells (ESCs); however, little is known regarding the effects of α-KG on the development of preimplantation embryos. Herein, we report that α-KG (150 μM) treatment significantly promoted the blastocyst rate, the number of inner cell mass (ICM) cells and foetal growth after embryo transfer. Mechanistic studies revealed two important pathways involved in the α-KG effects on embryo development. First, α-KG modulates mitochondria function by inducing relatively low ATP production without modification of mitochondrial copy number. The relatively low energy metabolism preserves the pluripotency and competence of the ICM. Second, α-KG modifies epigenetics in embryos cultured in vitro by affecting the activity of the DNA demethylation enzyme TET and the DNA methylation gene Dnmt3a to increase the ratio of 5hmC/5mC ratio. Elevation of the 5hmC/5mC ratio not only promotes the pluripotency of the ICM but also leads to a methylation level in an in vitro embryo close to that in an in vivo embryo. All these functions of α-KG collectively contribute to an increase in the number of ICM cells, leading to greater adaptation of cultured embryos to in vitro conditions and promoting foetal growth after embryo transfer. Our findings provide basic knowledge regarding the mechanisms by which α-KG affects embryo development and cell differentiation.


Incorporating RNA-seq data into the zebrafish Ensembl genebuild.
Ensembl gene annotation provides a comprehensive catalog of transcripts aligned to the reference sequence. It relies on publicly available species-specific and orthologous transcripts plus their inferred protein sequence. The accuracy of gene models is improved by increasing the species-specific component that can be cost-effectively achieved using RNA-seq. Two zebrafish gene annotations are presented in Ensembl version 62 built on the Zv9 reference sequence. Firstly, RNA-seq data from five tissues and seven developmental stages were assembled into 25,748 gene models. A 3'-end capture and sequencing protocol was developed to predict the 3' ends of transcripts, and 46.1% of the original models were subsequently refined. Secondly, a standard Ensembl genebuild, incorporating carefully filtered elements from the RNA-seq-only build, followed by a merge with the manually curated VEGA database, produced a comprehensive annotation of 26,152 genes represented by 51,569 transcripts. The RNA-seq-only and the Ensembl/VEGA genebuilds contribute contrasting elements to the final genebuild. The RNA-seq genebuild was used to adjust intron/exon boundaries of orthologous defined models, confirm their expression, and improve 3' untranslated regions. Importantly, the inferred protein alignments within the Ensembl genebuild conferred proof of model contiguity for the RNA-seq models. The zebrafish gene annotation has been enhanced by the incorporation of RNA-seq data and the pipeline will be used for other organisms. Organisms with little species-specific cDNA data will generally benefit the most.


Inhalation of electronic cigarette aerosol induces reflex bronchoconstriction by activation of vagal bronchopulmonary C-fibers.
The electronic cigarette (e-cig) has been suggested as a safer alternative to tobacco cigarettes. However, the health effects of e-cigs on the airways have not been fully investigated. Nicotine, the primary chemical constituent of the e-cig aerosol, has been shown to stimulate vagal bronchopulmonary C-fiber sensory nerves, which upon activation can elicit vigorous pulmonary defense reflexes, including airway constriction. In this study, we investigated the bronchomotor response to e-cig inhalation challenge in anesthetized guinea pigs and the mechanisms involved in regulating these responses. Our results showed that delivery of a single puff of e-cig aerosol into the lung triggered immediately a transient bronchoconstriction that sustained for >2 min. The increase in airway resistance was almost completely abolished by a pretreatment with either intravenous injection of atropine or inhalation of aerosolized lidocaine, suggesting that the bronchoconstriction was elicited by cholinergic reflex mechanism and stimulation of airway sensory nerves was probably involved. Indeed, electrophysiological recording further confirmed that inhalation of e-cig aerosol exerted a pronounced stimulatory effect on vagal bronchopulmonary C-fibers. These effects on airway resistance and bronchopulmonary C-fiber activity were absent when the e-cig aerosol containing zero nicotine was inhaled, indicating a critical role of nicotine. Furthermore, a pretreatment with nicotinic acetylcholine receptor antagonists by inhalation completely prevented the airway constriction evoked by e-cig aerosol inhalation. In conclusion, inhalation of a single puff of e-cig aerosol caused a transient bronchoconstriction that was mediated through cholinergic reflex and triggered by a stimulatory effect of nicotine on vagal bronchopulmonary C-fiber afferents.


Dynamics of foam flow in a rock fracture: Effects of aperture variation on apparent shear viscosity and bubble morphology.
There has recently been renewed interest in understanding the physics of foam flow in permeable media. As for Newtonian flows in fractures, the heterogeneity of local apertures in natural fractures is expected to strongly impact the spatial distribution of foam flow. Although several experimental studies have been previously performed to study foam flow in fractured media, none of them has specifically addressed that impact for parallel flow in a realistic fracture geometry and its consequences for the foam's in situ shear viscosity and bubble morphologies. To do so, a comprehensive series of single-phase experiments have been performed by injecting pre-generated foams with six different qualities at a constant flow rate through a replica of a Vosges sandstone fracture of well-characterized aperture map. These measurements were compared to measurements obtained in a Hele-Shaw (i.e., smooth) fracture of identical hydraulic aperture. The results show that fracture wall roughness strongly increases the foam's apparent viscosity and shear rate. Moreover, foam bubbles traveling in regions of larger aperture exhibit larger velocity, size, a higher coarsening rate, and are subjected to a higher shear rate. This study also presents the first in situ measurement of foam bubbles velocities in fracture geometry, and provides hints towards measuring the in situ rheology of foam in a rough fracture from the velocity maps, for various imposed mean flow rates. These findings echo the necessity of considering fracture wall when predicting the pressure drop through the fracture and the effective viscosity, as well as in situ rheology, of the foam.


Resolving an apparent discrepancy between theory and experiment: spin-spin coupling constants for FCCF.
Ab initio equation of motion coupled cluster singles and doubles (EOM-CCSD) and second-order polarization propagator approximation (SOPPA) calculations have been performed to evaluate spin-spin coupling constants for FCCF (difluoroethyne). The computed EOM-CCSD value of (3)J(F-F) obtained at the experimental geometry of this molecule supports the previously reported experimental value of 2.1 Hz, thereby resolving an apparent discrepancy between theory and experiment. This coupling constant exhibits a strong dependence on the C-C and C-F distances, and its small positive value results from a sensitive balance of paramagnetic spin-orbit (PSO) and spin-dipole (SD) terms. The three other unique FCCF coupling constants (1)J(C-C), (1)J(C-F), and (2)J(C-F) have also been reported and compared with experimental data. While (1)J(C-F) is in agreement with experiment, the computed value of (2)J(C-F) is larger than our estimate of the experimental coupling constant.


Anti-idiotypic vaccination in the treatment of low-grade B-cell lymphoma.
Patients with B-cell lymphoma can be induced to mount a specific immune response against the individual idiotypic determinants expressed in their tumor cells. This form of active immunotherapy is now under evaluation in the clinical setting. We evaluated the feasibility and effectiveness of this kind of immunotherapy in a group of patients with low-grade lymphoma, which included two cases of bi/triclonal lymphoma. Nine patients with a histopathologic diagnosis of follicular non-Hodgkin's (NHL) low-grade B-cell lymphoma were initially selected for this disease-free survival study. Idiotypic proteins were recovered by somatic fusion of the tumor cells and their identity with the tumor idiotype determined by molecular methods. The patients received the vaccine consisting of their tumor Ig protein coupled to keyhole limpet hemocyanine and were observed for toxicity, anti-idiotypic immune response, clinical outcome and circulating t(14;18)+ tumor cells. The median duration of follow-up was 40 (10-64) months from the initiation of immunotherapy. Tumor regression was detected in two patients. No tumor progression was observed in the other patients. Eight patients generated specific anti-idiotypic antibodies and 3 out of five were cleared of circulating t(14;18)+ cells. Induction of tumor-specific anti-idiotypic immune responses may be of benefit to patients affected by low-grade B-cell NHL. Our results are in line with those previously reported and call attention to the issue of tumor clonality in this kind of treatment.


Impact of anthropometric parameters on outcomes in Asians with metabolic dysfunction-associated fatty liver disease.
We examined the incidence and predictors of clinical outcomes in metabolic dysfunction-associated fatty liver disease (MAFLD), focusing on anthropometric parameters. Adult patients with MAFLD were identified in nationwide databases and a hospital cohort. Primary endpoints were atherosclerotic cardiovascular disease (ASCVD) and advanced fibrosis. Logistic and Cox regression analyses were used to analyse the association between anthropometric parameters and endpoints. In total, 4407 of 15 256 (28.9%) and 6274 of 25 784 subjects (24.3%) had MAFLD in the nationwide database; of these, 403 (9.2%) and 437 (7.0%) subjects were of lean/normal weight, respectively. Compared to the overweight/obese group, the lean/normal weight group had a significantly lower muscle mass (15.0 vs. 18.9 kg) and handgrip strength (31.9 vs. 35.1 kg) and had a higher ASCVD risk (9.0% vs. 6.3% and 15.9% vs. 8.5%; Ps < 0.001). Sarcopenia (odds ratio [OR], 6.66; 95% confidence interval [CI], 1.79-24.80) and handgrip strength (OR, 0.92; 95% CI, 0.86-0.97; Ps = 0.005) were associated with the ASCVD risk in the lean/normal weight group. In a hospital cohort (n = 1363), the ASCVD risk was significantly higher in the lean/normal weight group than in the overweight/obese group (median follow-up, 39.1 months). Muscle mass was inversely correlated with the ASCVD risk (hazard ratio [HR], 0.72; 95% CI, 0.56-0.94), while visceral adiposity was associated with advanced fibrosis (HR, 1.36; 95% CI, 1.10-1.69; Ps < 0.05). Muscle mass/strength was significantly associated with the ASCVD risk in patients with MAFLD. Visceral adiposity was an independent predictor of advanced fibrosis.


Risk of disability pension for patients diagnosed with haematological malignancies: a register-based cohort study.
Patients with haematological malignancies are at increased risk of experiencing work-related problems. The aims of this study were to compare the risk of disability pension (DP) among patients diagnosed with eight subtypes of haematological malignancies to a reference cohort, and to determine if relative risks differ between these subtypes; to evaluate the influence of socioeconomic factors, demographic factors, and clinical factors on the risk of DP; and to investigate if these associations differ between the reference cohort and the patient cohort. We combined data from national registers on Danish patients diagnosed with haematological malignancies between 2000 and 2007 and a reference cohort without a history of these diseases. A total of 3194 patients and 28 627 reference individuals were followed until DP, emigration, old age pension or anticipatory pension, death or 26 February 2012, whichever came first. A total of 550 (17%) patients and 1511 (5%) reference individuals were granted DP. Age- and gender-adjusted relative risks differed significantly between the subgroups of haematological malignancies and ranged from 2.64 (95% CI 1.84-3.78) for patients with Hodgkin lymphoma to 12.53 (95% CI 10.57-14.85) for patients with multiple myeloma. In the patient cohort we found that gender, age, comorbidity, ethnicity, educational level, household income, history of long-term sick leave, and need of treatment with anxiolytics or antidepressants after diagnosis were associated with receiving DP. However, most of these associations were stronger in the reference cohort. All eight subtypes of haematological malignancies were associated with an increased risk of DP compared to the reference cohort. The relative risks differed according to subtype, and patients with multiple myeloma had the highest risk of DP. Furthermore, most socioeconomic, demographic and clinical factors had a stronger impact on the risk of DP in the reference cohort than in the patient cohort.


[Role of perforating veins in occurrence of varicose disease and its recurrences after surgical interventions].
The blood horizontal reflux via insufficient perforating veins (PV) from the deep venous system into subcutaneous one is considered an important factor of formation of chronic venous insufficiency in varicose disease (VD) and its recurrences (VDR). The role of PV in a VD progressing and in a VDR postoperatively in 774 patients, operated on for primary VD and 243 - VDR, is discussed. While determining significance of the blood horizontal reflux and substantiation of necessity for the PV ligation there must be taken into account the data of the colour duplex scanning concerning their valve apparatus state and clinical estimation of the disease severity.


The monoclonal antibody TER-119 recognizes a molecule associated with glycophorin A and specifically marks the late stages of murine erythroid lineage.
The antigen specificity of a rat monoclonal antibody TER-119 was investigated. In adult mice, TER-119 reacted with mature erythrocytes, 20-25% of bone marrow cells and 2-3% of spleen cells but not with thymocytes nor lymph node cells. In fetal haematopoietic tissues, 30-40% of d 10 yolk sac cells, 80-90% of d 14 fetal liver cells and 40-50% of newborn liver cells were reactive with TER-119. TER-119+ cells in adult bone marrow expressed significant levels of CD45 but not myeloid (Mac-1, Gr-1) or B-cell (B220) markers. Morphological examination and haematopoietic colony-forming assays for isolated TER-119+ cells revealed that TER-119 reacts with erythroid cells at differentiation stages from early proerythroblast to mature erythrocyte, but not with cells showing typical erythroid blast-forming unit (BFU-E) and erythroid colony-forming unit (CFU-E) activities. Erythroleukaemia cell lines do not express the TER-119 antigen even after stimulation with dimethylsulphoxide. TER-119 immunoprecipitated protein bands with molecular masses of 110 kDa, 60 kDa, 52 kDa and 32 kDa from erythrocyte membrane, whereas only a 52-kDa band was detected by TER-119 in Western blot analysis. Further molecular and cellular analyses indicated that the TER-119 antigen is a molecule associated with cell-surface glycophorin A but not with glycophorin A itself.


Evidence-based treatment of early-stage melanoma.
The vast majority of patients with newly diagnosed melanoma are "early-stage" - ie, clinically localized to the primary cutaneous site. For such patients, surgery represents the mainstay of treatment; current standards of surgical management have evolved based on evidence obtained from randomized trials, large multi-center and single institutional databases, and consensus panels. Treatment strategies include wide excision of the primary site with margins dictated by important biological features such as Breslow tumor thickness, and sentinel node biopsy to surgically evaluate the regional nodal basins at risk. Sentinel node biopsy has become an important component of the initial management of many of these patients for accurate staging of regional lymph nodes, as well as enhanced regional disease control and improved survival in the patients with microscopically involved nodes. Evidence is presented that supports the rational use of these surgical strategies and reconciles some of the controversies that have emerged. The degree to which established national treatment guidelines are followed and the consequences of non-compliance with these guidelines have become the focus of recent clinical investigations; results of these efforts are also discussed.


Analogues of 8-azaguanosine.
Two routes for the synthesis of 6-substtituted 8-azaguanosine analogues are described. 2,5,6-Triamino-4(3H)-pyrimidinethione (1) was converted by methylation, nitrosation, and acetylation to -n-acetyl-7-(methylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-5-amine (5). The reaction of 5 with 2,3,5-tri-O-acetyl-D-ribofuranosyl chloride gave a mixture of the 7-, 8-, and 9-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)-8-azapurines 4a-c which was converted to 8-azaguanosine (7c) and the corresponding 7- and 8-substituted isomers 7a and 7b. 4a-c were also converted with NaOMe to 6-O-methyl-8-azaguanosine (8c) and to the corresponding 7- and 8-substituted isomers 8a and 8b. The preferred route, however, to 6-substituted 8-azaguanosine analogues is an unambigous synthesis through N2-acetyl-6-(benzylthio)-N4-(2,3-O-isopropylidene-beta-D-ribofuranosyl)-5-nitro-2,4-pyrimidinediamine (13), prepared from the reaction of the chloropyrimidine 10 with the aminoribose 11. Catalytic hydrogenation of 13 gave the aminopyrimidine 14, which was converted with nitrous acid to the nucleoside beta-20. Treatment of beta-20 with dilute acid gave 7-(benzylthio)-3-beta-D-ribofuranosyl-3H-1,2,3-triazolo[4,5-d]pyrimidin-5-amine (19). Replacement of the benzylthio group of 19 with various nucleophilic reagents gave 8-aza-6-thioguanosine 17 and analogues 8c, 15, and 16. The thione 17 rearranges in aqueous solution to the thiasiazolopyrimidine 21. The parent [1,2,3]thiadiazolo[5,4-d]pyrimidine-5,7-diamine (24a) was prepared by nitrosation of the triaminopyrimidine (23a). Rearrangement of 24a in the presence of base gave a high yield of the thione 25a which could be rearranged with heat to 24a. Compounds 8a-c, 15-19, 24a, and 25 a were evaluated in the L1210 mouse leukemia screen- Only one compound, 8c, showed high cytotoxicity and borderline L1210 activity resulting from its en,ymatic conversion to 8-azaguanosine.


Oxidative Stress, Ocular Disease and Diabetes Retinopathy.
Oxidative stress is caused by free radicals or oxidant productions, including lipid peroxidation, protein modification, DNA damage and apoptosis or cell death and results in cellular degeneration and neurodegeneration from damage to macromolecules. Accumulation of the DNA damage (8HOdG) products and the end products of LPO (including aldehyde, diene, triene conjugates and Schiff's bases) were noted in the research studies. Significantly higher levels of these products in comparison with the controls were observed. Oxidative stress induced changes to ocular cells and tissues. Typical changes include ECM accumulation, cell dysfunction, cell death, advanced senescence, disarrangement or rearrangement of the cytoskeleton and released inflammatory cytokines. It is involved in ocular diseases, including keratoconus, Fuchs endothelial corneal dystrophy, and granular corneal dystrophy type 2, cataract, age-related macular degeneration, primary open-angle glaucoma, retinal light damage, and retinopathy of prematurity. These ocular diseases are the cause of irreversible blindness worldwide. Oxidative stress, inflammation and autophagy are implicated in biochemical and morphological changes in these ocular tissues. The development of therapy is a major target for the management care of these ocular diseases.


Binarization of low-quality barcode images captured by mobile phones using local window of adaptive location and size.
It is difficult to directly apply existing binarization approaches to the barcode images captured by mobile device due to their low quality. This paper proposes a novel scheme for the binarization of such images. The barcode and background regions are differentiated by the number of edge pixels in a search window. Unlike existing approaches that center the pixel to be binarized with a window of fixed size, we propose to shift the window center to the nearest edge pixel so that the balance of the number of object and background pixels can be achieved. The window size is adaptive either to the minimum distance to edges or minimum element width in the barcode. The threshold is calculated using the statistics in the window. Our proposed method has demonstrated its capability in handling the nonuniform illumination problem and the size variation of objects. Experimental results conducted on 350 images captured by five mobile phones achieve about 100% of recognition rate in good lighting conditions, and about 95% and 83% in bad lighting conditions. Comparisons made with nine existing binarization methods demonstrate the advancement of our proposed scheme.


Highly oxygenated guaiane-type sesquiterpene lactones from Artemisia sacrorum and their antihepatoma activity.
The ethanol and EtOAc extracts of Artemisia sacrorum exhibited inhibitory effect against HepG2, Huh7, and SK-Hep-1 cell lines with inhibitory ratios of 65.5%, 28.1%, 84.6%, and 93.5%, 82.0%, 89.0% at 200 μg/mL. Twenty-three undescribed guaiane-type sesquiterpene lactones, artemisacrolides A‒W, were isolated from A. sacrorum under the guidance of antihepatoma activity. Their structures were elucidated by spectral data (HRESIMS, IR, UV, 1D and 2D NMR), ECD calculations, and a single-crystal X-ray diffraction. Artemisacrolides A‒U were guaiane-type sesquiterpene lactones possessing α-methylene-γ-lactone and containing acetoxyl groups at C-8, and artemisacrolides V and W represented the first report from the genus Artemisia with a 1,10-rearranged guaiane-type sesquiterpene lactone. Antihepatoma assay suggested that artemisacrolides A‒U demonstrated better inhibitory activity in Huh7 and SK-Hep-1 cells than those of HepG2 cells. Among them, nine compounds exhibited significant inhibitory activity against Huh7 cells with IC50 values of 8.2-14.3 μM, superior or equal to that of sorafenib; seven compounds demonstrated obvious activity against SK-Hep-1 cells with IC50 values of 13.5-19.2 μM, which were equivalent to that of sorafenib. Artemisacrolides B and E were the most active ones in three human hepatoma cell lines with IC50 values of 21.9, 8.2, 16.9 and 22.6, 9.0, 17.3 μM.


Risk of major adverse perinatal outcomes in women with eating disorders.
Low birth weight, prematurity and higher miscarriage rates have previously been reported in women with eating disorders. To determine whether women with a history of eating disorders are at higher risk of major adverse perinatal outcomes. Adjusted birth weight, preterm delivery and miscarriage history were compared in those with a history of eating disorders (anorexia nervosa (n=171), bulimia nervosa (n=199) and both (n=82)) and those with other (n=1166) and no psychiatric disorders (n=10 636) in a longitudinal cohort study. The group with bulimia nervosa had significantly higher rates of past miscarriages (relative risk ratio 2.0, P=0.01) and the group with anorexia nervosa delivered babies of significantly lower birth weight than the general population (P=0.01), which was mainly explained by lower pre-pregnancy body mass index. Preterm delivery rates were comparable across groups. Women with a history of eating disorders are at higher risk of major adverse obstetric outcomes. Antenatal services should be aware of this higher risk.


The development of novel hepatitis B vaccines.
Development of vaccines against viral hepatitis B has proceeded along four main lines. (1) Human plasma-derived vaccines are safe, effective and are now in general use. (2) Subunit polypeptide vaccines formulated in micelles have reached the stage of clinical trials. (3) Recombinant DNA vaccines have been produced in prokaryotic and eukaryotic cells, notably in yeast. The yeast-derived recombinant vaccines have proved safe and effective in extensive clinical trials, eliciting antibodies which in quantity and specificity are equal to those elicited by plasma-derived vaccine. DNA recombinant has also been applied to the development of hybrid and vaccinia virus vaccines which are capable of immunological "priming", and other hybrid virus vaccines are under development. (4) Finally, chemical synthesis has succeeded in producing small peptides which include specific epitopes eliciting antibody responses in experimental animals. Such chemically synthesized preparations offer a prospect of ultimately producing multivalent synthetic vaccines against several viruses, bacteria and protozoa.


Patterns of drug use during a 15 year period: data from a Swedish county, 1988--2002.
To present and interpret drug prescription patterns, related to various groups of the population in a Swedish county, in order to estimate the prevalence of drug use in different age groups. Data on prescriptions, dispensed March-May 1988-2002, were combined with population statistics of Halland, a county in the south of Sweden, and analysed. Number of defined daily doses (DDD) per 100 inhabitants and day and prescriptions per 100 inhabitants and 3 months were used as indicators of drug prevalence. The total drug exposure in the population of Halland nearly doubled during the 15-year period. The most frequently used drugs overall, in 2002, were psycholeptics (N05), analgesics (N02), antibacterials (J01) and sex hormones (G03). Nearly 30% of the women of 15-69 years were exposed to sex hormones. Multiplied drug prevalence among people above 60 was found for antithrombotic drugs (B01), agents acting on the renin-angiotensin system (C09), sex hormones (G03), serum lipid reducing agents (C10), antidepressants (N06) and drugs for peptic ulcer and GORD (A02B). The increase in drug prescribing over the 15 years concerned both symptom-related treatments, like hormone replacement therapy, analgesics, antidepressants and drugs for acid-related disorders, as well as preventive treatments, like antithrombotics, lipid-lowering drugs and antihypertensives. The unit DDD/100 inhabitants and day gives a fairly correct measure of the percentage treated for chronic disorders. However, for short-term treatment courses and especially for drug use in children, number of prescriptions/100 inhabitants and adequate period of time, is easier to interpret.


A metabolomic approach to study major metabolite changes during acclimation to limiting CO2 in Chlamydomonas reinhardtii.
Using a gas chromatography-mass spectrometry-time of flight technique, we determined major metabolite changes during induction of the carbon-concentrating mechanism in the unicellular green alga Chlamydomonas reinhardtii. In total, 128 metabolites with significant differences between high- and low-CO(2)-grown cells were detected, of which 82 were wholly or partially identified, including amino acids, lipids, and carbohydrates. In a 24-h time course experiment, we show that the amino acids serine and phenylalanine increase transiently while aspartate and glutamate decrease after transfer to low CO(2). The biggest differences were typically observed 3 h after transfer to low-CO(2) conditions. Therefore, we made a careful metabolomic examination at the 3-h time point, comparing low-CO(2) treatment to high-CO(2) control. Five metabolites involved in photorespiration, 11 amino acids, and one lipid were increased, while six amino acids and, interestingly, 21 lipids were significantly lower. Our conclusion is that the metabolic pattern during early induction of the carbon-concentrating mechanism fit a model where photorespiration is increasing.


Extinct (129)I in halite from a primitive meteorite: evidence for evaporite formation in the early solar system
Halite crystals from the Zag H3-6 chondrite contain essentially pure (monoisotopic) xenon-129 ((129)Xe) produced in the early history of the solar system by the decay of short-lived iodine-129 ((129)I) (half-life = 15.7 million years). Correlated release of (129)Xe and (128)Xe, produced artificially from (127)I by neutron irradiation, corresponds to an initial ((129)I/(127)I) ratio of (1.35 +/- 0.05) x 10(-4), close to the most primitive early solar system value. If the (129)Xe was produced by in situ decay, then the halite formed from an aqueous fluid within 2 million years of the oldest known solar system minerals.


Ibutilide Effectiveness and Safety in the Cardioversion of Atrial Fibrillation and Flutter in the Community Emergency Department.
Little is known about the use of ibutilide for cardioversion in atrial fibrillation and flutter outside of clinical trials. We seek to describe patient characteristics, ibutilide administration patterns, cardioversion rates, and adverse outcomes in the community emergency department (ED) setting. We also evaluate potential predictors of cardioversion success. Using a retrospective cohort of adults who received ibutilide in 21 community EDs between January 2009 and June 2015, we gathered demographic and clinical variables from electronic health records and structured manual chart review. We calculated rates of cardioversion and frequency of ventricular tachycardia within 4 hours and estimated adjusted odds ratios (aOR) in a multivariate regression model for potential predictors of cardioversion. Among 361 patients, the median age was 61 years (interquartile range 53 to 71 years) and most had recent-onset atrial fibrillation and flutter (98.1%). Five percent of the cohort had a history of heart failure. The initial QTc interval was prolonged (>480 ms) in 29.4% of patients, and 3.1% were hypokalemic (<3.5 mEq/L). The mean ibutilide dose was 1.5 mg (SD 0.5 mg) and the rate of ibutilide-related cardioversion within 4 hours was 54.8% (95% confidence interval [CI] 49.6% to 60.1%), 50.5% for atrial fibrillation and 75.0% for atrial flutter. Two patients experienced ventricular tachycardia (0.6%), both during their second ibutilide infusion. Age (in decades) (aOR 1.3; 95% CI 1.1 to 1.5), atrial flutter (versus atrial fibrillation) (aOR 2.7; 95% CI 1.4 to 5.1), and no history of atrial fibrillation and flutter (aOR 2.0; 95% CI 1.2 to 3.1) were associated with cardioversion. The effectiveness and safety of ibutilide in this community ED setting were consistent with clinical trial results despite less stringent patient selection criteria.


[Complete neuroleptic withdrawal in patients with schizophrenia with intense symptoms and resistant to treatment].
We report 2 studies evaluating the effects of a complete and progressive neuroleptic withdrawal on the symptomatology of 2 groups of 10 young chronically ill schizophrenic inpatients. In a preliminary open study, we compare the psychiatric symptomatology during a 4-week-period before the beginning of withdrawal and during a similar period following the end of withdrawal. We observe the significative improvement of the blunted affect, the deterioration of an aspecific psychiatric symptomatology (including irritability, excitement, hostility) and the non-modification of the specific schizophrenic symptomatology (in its 3 main components: positive signs, negative signs, disorganization). The second study (a double blind one) reports the effects of a complete neuroleptic withdrawal maintained during a longer period (8 weeks): 1. The improvement of the blunted affect is transiently observed during the 2 first weeks after the end of withdrawal but not after this time. 2. The deterioration of the aspecific psychiatric symptomatology by opposition to the non-significative modification of the specific schizophrenic symptomatology is a fact confirmed by this second study. Even if we can not exclude that the influence of neuroleptic withdrawal on the specific psychotic symptoms may require a longer time frame than 8 weeks to detect, this dichotomic evolution raises the problem of the specific action of neuroleptics in this particular category of patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Preferential role of calcium in the regulation of prolactin gene transcription by thyrotropin-releasing hormone in GH3 pituitary cells.
TRH induces two separate events in pituitary PRL cells. It increases the release of stored PRL and enhances the rate of PRL gene transcription, which results in an increased steady state concentration of PRL messenger RNA (mRNA) and a concomitant augmentation of PRL production. The mechanisms underlying the release process involve the activation of phosphatidylinositol turnover which generates inositol 1,4,5-trisphosphate and 1,2-diacylglycerol. In order to determine whether these intracellular messengers also mediate the stimulation of PRL gene expression by TRH, we have correlated the level of receptor occupancy with the rate of gene transcription and investigated the action of drugs which increase cytosolic calcium or activate protein kinase C. We have determined that sustained stimulation of transcription requires the persistent occupancy of a limited number of TRH receptor sites and that the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), calcium ionophores (A23187, ionomycin), and the calcium channel agonist BAY K 8644 enhance PRL gene transcription. However, TPA is less potent and ionomycin requires a low concentration of TPA to fully mimic TRH action, whereas BAY K 8644 alone displays the same potency as TRH. The effects of BAY K 8644 and TRH are not additive and thus suggest that the influx of calcium plays a predominant role in the regulation of PRL gene transcription by TRH.


[Relationship between beta-lactamase activity of clinical strains of Pseudomonas pyocyanea and their resistance to penicillins].
The study of 50 clinical strains of Ps. aeruginosa revealed their high resistance to benzylpenicillin, ampicillin, oxacillin, streptomycin and kanamycin with a tendency to polyresistance. The same strains were highly sensitive to gentamycin and polymyxin. The strains produced constitutive beta-lactamase in small amounts. The enzyme synthesis in Ps. aeruginosa was induced by benzylpenicillin. Still high concentrations of it were required for the induction. The maximum induction of beta-lactamase synthesis was observed by the 6th hour of the inductor addition. The maximum induction level was observed 4 hours after addition of benzylpenicillin. The level of induction of beta-lactamase synthesis in the strains ranged within 5-93. A significant part of the enzyme was liberated from the cells during induction. Interaction between the induction level of beta-lactamase synthesis and resistance of Ps. aeruginosa to penicillins was found.


Knowledge and Acceptability about Adult Pertussis Immunization in Korean Women of Childbearing Age.
The adult tetanus, reduced diphtheria, and acellular pertussis (Tdap) vaccine has been introduced in order to provide individual protection and reduce the risk of transmitting pertussis to infants. We assessed the knowledge and acceptability of the Tdap vaccine around pregnancy. This study was a cross-sectional survey of women of childbearing age (20-45 years) who visited obstetrics and gynecologic units of primary, secondary, or tertiary hospitals. They were asked to fill in a questionnaire assessing their knowledge, attitudes, and acceptability of Tdap. The questionnaire was completed by 308 women; 293 (95.1%) had not received information from doctors about Tdap, and 250 (81.2%) did not know about the need for vaccination. A significantly important factor related to subjects' intention to be vaccinated, identified by stepwise multiple logistic regression, was the knowledge (OR 13.5, CI 3.92-46.33) that adult Tdap is effective in preventing pertussis for infants aged 0-6 months. Additionally, 276 (89.6%) considered the recommendation of obstetric doctors as the most influencing factor about Tdap vaccination. In Korea, most women of childbearing age seem to be neither recommended nor adequately informed about the vaccination, although our population was not a nationwide representative sample. Information given by healthcare workers may be critical for improving awareness and preventing pertussis.


Evaluation of point of care serum cryptococcal antigen by lateral flow immunoassay for diagnosis of cryptococcosis and cryptococcal meningitis in HIV-positive patients.
Cryptococcal meningitis (CM) is the initial acquired immunodeficiency syndrome (AIDS) defining illness in 2% of patients with CD4 levels <100/μL and a leading cause of mortality in AIDS in the developing world. It is the most common opportunistic infection of the central nervous system in AIDS in various Indian studies. Detection of serum cryptococcal antigen (SCRAG) is the most widely used diagnostic method for cryptococcosis. The presence of cerebrospinal fluid cryptococcal antigen (CSF CRAG) is diagnostic of CM. CRAG can be determined by latex agglutination (LAT), enzyme-linked immunosorbent assay and now, by lateral flow (LFA)immunoassay. LFA is a point of care test that rapidly detects CRAG. This study compares LAT and LFA for the detection of serum CRAG and diagnosing CM. Two hundred and ten patients of HIV/AIDS were submitted to SCRAG LFA by dipstick. A sample was also sent to laboratory for SCRAG by LAT. CSF examination was done for those who were positive for SCRAG LFA and those who had symptoms suggestive of meningitis. SCRAG by LFA was compared with SCRAG by LAT, CSF CRAG by LAT and LFA, CSF cryptococcal culture and CSF India ink examination for Cryptococcus. Fifteen patients were found positive for SCRAG by LFA dipstick. All of them were also positive for SCRAG by LAT. Twelve of them had C. D4 count below below 100 cells/mm3. CSF CRAG was positive in all 12 SCRAG positive who were submitted to CSF examination. We found that serum detection of CRAG by LFA dipstick is as sensitive as CRAG detection in serum by LAT and CSF CRAG detection by LFA and LAT. It is thus a rapid test for diagnosing CM in HIV patients with low CD4 counts.


Retinopathy of prematurity: screening and treatment in Costa Rica.
To determine the recent demographic data, risk factors and results of retinopathy of prematurity (ROP) screening and treatment in Costa Rica. The medical records of all preterm infants meeting ROP screening criteria (≤34 weeks' gestational age (GA) or birth weight (BW) ≤1750g, and those determined at risk by neonatologists) in the national healthcare system, Costa Rica, January 2010-December 2014, were retrospectively reviewed. The numbers and percentages of infants with ROP, risk factors, percentage of patients treated and treatment outcomes were determined. Comparison is made with screening criteria and literature reports of ROP incidence in other countries. The study population included 3018 preterm infants. Overall, 585 patients (585/3018, 19.4%) were found to have ROP. Of these, 15.4% (90 patients) required laser treatment, and 53% of those requiring treatment had BW <1000g. Five babies requiring treatment were ≥32 weeks' GA but with BW ≤1750g. Aggressive posterior disease was found in nine patients, and two infants of those screened (2/3018, 0.07%) suffered severe visual impairment during the 5-year study period. We provide comprehensive data of ROP care in Costa Rica allowing assessment and comparison of screening criteria and protocol.


Mean-deviation analysis in the theory of choice.
Mean-deviation analysis, along with the existing theories of coherent risk measures and dual utility, is examined in the context of the theory of choice under uncertainty, which studies rational preference relations for random outcomes based on different sets of axioms such as transitivity, monotonicity, continuity, etc. An axiomatic foundation of the theory of coherent risk measures is obtained as a relaxation of the axioms of the dual utility theory, and a further relaxation of the axioms are shown to lead to the mean-deviation analysis. Paradoxes arising from the sets of axioms corresponding to these theories and their possible resolutions are discussed, and application of the mean-deviation analysis to optimal risk sharing and portfolio selection in the context of rational choice is considered.


Accumulation of pigment granules around nuclei in early embryos of Anura (Amphibia).
A dark spot was found to appear in each blastomere of the vegetal surface of blastulae of Rana rugosa, Hyla arborea. The spot divided into two before division of the blastomere, so that one new spot was allotted to each daughter cell. These dark spots were formed at early blastula stage, and persisted until the end of yolk plug stage. Cytological observations showed that each dark spot corresponds to a mass of accumulated pigment granules around the nucleus of a blastomere. The accumulation increases with development during the cleavage period more rapidly in blastomeres of the vegetal hemisphere than in those of the animal hemisphere. This accumulation of pigment granules around nuclei during development indicates that the granules are transported toward the nuclei during the cleavage period, suggesting some sort of directional flow of cytoplasm in blastomeres of early amphibian embryos.


Structural determination of 5'-OH alpha-ribofuranoside modified cobalamins via 13C and DEPT NMR.
A protocol for the rapid NMR characterization of cobalamin (vitamin B(12)) analogues with 5'-hydroxy-alpha-ribofuranoside modification is reported. The structure of cyanocobalamin in DMSO-d(6) has been assigned using COSY, NOESY, HSQC, and HMBC NMR methods. The robust precision of (13)C NMR assignments in DMSO-d(6) allows for the rapid structural determination of 5'-hydroxy-alpha-ribofuranosyl cyanocobalamin derivatives with solely 1-D (13)C and DEPT NMR spectra and only 10 mg of derivatized cobalamin. Using this method, the (13)C NMR resonances of four cobalamin analogues were determined with the most significant variance of (13)C chemical shifts occurring in the alpha-ribofuranoside ring. In DMSO-d(6), cobalamin concentrations greater than 30 mM can be achieved for an improved signal-to-noise ratio.


Royal College debates whether MDs should promote moderate consumption of alcohol.
Emerging news about the potential beneficial effects of moderate alcohol consumption raises some interesting challenges for physicians, who often come face to face with problems created by alcohol. Physicians on the affirmative side won a debate on the pros and cons of moderate drinking that was held during this fall's Royal College meeting. Pam Harrison explains how they did it.


Down-regulation of 3-phosphoinositide-dependent protein kinase-1 levels inhibits migration and experimental metastasis of human breast cancer cells.
High expression of 3-phosphoinositide-dependent protein kinase-1 (PDK1) has been detected in various invasive cancers. In the current study, we investigated its role in cancer cell migration and experimental metastasis. Down-regulation of PDK1 expression by small interference RNA markedly inhibited spontaneous migration and epidermal growth factor (EGF)-induced chemotaxis of human breast cancer cells. The defects were rescued by expressing wild-type PDK1. PDK1-depleted cells showed impaired EGF-induced actin polymerization and adhesion, probably due to a decrease in phosphorylation of LIM kinase/cofilin and integrin beta1. Confocal microscopy revealed that EGF induced cotranslocation of PDK1 with Akt and protein kinase Czeta (PKCzeta), regulators of LIM kinase, and integrin beta1. Furthermore, PDK1 depletion dampened EGF-induced phosphorylation and translocation of Akt and PKCzeta, suggesting that Akt and PKCzeta functioned downstream of PDK1 in the chemotactic signaling pathway. In severe combined immunodeficiency mice, PDK1-depleted human breast cancer cells formed more slowly growing tumors and were defective in extravasation to mouse lungs after i.v. injection. Our results indicate that PDK1 plays an important role in regulating the malignant behavior of breast cancer cells, including their motility, through activation of Akt and PKCzeta. Thus, PDK1, which increases its expression in cancer cells, can be used as a target for the development of novel therapies.


[Combined treatment of type 2 diabetes].
Type 2 diabetes at present represents a challenge to public health throughout the world. Its incidence is increasing in a considerable way and, therefore, cardiovascular morbi-mortality associated with macro or micro-angiopathy. The prevention of vascular complications in these patients must include control of all the cardiovascular risk factors as well as of glycemia. In recent years it has been shown that the control of the latter has to be much more strict and that in many cases monotherapy does not achieve the established aims. At the same time, new oral anti-diabetic medicines have recently appeared with different mechanisms of action. The possible combinations of treatment with different oral anti-diabetics, or else with oral anti-diabetics with insulin, are very numerous and have shown their effectiveness in reducing glycemia and the glycosylated haemoglobin. This is why the combined treatment is at present an alternative to insulinisation in monotherapy for patients with type 2 diabetes and bad control in monotherapy. In this paper the available scientific evidence on each one of the combinations and their possible indications is reviewed. Selection of the type of association necessary will depend on the individual aims of control, on the physiopathological mechanism presumably involved in each case, on the efficacy, cost and secondary effects of each medicine, as well as on the characteristics of each patient.