Effects of repeated use and resterilization on structural and functional integrity of microwave ablation antennas.
OBJECTIVE To determine effects of repeated use and resterilization on structural and functional integrity of microwave ablation (MWA) antennas. SAMPLE 17 cooled-shaft MWA antennas (3 groups of 5 antennas/group and 2 control antennas). PROCEDURES 1, 2, and 3 ablations in the livers of bovine cadavers were performed at the maximum recommended settings. Antennas were cleaned and sterilized in hydrogen peroxide plasma, and the process was repeated (reprocessing cycle; n = 6). Control antennas were only sterilized (6 times). Aerobic and anaerobic bacterial cultures were performed, and antennas were microscopically assessed for damage. RESULTS 6 cycles were completed. Thirteen of 15 MWA antennas remained functional for up to 4 cycles, 10 were functional after 5 cycles, and only 7 were functional after 6 cycles. Progressive tearing of the silicone coating of the antennas was observed, with a negative effect of the number of cycles for silicone tearing. Size of the ablation zone decreased mildly over time after cycles 5 and 6; however, this was not considered clinically relevant. No significant changes in the shape of ablation zones were detected. All cultures yielded negative results, except for an isolated case, which was considered a contaminant. CONCLUSIONS AND CLINICAL RELEVANCE Structural and functional integrity of the microwave antennas remained acceptable during repeated use and reprocessing for up to 4 cycles. However, there was a decrease in functional integrity at cycles 5 and 6. We suggest that these microwave antennas be subjected to > 3 reprocessing cycles. Antennas should be carefully examined before reuse.


Fos induction in the Japanese quail brain after expression of appetitive and consummatory aspects of male sexual behavior.
We investigated the expression of Fos, the protein product of the immediate early gene c-fos in the brain of male Japanese quail after they engaged in either appetitive or consummatory sexual behavior (i. e., copulation). For 1 h, castrated males treated with testosterone were either allowed to copulate with a female or to exhibit a learned social proximity response indicative of appetitive sexual behavior. Control birds were either left in their home cage or placed in the experimental chamber but did not exhibit the appetitive sexual behavior because they had never learned it. Fos expression was studied with an immunocytochemical procedure in two sets of adjacent sections through the entire forebrain. These sections were immunolabelled with 2 different antibodies raised against a synthetic fragment corresponding to the 21 carboxy-terminal residues of the chicken Fos sequence. Contrary to the results of a previous study in which gonadally intact birds were used, Fos induction was observed neither in the medial preoptic nucleus nor in the nucleus intercollicularis in birds that had interacted for 1 h with a female. This may be related to a lower frequency of copulation in the testosterone-implanted birds than in intact birds, or to differences in the time the brains were collected after the birds engaged in sexual behavior between the two studies (60 min in this study, 120 min in the previous study). The performance of copulation and/or appetitive sexual behavior increased the number of Fos-immunoreactive cells in the ventral hyperstriatum, medial archistriatum, and nucleus striae terminalis. These increases were observed using both antibodies, although each antibody produced minor differences in the number of Fos-immunoreactive cells observed. Using one of the antibodies, but not the other, increases in Fos immunoreactivity were also observed in the nucleus accumbens and hyperstriatum after either copulation or appetitive sexual behavior. These differences illustrate how minor technical variations in the Fos immunocytochemical procedure influence the results obtained. These differences also show that Fos induction in a number of brain regions is observed after performance of consummatory (copulation) as well as appetitive (looking at the female) sexual behavior. This induction is, therefore, not related solely to the control of copulatory acts but, presumably, also to the processing in a variety of telencephalic association areas of stimuli originating from the female. The observation that increased Fos immunoreactivity is present in birds that had learned the response indicative of appetitive sexual behavior, and not in those that had not learned the behavior, further indicates that it is not simply the sight of the female that results in this Fos induction, but the analysis of the relevant stimuli in a sexually explicit context. Conditioned neural activity resulting from a learned association between the stimulus female and the performance of copulatory behavior may also explain some aspects of the brain activation observed in birds viewing, but not allowed to interact with, the female.


Physical activity patterns and obesity status among 10- to 12-year-old adolescents living in Athens, Greece.
Childhood obesity has become a modern epidemic with escalating rates. The aim of our study was to identify physical activity patterns among Greek schoolchildren and to examine their relationship with obesity. 700 adolescents age 10 to 12 years were evaluated through a standardized questionnaire. Several demographic, socioeconomic, and physical activity characteristics were recorded. Physical activity was assessed and adolescents were characterized as active and nonactive. Body height and weight were measured and body mass index was calculated in order to classify subjects as overweight or obese (IOTF classification). Multiple logistic regression and multivariate techniques (principal components analysis) were performed. Eight physical activity patterns were identified, including increased physical activity in weekdays and weekends, sports physical activity, vigorous, moderate, and low physical activity. Increased physical activity on weekends and vigorous physical activity in boys were negatively associated with being overweight or obese (OR: 0.65; 95% CI: 0.48-0.90 and OR: 0.66; 95% CI: 0.49-0.88, correspondingly) and moderate physical activity was marginally positively associated in girls (OR: 1.28; 95% CI: 0.97-1.69), after adjusting for several confounders. Our findings demonstrate the important role of vigorous physical activity in the maintenance of normal weight of adolescents.


Synthesis and Properties of Quinoidal Fluorenofluorenes.
The synthesis and optoelectronic properties of 24 π-electron, formally antiaromatic fluoreno[3,2-b]fluorene and fluoreno[4,3-c]fluorene (FF), are presented. The solid-state structure of [4,3-c]FF along with computationally analogous molecules shows that the outer rings are aromatic while the central four rings possess a bond-localized 2,6-naphthoquinodimethane motif. The antiaromaticity and biradical character of the FFs is assessed computationally, the results of which indicate the dominance of the closed-shell ground state for these molecules.


Theoretical investigation of substituted anthraquinone dyes.
We have investigated with computational chemistry techniques the visible spectra of substituted anthraquinones. A wide panel of theoretical methods has been used, with various basis sets and density functional theory (DFT) functionals, in order to assess a level of theory that would lead to converged excitation energies. It turns out that the hybrid Becke-Lee-Yang-Parr and Perdew-Burke-Erzenrhof functionals with the 6-31G (d,p) atomic basis set provide reliable lambda(max) when the solvent effects are included in the model. Combining the results of both DFT schemes allows the prediction of lambda(max) with a standard deviation limited to 13 nm.


Chronic Peptide Therapy With B-Type Natriuretic Peptide in Patients With Pre-Clinical Diastolic Dysfunction (Stage B Heart Failure).
This study determined whether there is development of tachyphylaxis to enhancement of cardiorenal response to acute volume loading (AVL) with B-type natriuretic peptide (BNP) after 12-week, twice-daily subcutaneous BNP administration in patients with preclinical diastolic dysfunction (PDD). PDD is characterized by normal systolic function and moderate or severe diastolic dysfunction but no symptoms of heart failure (HF). Impairment in cardiorenal endocrine response to stress by AVL exists in PDD and is corrected by acute administration of subcutaneous BNP. A double-blinded, placebo-controlled proof-of-concept study was conducted to compare 12 weeks of twice daily subcutaneous BNP, 10 μg/kg (n = 24), versus placebo (n = 12) in PDD. Subjects underwent 2 study visits, at baseline and after 12 weeks. At each study visit, echocardiography, renal, and neurohumoral assessments were performed before and after intravascular AVL. Among those with PDD, there was a statistically significant improvement in diastolic function after 12 weeks of BNP, as measured by a decrease in the Doppler E/e' ratio (where E is early mitral inflow velocity and e' is mitral annulus early diastolic motion) (p = 0.004) and improvement of diastolic dysfunction grade (p = 0.008). After 12 weeks, there was statistically significantly greater sodium excretion, urine flow, and urinary cyclic guanosine monophosphate excretion to AVL (all p < 0.001), as well as a trend toward greater glomerular filtration rate (p = 0.050) in the BNP group as compared to the placebo group. In subjects with PDD, chronic BNP administration resulted in sustained improvement in diastolic function without development of tachyphylaxis to the enhancement of cardiorenal response to volume expansion with BNP. (Human Brain Natriuretic Peptide [BNP] [or Nesiritide] to Help Heart, Kidney and Humoral Function; NCT00405548).


Children with vesical rhabdomyosarcoma (RMS) treated by partial cystectomy with neoadjuvant or adjuvant chemotherapy, with or without radiotherapy. A report from the Intergroup Rhabdomyosarcoma Study (IRS) Committee.
Children with bladder rhabdomyosarcoma (RMS) are currently treated with primary chemotherapy and usually with local irradiation. More than 30% of this group ultimately require total cystectomy. The purpose of this study was to review the results of the use of partial as opposed to total cystectomy in 40 patients with bladder RMS during Intergroup Rhabdomyosarcoma Study (IRS)-I, -II, and -III (1972-1989). A total of 171 children with primary RMS of the bladder were enrolled in the IRS during this interval. Partial cystectomy was performed in 40, in 33 before any other therapy and in seven after 10-57 weeks of chemotherapy (primarily vincristine, actinomycin-D, and cyclophosphamide, i.e., VAC) +/- radiotherapy. Surviving patients have been observed for 4-22 years. Thirty-one of 40 patients (78.5%) have been disease free for 2-16 years. Survival among all other IRS cases with bladder RMS during the same interval was 79.5%. Of the 31 surviving patients, one required secondary total cystectomy and two required bladder augmentation procedures for benign bladder contracture. Three quarters of the total group of living patients who have undergone partial cystectomy are without bladder-related symptoms or demonstrable lower urinary tract disease. The remaining patients have a history of functional bladder problems related to contracture or incontinence. Partial cystectomy is an alternative to total cystectomy for bladder RMS when the tumor site makes it anatomically feasible. In such patients, it should be considered before total cystectomy and in patients with persistent areas of questionable residual tumor after otherwise apparently successful regimens of primary chemotherapy +/- radiotherapy.


Bimodal dynamics of granular organelles in primary renin-expressing cells revealed using TIRF microscopy.
Renin is the initiator and rate-limiting factor in the renin-angiotensin blood pressure regulation system. Although renin is not exclusively produced in the kidney, in nonmurine species the synthesis and secretion of the active circulatory enzyme is confined almost exclusively to the dense core granules of juxtaglomerular (JG) cells, where prorenin is processed and stored for release via a regulated pathway. Despite its importance, the structural organization and regulation of granules within these cells is not well understood, in part due to the difficulty in culturing primary JG cells in vitro and the lack of appropriate cell lines. We have streamlined the isolation and culture of primary renin-expressing cells suitable for high-speed, high-resolution live imaging using a Percoll gradient-based procedure to purify cells from RenGFP+ transgenic mice. Fibronectin-coated glass coverslips proved optimal for the adhesion of renin-expressing cells and facilitated live cell imaging at the plasma membrane of primary renin cells using total internal reflection fluorescence microscopy (TIRFM). To obtain quantitative data on intracellular function, we stained mixed granule and lysosome populations with Lysotracker Red and stimulated cells using 100 nM isoproterenol. Analysis of membrane-proximal acidic granular organelle dynamics and behavior within renin-expressing cells revealed the existence of two populations of granular organelles with distinct functional responses following isoproterenol stimulation. The application of high-resolution techniques for imaging JG and other specialized kidney cells provides new opportunities for investigating renal cell biology.


Self-assembled hydrogel fibers for sensing the multi-compartment intracellular milieu.
Targeted delivery of drugs and sensors into cells is an attractive technology with both medical and scientific applications. Existing delivery vehicles are generally limited by the complexity of their design, dependence on active transport, and inability to function within cellular compartments. Here, we developed self-assembled nanofibrous hydrogel fibers using a biologically inert, low-molecular-weight amphiphile. Self-assembled nanofibrous hydrogels offer unique physical/mechanical properties and can easily be loaded with a diverse range of payloads. Unlike commercially available E. coli membrane particles covalently bound to the pH reporting dye pHrodo, pHrodo encapsulated in self-assembled hydrogel-fibers internalizes into macrophages at both physiologic (37°C) and sub-physiologic (4°C) temperatures through an energy-independent, passive process. Unlike dye alone or pHrodo complexed to E. coli, pHrodo-SAFs report pH in both the cytoplasm and phagosomes, as well the nucleus. This new class of materials should be useful for next-generation sensing of the intracellular milieu.


Shorter and proximal Timeline Followback windows are representative of longer posttreatment functioning.
Very little research has been conducted on what time window provides a representative picture of daily drinking. With respect to pretreatment drinking, one study that used the Timeline Followback (TLFB) with problem drinkers found that a 3-month window is generally representative of annual pretreatment drinking. The objective of the present study was to determine the shortest representative time window for reports of annual posttreatment drinking. A second objective was to determine which of two time windows, 90 days from the end of treatment or 90 days prior to the end of follow-up, was the most representative proxy for annual posttreatment drinking. TLFB reports from 467 problem drinkers who participated in a randomized controlled trial of a mail-based intervention were used in the present analysis. The results show that a 3-month posttreatment window (i.e., first 90 days after the intervention) is sufficiently representative (r = .94) of annual posttreatment drinking for problem drinkers (i.e., less severely dependent alcohol abusers). In addition, although there were no clinically significant differences in drinking behavior between the two 90-day posttreatment windows, the use of proximal windows (i.e., closer to the end of treatment) would minimize participant attrition. In addition, a 3-month versus 12-month TLFB follow-up time frame resulted in a much higher percentage of participants completing the full TLFB (89% vs. 71%). Further research is needed to determine if these findings will generalize to more severely dependent alcohol abusers.


Case-oriented computer-based-training in radiology: concept, implementation and evaluation.
Providing high-quality clinical cases is important for teaching radiology. We developed, implemented and evaluated a program for a university hospital to support this task. The system was built with Intranet technology and connected to the Picture Archiving and Communications System (PACS). It contains cases for every user group from students to attendants and is structured according to the ACR-code (American College of Radiology) 2. Each department member was given an individual account, could gather his teaching cases and put the completed cases into the common database. During 18 months 583 cases containing 4136 images involving all radiological techniques were compiled and 350 cases put into the common case repository. Workflow integration as well as individual interest influenced the personal efforts to participate but an increasing number of cases and minor modifications of the program improved user acceptance continuously. 101 students went through an evaluation which showed a high level of acceptance and a special interest in elaborate documentation. Electronic access to reference cases for all department members anytime anywhere is feasible. Critical success factors are workflow integration, reliability, efficient retrieval strategies and incentives for case authoring.


TcG2/TcG4 DNA Vaccine Induces Th1 Immunity Against Acute Trypanosoma cruzi Infection: Adjuvant and Antigenic Effects of Heterologous T. rangeli Booster Immunization.
Background: Chagas cardiomyopathy is caused by Trypanosoma cruzi (Tc). Two antigenic candidates, TcG2 and TcG4, are recognized by antibodies in naturally infected dogs and humans; and these vaccine candidates provided protection from Tc infection in mice and dogs. Trypanosoma rangeli (Tr) is non-pathogenic to mammals and shown to elicit cross-reactive anti-Tc antibodies. In this study, we investigated if fixed Tr (fTr) can further enhance the efficacy of the TcG2/TcG4 DNA vaccine. Methods and Results: C57BL/6 mice were immunized with TcG2/TcG4 DNA vaccine and fTr (delivered as an adjuvant or in prime-boost approach), and challenged with Tc. Serology studies showed that fTr (±quil-A) elicited Tc- and Tr-reactive IgGs that otherwise were not stimulated by TcG2/TcG4 vaccine only, and quil-A had suppressive effects on fTr-induced IgGs. After challenge infection, TcG2/TcG4-vaccinated mice exhibited potent expansion of antigen- and Tc-specific IgGs that were not boosted by fTr±quil-A. Flow cytometry analysis showed that TcG2/TcG4-induced dendritic cells (DC) and macrophages (Mφ) responded to challenge infection by expression of markers of antigen uptake, processing, and presentation, and production of pro-inflammatory cytokines. TcG2/TcG4-induced CD4+T cells acquired Th1 phenotype and expressed markers that orchestrate adaptive immunity. A fraction of vaccine-induced CD4+T cells exhibited iTreg phenotype responsible for aversion of self-injurious immune responses. Further, TcG2/TcG4-vaccinated mice exhibited potent expansion of poly-functional CD8+T cells with TNF-α/IFN-γ production and cytolytic phenotype post-infection. Subsequently, tissue parasites and pathology were hardly detectable in TcG2/TcG4-vaccinated/infected mice. Inclusion of fTr±quil-A had no clear additive effects in improving the Tc-specific adaptive immunity and parasite control than was noted in mice vaccinated with TcG2/TcG4 alone. Non-vaccinated mice lacked sufficient activation of Th1 CD4+/CD8+T cells, and exhibited >10-fold higher levels of tissue parasite burden than was noted in vaccinated/infected mice. Conclusion:TcG2/TcG4 vaccine elicits highly effective immunity, and inclusion of fTr is not required to improve the efficacy of DNA vaccine against acute Tc infection in mice.


X-Linked adrenoleukodystrophy: overview and prognosis as a function of age and brain magnetic resonance imaging abnormality. A study involving 372 patients.
The phenotypic expression of X-linked adrenoleukodystrophy (X-ALD) ranges from the rapidly progressive childhood cerebral form to the milder adrenomyeloneuropathy (AMN) in adults. It is not possible to predict phenotype by mutation analysis or biochemical assays. This study reports on 372 patients ranging in age from less than 3 years to adulthood, who have been followed at the Kennedy Krieger Institute. With the aim of determining whether a method could be developed to predict clinical course by analysis of data available at time of first contact, the patients were subdivided into 18 subgroups on the basis of age and the extent of brain magnetic resonance (MRI) abnormality utilizing the MRI scoring system devised by Loes et al. Scores to grade degree of neurologic and neuropsychologic impairment were also developed. There was strong correlation between MRI and the neurology and neuropsychology scores at baseline. Information based exclusively on age and MRI score at time of first contact was highly predictive of future clinical course and should aid the evaluation of the effects of bone marrow transplantation and the selection of patients for this procedure, as well as the evaluation of other therapies that may be developed in the future.


Cesium-containing triple cation perovskite solar cells: improved stability, reproducibility and high efficiency.
Today's best perovskite solar cells use a mixture of formamidinium and methylammonium as the monovalent cations. With the addition of inorganic cesium, the resulting triple cation perovskite compositions are thermally more stable, contain less phase impurities and are less sensitive to processing conditions. This enables more reproducible device performances to reach a stabilized power output of 21.1% and ∼18% after 250 hours under operational conditions. These properties are key for the industrialization of perovskite photovoltaics.


Role of HIV infection duration and CD4 cell level at initiation of combination anti-retroviral therapy on risk of failure.
The development of HIV drug resistance and subsequent virological failure are often cited as potential disadvantages of early cART initiation. However, their long-term probability is not known, and neither is the role of duration of infection at the time of initiation. Patients enrolled in the UK Register of HIV seroconverters were followed-up from cART initiation to last HIV-RNA measurement. Through survival analysis we examined predictors of virologic failure (2HIV-RNA ≥400 c/l while on cART) including CD4 count and HIV duration at initiation. We also estimated the cumulative probabilities of failure and drug resistance (from the available HIV nucleotide sequences) for early initiators (cART within 12 months of seroconversion). Of 1075 starting cART at a median (IQR) CD4 count 272 (190,370) cells/mm(3) and HIV duration 3 (1,6) years, virological failure occurred in 163 (15%). Higher CD4 count at initiation, but not HIV infection duration at cART initiation, was independently associated with lower risk of failure (p=0.033 and 0.592 respectively). Among 230 patients initiating cART early, 97 (42%) discontinued it after a median of 7 months; cumulative probabilities of resistance and failure by 8 years were 7% (95% CI 4,11) and 19% (13,25), respectively. Although the rate of discontinuation of early cART in our cohort was high, the long-term rate of virological failure was low. Our data do not support early cART initiation being associated with increased risk of failure and drug resistance.


Aromatic stabilization of functionalized corannulene cations.
The first comprehensive theoretical investigation of aromaticity in functionalized corannulene cations of general formula [CH3-C20H10](+) was accomplished. The experimentally known system [CH3-hub-C20H10](+) was augmented by two other possible isomers, namely, rim- and spoke-ones. Changes in aromaticity, when going from neutral corannulene to its functionalized cations, were monitored with the help of descriptors of different nature such as structure-based HOMA, topological PDI and FLU, and magnetic NICS. A highly efficient tool for analysis and visualization of delocalization and conjugation named ACID was also utilized. In the final step, a complete set of (1)H and (13)C chemical shifts was calculated and compared with the available experimental data. Conservation of aromaticity of 6-membered rings along with vanishing anti-aromatic character of central 5-membered rings was found to be the main reason for the exceptional stability of the hub-isomer. At the same time, functionalization of the corannulene moiety at the rim- or spoke-site resulted in dramatic elimination of aromaticity of 6-membered rings, whereas anti-aromatic character of the central ring remained. Altogether, it led to much lower stability of these isomers in comparison with that of the hub-one.


A Conserved Role for the NAM/miR164 Developmental Module Reveals a Common Mechanism Underlying Carpel Margin Fusion in Monocarpous and Syncarpous Eurosids.
The majority of angiosperms are syncarpous- their gynoecium is composed of two or more fused carpels. In Arabidopsis thaliana, this fusion is regulated through the balance of expression between CUP SHAPED COTYLEDON (CUC) genes, which are orthologs of the Petunia hybrida transcription factor NO APICAL MERISTEM (NAM), and their post-transcriptional regulator miR164. Accordingly, the expression of a miR164-insensitive form of A. thaliana CUC2 causes a radical breakdown of carpel fusion. Here, we investigate the role of the NAM/miR164 genetic module in carpel closure in monocarpous plants. We show that the disruption of this module in monocarpous flowers of A. thaliana aux1-22 mutants causes a failure of carpel closure, similar to the failure of carpel fusion observed in the wild-type genetic background. This observation suggested that closely related mechanisms may bring about carpel closure and carpel fusion, at least in A. thaliana. We therefore tested whether these mechanisms were conserved in a eurosid species that is monocarpous in its wild-type form. We observed that expression of MtNAM, the NAM ortholog in the monocarpous eurosid Medicago truncatula, decreases during carpel margin fusion, suggesting a role for the NAM/miR164 module in this process. We transformed M. truncatula with a miR164-resistant form of MtNAM and observed, among other phenotypes, incomplete carpel closure in the resulting transformants. These data confirm the underlying mechanistic similarity between carpel closure and carpel fusion which we observed in A. thaliana. Our observations suggest that the role of the NAM/miR164 module in the fusion of carpel margins has been conserved at least since the most recent common ancestor of the eurosid clade, and open the possibility that a similar mechanism may have been responsible for carpel closure at much earlier stages of angiosperm evolution. We combine our results with studies of early diverging angiosperms to speculate on the role of the NAM/miR164 module in the origin and further evolution of the angiosperm carpel.


Hyaluronan synthase-3 is upregulated in metastatic colon carcinoma cells and manipulation of expression alters matrix retention and cellular growth.
HA is a glycosaminoglycan that is synthesized on the inner surface of the plasma membrane and secreted into the pericellular matrix. HA and its biosynthetic enzymes (HAS1, HAS2 and HAS3) are thought to participate in tumor growth and cancer progression. In our study, colon carcinoma cells isolated from a lymph node metastasis (SW620) produced more pericellular HA and expressed higher levels of HAS3 mRNA compared to cells isolated from a primary colon carcinoma (SW480). To assess functionality, HAS3 expression in SW620 cells was inhibited by transfection with an asHAS3 construct. Decreased HA secretion and cell-surface retention by asHAS3 transfectants were confirmed using competitive binding and particle exclusion assays. Anchorage-independent growth, a correlate of tumor growth in vivo, was assessed by colony formation in soft agar. SW620 cells stably transfected with asHAS3 demonstrated significant growth inhibition, as evidenced by fewer colonies and smaller colony area than either SW620 cells or cells transfected with vector alone. Addition of exogenous HA restored growth in asHAS3 transfectants. Thus, we demonstrate that pericellular HA secretion and retention and HAS3 expression are increased in metastatic colon carcinoma cells relative to cells derived from a primary tumor. Inhibition of HAS3 expression in these cells decreased the pericellular HA matrix and inhibited anchorage-independent growth. These data suggest that HA and HAS3 function in the growth and progression of colon carcinoma.


Incidence and predictors of multimorbidity in the elderly: a population-based longitudinal study.
We aimed to calculate 3-year incidence of multimorbidity, defined as the development of two or more chronic diseases in a population of older people free from multimorbidity at baseline. Secondly, we aimed to identify predictors of incident multimorbidity amongst life-style related indicators, medical conditions and biomarkers. Data were gathered from 418 participants in the first follow up of the Kungsholmen Project (Stockholm, Sweden, 1991-1993, 78+ years old) who were not affected by multimorbidity (149 had none disease and 269 one disease), including a social interview, a neuropsychological battery and a medical examination. After 3 years, 33.6% of participants who were without disease and 66.4% of those with one disease at baseline, developed multimorbidity: the incidence rate was 12.6 per 100 person-years (95% CI: 9.2-16.7) and 32.9 per 100 person-years (95% CI: 28.1-38.3), respectively. After adjustments, worse cognitive function (OR, 95% CI, for 1 point lower Mini-Mental State Examination: 1.22, 1.00-1.48) was associated with increased risk of multimorbidity among subjects with no disease at baseline. Higher age was the only predictor of multimorbidity in persons with one disease at baseline. Multimorbidity has a high incidence at old age. Mental health-related symptoms are likely predictors of multimorbidity, suggesting a strong impact of mental disorders on the health of older people.


Long-term immunosuppression, without maintenance prednisone, after kidney transplantation.
Concern exists that prednisone-free maintenance immunosuppression in kidney transplant recipients will increase acute and/or chronic rejection. From October 1, 1999, through February 29, 2004, at our center, 477 kidney transplant recipients (341 living donor, 136 cadaver) discontinued prednisone on postoperative day 6, per our protocol. Immunosuppression consisted of polyclonal antibody (Thymoglobulin) for 5 days, prednisone intraoperatively and for 5 days, a calcineurin inhibitor, and either sirolimus or mycophenolate mofetil. We compared outcome with that of historical controls who did not discontinue prednisone. The recipients on prednisone-free maintenance immunosuppression had excellent 4-year actuarial patient survival (92%), graft survival (90%), acute rejection-free graft survival (86%), and chronic rejection-free graft survival (95%). The mean serum creatinine level (+/- SD) at 1 year was 1.6 +/- 0.6; at 4 years, 1.6 +/- 0.6. We noted that 8% of recipients had cytomegalovirus (CMV) disease; 4.5%, fractures; 2.8%, cataracts; 1%, posttransplant diabetes; 0.2%, avascular necrosis; 0.2%, posttransplant lymphoproliferative disease; and 0%, polyomavirus. In all, 85% of kidney recipients with functioning grafts remain prednisone-free as of April 1, 2004. As compared with historical controls, the recipients on prednisone-free maintenance immunosuppression had better patient (P = 0.02) and graft survival (P < 0.0001) and lower rates of acute (P = 0.0004) and chronic (P = 0.02) rejection. In addition, they had a significantly lower rate of CMV disease (P < 0.0001), cataracts (P < 0.0001), posttransplant diabetes (P < 0.0001), and avascular necrosis (P = 0.0003). Prednisone-related side effects can be minimized without maintenance immunosuppression; our prednisone-free recipients do not have increased acute or chronic rejection.


Treatment of multiple system atrophy using intravenous immunoglobulin.
Multiple system atrophy (MSA) is a progressive neurodegenerative disorder of unknown etiology, manifesting as combination of parkinsonism, cerebellar syndrome and dysautonomia. Disease-modifying therapies are unavailable. Activation of microglia and production of toxic cytokines suggest a role of neuroinflammation in MSA pathogenesis. This pilot clinical trial evaluated safety and tolerability of intravenous immunoglobulin (IVIG) in MSA. This was a single-arm interventional, single-center, open-label pilot study. Interventions included monthly infusions of the IVIG preparation Privigen®, dose 0.4 gram/kg, for 6 months. Primary outcome measures evaluated safety and secondary outcome measures evaluated preliminary efficacy of IVIG. Unified MSA Rating Scale (UMSARS) was measured monthly. Quantitative brain imaging using 3T MRI was performed before and after treatment. Nine subjects were enrolled, and seven (2 women and 5 men, age range 55-64 years) completed the protocol. There were no serious adverse events. Systolic blood pressure increased during IVIG infusions (p<0.05). Two participants dropped out from the study because of a non-threatening skin rash. The UMSARS-I (activities of daily living) and USMARS-II (motor functions) improved significantly post-treatment. UMSARS-I improved in all subjects (pre-treatment 23.9 ± 6.0 vs. post-treatment 19.0±5.9 (p=0.01). UMSARS-II improved in 5 subjects, was unchanged in 1 and worsened in 1 (pre-treatment 26.1±7.5 vs. post-treatment 23.3±7.3 (p=0.025). The MR imaging results were not different comparing pre- to post-treatment. Treatment with IVIG appears to be safe, feasible and well tolerated and may improve functionality in MSA. A larger, placebo-controlled study is needed.


Influence of thermal processing on hydrolysis and stability of folate poly-gamma-glutamates in broccoli (Brassica oleracea var. italica), carrot (Daucus carota) and tomato (Lycopersicon esculentum).
The folate poly-gamma-glutamate profile, their concentrations, and hydrolysis by endogenous gamma-glutamyl hydrolase (GGH) were evaluated in broccoli, carrot and tomato. Further studies on the effect of time and temperature on folate poly-gamma-glutamate hydrolysis and stability were carried out in broccoli since this vegetable showed the highest long-chain and total folate poly-gamma-glutamate concentration. The evolution of l-ascorbic acid, total phenols and Trolox equivalent antioxidant capacity (TEAC) values was evaluated in parallel. Upon thermal inactivation of GGH prior to crushing, it was observed that broccoli, carrot and tomato contained poly-gamma-glutamates with one to seven glutamate residues but differed in the predominant poly-gamma-glutamates. Crushing of raw broccoli, carrot and tomato resulted in significant poly-gamma-glutamate profile changes in broccoli and carrot (indicating GGH-catalyzed hydrolysis) but not in tomato. In this study, the actual crushing of raw broccoli matrix had a greater effect on folate poly-gamma-glutamate hydrolysis than incubation conditions (0-30 min at 25-55 degrees C). During treatments at 25-140 degrees C, folate retention was higher at 80 and 100 degrees C than at the other temperatures. A similar trend in thermal stability was observed for folates, vitamin C, total phenols and TEAC value, an indication that conditions that result in endogenous antioxidants degradation might also result in folate degradation.


Tandem RNA chimeras contribute to transcriptome diversity in human population and are associated with intronic genetic variants.
Chimeric RNAs originating from two or more different genes are known to exist not only in cancer, but also in normal tissues, where they can play a role in human evolution. However, the exact mechanism of their formation is unknown. Here, we use RNA sequencing data from 462 healthy individuals representing 5 human populations to systematically identify and in depth characterize 81 RNA tandem chimeric transcripts, 13 of which are novel. We observe that 6 out of these 81 chimeras have been regarded as cancer-specific. Moreover, we show that a prevalence of long introns at the fusion breakpoint is associated with the chimeric transcripts formation. We also find that tandem RNA chimeras have lower abundances as compared to their partner genes. Finally, by combining our results with genomic data from the same individuals we uncover intronic genetic variants associated with the chimeric RNA formation. Taken together our findings provide an important insight into the chimeric transcripts formation and open new avenues of research into the role of intronic genetic variants in post-transcriptional processing events.


Fra-1 governs cell migration via modulation of CD44 expression in human mesotheliomas.
Silencing of Fra-1, a component of the dimeric transcription factor, activator protein-1 (AP-1), inhibits mRNA expression of c-met and cd44 in rat mesothelioma cells and is causally linked to maintenance of the transformed phenotype. However, the mechanisms of Fra-1 regulation and Fra-1 regulated gene expression in human malignant mesothelioma (MM) are unclear. We first show in a panel of human MM cells that Fra-1 mRNA expression in MM is complex and regulated by extracellular signal-regulated kinase (ERK1, ERK2), Src, and phosphatidyl-inositol-3-kinase (PI3K) pathways in a tumor-specific fashion. Cell lines with PI3K-dependent Fra-1 expression were SV40 positive and expressed the lowest basal Fra-1 levels. Levels of Fra-1 expression correlated with amounts of CD44 expression that were greater in simian virus 40 negative (SV40-) MM cells. Using dominant negative (dn), short hairpin (sh) and small interference (si) RNA constructs, we next demonstrate that expression of CD44, the principal hyaluronic receptor in MMs, correlates with Fra-expression in both simian virus 40 positive (SV40+) and SV40- MMs. Moreover, both Fra-1 and CD44 expression are linked to cell migration in SV40- MM cells. Lastly, in contrast to normal lung tissue, tissue microarrays revealed that Fra-1 was expressed in 33 of 34 human MMs, and that all CD44+ tumors were SV40-. These results suggest that Fra-1 is associated with cell migration in human MMs and that Fra-1 modulation of CD44 may govern migration of selected MMs.


Cow-specific treatment of clinical mastitis: an economic approach.
Under Dutch circumstances, most clinical mastitis (CM) cases of cows on dairy farms are treated with a standard intramammary antimicrobial treatment. Several antimicrobial treatments are available for CM, differing in antimicrobial compound, route of application, duration, and cost. Because cow factors (e.g., parity, stage of lactation, and somatic cell count history) and the causal pathogen influence the probability of cure, cow-specific treatment of CM is often recommended. The objective of this study was to determine if cow-specific treatment of CM is economically beneficial. Using a stochastic Monte Carlo simulation model, 20,000 CM cases were simulated. These CM cases were caused by Streptococcus uberis and Streptococcus dysgalactiae (40%), Staphylococcus aureus (30%), or Escherichia coli (30%). For each simulated CM case, the consequences of using different antimicrobial treatment regimens (standard 3-d intramammary, extended 5-d intramammary, combination 3-d intramammary+systemic, combination 3-d intramammary+systemic+1-d nonsteroidal antiinflammatory drugs, and combination extended 5-d intramammary+systemic) were simulated simultaneously. Finally, total costs of the 5 antimicrobial treatment regimens were compared. Some inputs for the model were based on literature information and assumptions made by the authors were used if no information was available. Bacteriological cure for each individual cow depended on the antimicrobial treatment regimen, the causal pathogen, and the cow factors parity, stage of lactation, somatic cell count history, CM history, and whether the cow was systemically ill. Total costs for each case depended on treatment costs for the initial CM case (including costs for antibiotics, milk withdrawal, and labor), treatment costs for follow-up CM cases, costs for milk production losses, and costs for culling. Average total costs for CM using the 5 treatments were (US) $224, $247, $253, $260, and $275, respectively. Average probabilities of bacteriological cure for the 5 treatments were 0.53, 0.65, 0.65, 0.68, and 0.75, respectively. For all different simulated CM cases, the standard 3-d intramammary antimicrobial treatment had the lowest total costs. The benefits of lower costs for milk production losses and culling for cases treated with the intensive treatments did not outweigh the higher treatment costs. The stochastic model was developed using information from the literature and assumptions made by the authors. Using these information sources resulted in a difference in effectiveness of different antimicrobial treatments for CM. Based on our assumptions, cow-specific treatment of CM was not economically beneficial.


Proteome-wide prediction of bacterial carbohydrate-binding proteins as a tool for understanding commensal and pathogen colonisation of the vaginal microbiome.
Bacteria use carbohydrate-binding proteins (CBPs), such as lectins and carbohydrate-binding modules (CBMs), to anchor to specific sugars on host surfaces. CBPs in the gut microbiome are well studied, but their roles in the vagina microbiome and involvement in sexually transmitted infections, cervical cancer and preterm birth are largely unknown. We established a classification system for lectins and designed Hidden Markov Model (HMM) profiles for data mining of bacterial genomes, resulting in identification of >100,000 predicted bacterial lectins available at unilectin.eu/bacteria. Genome screening of 90 isolates from 21 vaginal bacterial species shows that those associated with infection and inflammation produce a larger CBPs repertoire, thus enabling them to potentially bind a wider array of glycans in the vagina. Both the number of predicted bacterial CBPs and their specificities correlated with pathogenicity. This study provides new insights into potential mechanisms of colonisation by commensals and potential pathogens of the reproductive tract that underpin health and disease states.


Informal m-health: How are young people using mobile phones to bridge healthcare gaps in Sub-Saharan Africa?
The African communications 'revolution' has generated optimism that mobile phones might help overcome infrastructural barriers to healthcare provision in resource-poor contexts. However, while formal m-health programmes remain limited in coverage and scope, young people are using mobile phones creatively and strategically in an attempt to secure effective healthcare. Drawing on qualitative and quantitative data collected in 2012-2014 from over 4500 young people (aged 8-25 y) in Ghana, Malawi and South Africa, this paper documents these practices and the new therapeutic opportunities they create, alongside the constraints, contingencies and risks. We argue that young people are endeavouring to lay claim to a digitally-mediated form of therapeutic citizenship, but that a lack of appropriate resources, social networks and skills ('digital capital'), combined with ongoing shortcomings in healthcare delivery, can compromise their ability to do this effectively. The paper concludes by offering tentative suggestions for remedying this situation.


Nearly degenerate four-wave mixing investigation of period doubling in semiconductor lasers.
The nearly degenerate four-wave mixing technique is used to investigate the physical origin of period doubling in the route toward chaos. This spectroscopic approach identifies resonantly enhanced nearly degenerate four-wave mixing processes as being responsible for the generation of subharmonic frequencies. The technique presented in the case of GaAlAs semiconductor lasers could be applied to the study of monomode instabilities occurring in any laser or bistable devices.


NaSrMn2F7, NaCaFe2F7, and NaSrFe2F7: novel single crystal pyrochlore antiferromagnets.
The crystal structures and magnetic properties of three previously unreported A2B2F7 pyrochlore materials, NaSrMn2F7, NaCaFe2F7, and NaSrFe2F7 are presented. In these compounds, either S = 2Fe2+ or S = 5/2Mn2+ is on the B site, while nonmagnetic Na and Ca (Na and Sr) are disordered on the A site. The materials, which were grown as crystals via the floating zone method, display high effective magnetic moments and large Curie-Weiss thetas. Despite these characteristics, no ordering transition is detected. However, freezing of the magnetic spins, characterized by peaks in the susceptibility or specific heat, is observed at very low temperatures. The empirical frustration index, f = -θ CW/T f, for the materials are 36 (NaSrMn2F7), 27 (NaSrFe2F7), and 19 (NaCaFe2F7). AC susceptibility, DC susceptibility, and heat capacity measurements are used to characterize the observed spin glass behavior. The results suggest that the compounds are frustrated pyrochlore antiferromagnets with weak bond disorder. The magnetic phenomena that these fluoride pyrochlores exhibit, in addition to their availability as relatively large single crystals, make them promising candidates for the study of geometric magnetic frustration.


Loss of interleukin 4 receptor-associated molecule gp200-MR6 in human breast cancer: prognostic significance.
Several in vitro studies stress a potentially important role of interleukin 4 (IL-4) and the related gp200-MR6 molecule in the immunological response to cancer and in tumour proliferation. In the present study, we assessed the expression of gp200-MR6 in primary breast cacrinomas using the MR6 monoclonal antibody. Results were correlated with tumour parameters (T-,N-stage, histology, grade, oestrogen and epidermal growth factor (EGF) receptors), and the impact on survival was assessed. Twenty-four out of 110 cases (22%) were positive for gp200-MR6, 62 out of 110 (56%) expressed weak staining and 24 out of 114 (22%) did not stain. The normal breast epithelia were invariably stained for gp200-MR6 showing that down-regulation or loss of this molecule occurred during the evolution of breast cancer. Gp200-MR6 loss was independent from differentiation, nodal positivity and oestrogen receptor levels as well as patients' age. Loss of the gp200-MR6 molecule was more frequent in lobular cases (P=0.03). The overall survival was better, although not reaching statistical significance, in patients with positive gp200-MR6 expression (92% alive at 5 years compared with 70% for those with weak or no expression, P=0.1). The local relapse-free survival was independent of gp200-MR6 status. It is concluded that loss of gp200-MR6 may be one of the mechanisms through which breast cancer cells escape immune surveillance, resulting in an increased metastatic potential and poorer outcome. Evidence of down-regulation of the gp200-MR6 molecule has implications for IL-4-linked toxin therapy and, as IL-4 is an inhibitor of breast epithelial growth, may represent loss of a tumour-suppression mechanism.


Analysis and Tools for Improved Management of Connectionless and Connection-Oriented BLE Devices Coexistence.
With the introduction of low-power wireless technologies, like Bluetooth Low Energy (BLE), new applications are approaching the home automation, healthcare, fitness, automotive and consumer electronics markets. BLE devices are designed to maximize the battery life, i.e., to run for long time on a single coin-cell battery. In typical application scenarios of home automation and Ambient Assisted Living (AAL), the sensors that monitor relatively unpredictable and rare events should coexist with other sensors that continuously communicate health or environmental parameter measurements. The former usually work in connectionless mode, acting as advertisers, while the latter need a persistent connection, acting as slave nodes. The coexistence of connectionless and connection-oriented networks, that share the same central node, can be required to reduce the number of handling devices, thus keeping the network complexity low and limiting the packet's traffic congestion. In this paper, the medium access management, operated by the central node, has been modeled, focusing on the scheduling procedure in both connectionless and connection-oriented communication. The models have been merged to provide a tool supporting the configuration design of BLE devices, during the network design phase that precedes the real implementation. The results highlight the suitability of the proposed tool: the ability to set the device parameters to allow us to keep a practical discovery latency for event-driven sensors and avoid undesired overlaps between scheduled scanning and connection phases due to bad management performed by the central node.


Inflammation-induced preterm birth alters neuronal morphology in the mouse fetal brain.
Adverse neurological outcome is a major cause of long-term morbidity in ex-preterm children. To investigate the effect of parturition and inflammation on the fetal brain, we utilized two in vivo mouse models of preterm birth. To mimic the most common human scenario of preterm birth, we used a mouse model of intrauterine inflammation by intrauterine infusion of lipopolysaccharide (LPS). To investigate the effect of parturition on the immature fetal brain, in the absence of inflammation, we used a non-infectious model of preterm birth by administering RU486. Pro-inflammatory cytokines (IL-10, IL-1beta, IL-6 and TNF-alpha) in amniotic fluid and inflammatory biomarkers in maternal serum and amniotic fluid were compared between the two models using ELISA. Pro-inflammatory cytokine expression was evaluated in the whole fetal brains from the two models. Primary neuronal cultures from the fetal cortex were established from the different models and controls in order to compare the neuronal morphology. Only the intrauterine inflammation model resulted in an elevation of inflammatory biomarkers in the maternal serum and amniotic fluid. Exposure to inflammation-induced preterm birth, but not non-infectious preterm birth, also resulted in an increase in cytokine mRNA in whole fetal brain and in disrupted fetal neuronal morphology. In particular, Microtubule-associated protein 2 (MAP2) staining was decreased and the number of dendrites was reduced (P < 0.001, ANOVA between groups). These results suggest that inflammation-induced preterm birth and not the process of preterm birth may result in neuroinflammation and alter fetal neuronal morphology.


Quebec platelet disorder.
A case of Quebec platelet disorder is hereby reported. A 33 years old woman presented with history of epistaxis and gum bleeding since childhood and menorrhagia and bleeding per vaginum after puberty, also had history of excessive blood loss after birth of child. Her coagulation profile was normal but platelet function testing by platelet aggregation assay showed abnormal aggregation of platelet with epinephrine. This type of response is seen in "Quebec platelet disorder" which is a rare autosomal dominant disorder of platelet function characterized by increased bleeding after any injury or trauma.


Ultra-high performance liquid chromatography-size exclusion chromatography (UPLC-SEC) as an efficient tool for the rapid and highly informative characterisation of biopolymers.
Starch has a complex molecular structure, with properties dependent on the relative chain lengths and branching structure of its constituent molecules, which varies due to polymorphisms in starch biosynthetic genes, as well as environmental factors. Here we present the application of ultra-high performance size exclusion chromatography to the separation of starch chains from plant seeds. Several methods, have been used to analyse chain length distributions in starch, all with limitations in terms of analysis time, sample preparation and molecular weight range. Here we demonstrate that chain length distributions can be obtained with dramatically reduced analysis time using ultra-high performance size exclusion chromatography. The method may also show improvements in resolution of some fine structural features. Understanding links between starch fine structure and biosynthetic genes will allow bioengineering of starches with tailored properties. This technique may have application to the size separation and resolution of a range of biopolymers of value to the food, drink and pharmaceutical industries.


HIV-1 protease inhibitor induced oxidative stress suppresses glucose stimulated insulin release: protection with thymoquinone.
The highly active anti-retroviral therapy (HAART) regimen has considerably reduced the mortality rate in HIV-1 positive patients. However, long-term exposure to HAART is associated with a metabolic syndrome manifesting cardiovascular dysfunction, lipodystrophy, and insulin resistance syndrome (IRS). The inclusion of HIV-1 protease inhibitors (PIs) in HAART has been linked to the induction of IRS. Although several molecular mechanisms of PI-induced effects on insulin action have been postulated, the deleterious effects of PIs on insulin production by pancreatic beta-cells have not been fully investigated and therapeutic strategies to ameliorate insulin dysregulation at this level have not been targeted. The present study showed that exposure to several different PIs, nelfinavir (5-10 microM), saquinavir (5-10 microM) and atazanavir (8-20 microM), decreases glucose stimulated insulin secretion from rat pancreatic beta-cells (INS-1). Nelfinavir significantly increased reactive oxygen species (ROS) generation and suppressed cytosolic, but not mitochondrial superoxide dismutase (SOD) levels. Nelfinvair also decreased both glutathione and ATP and increased UCP2 levels in these cells. Simultaneous treatment with thymoquinone (TQ) (2.5 microM), an active ingredient of black seed oil, significantly inhibited the effect of nelfinavir on augmented ROS production and suppressed SOD levels. Both TQ and black seed oil exposure increased glucose stimulated insulin secretion and ameliorated the suppressive effect of nelfinavir. The present findings imply a direct role of ROS in PI induced deleterious effects on pancreatic beta-cells. Our findings also suggest that TQ may be used as a potential therapeutic agent to normalize the dysregulated insulin production observed in HAART treated patients.


Comparative susceptibility of mosquito species and strains to oral and parenteral infection with dengue and Japanese encephalitis viruses.
Thirty-four strains of Asian and Pacific mosquitoes belonging to 22 species of 7 genera were compared for oral and/or parenteral susceptibility to infection with 1 or more strains of each of the 4 dengue serotypes. Surprisingly, several species of common man-biting Aedes were much more susceptible to oral infection with each of the 4 dengue serotypes than was Aedes aegypti. These species included Aedes albopictus and members of the scutellaris group of the subgenus Stegomyia found on South Pacific islands. Mosquito strains and species relatively susceptible to 1 dengue serotype usually were relatively susceptible to the others also. Almost all species of Aedes tested were uniformly susceptible to parenteral infection with the dengue viruses but, with the exception of a species of Tripteroides, species of all other genera were comparatively resistant to that mode of infection. Dengue viruses usually replicated to about the same extent in orally-infected mosquitoes as they did in parenterally-infected specimens of the same species. Seventeen species of mosquitoes of 7 genera also were tested for parenteral susceptibility to infection with Japanese encephalitis virus. With the possible exception of 2 species of Anopheles, the virus replicated to about the same degree in all species tested and achieved levels considerably higher than did any of the dengue viruses in the same mosquito strain and species held under the same conditions.


Nucleotide sequence of the long terminal repeat and flanking cellular sequences of avian endogenous retrovirus ev-2: variation in Rous-associated virus-0 expression cannot be explained by differences in primary sequence.
A fragment of chicken DNA containing the left long terminal repeat of endogenous retrovirus ev-2 and flanking cellular sequences has been molecularly cloned and analyzed. Comparison with sequence data from the analogous regions of ev-1 and Rous-associated virus-0 viral DNA reveals similarities among flanking regions of the integrated proviruses and among all three long terminal repeats. From the latter finding, we conclude that the difference in level of expression of ev-2 and its progeny Rous-associated virus-0 provirus cannot be due to sequence differences in their upstream long terminal repeats.


Better than men? Sex and the therapy/enhancement distinction.
The normative significance of the distinction between therapy and enhancement has come under sustained philosophical attack in recent discussions of the ethics of shaping future persons by means of advanced genetic technologies. Giving up the idea that whether a condition is normal or not should play a crucial role in assessing the ethics of genetic interventions has unrecognized and strongly counterintuitive implications when it comes to selecting what sort of children should be brought into the world. According to standard philosophical accounts of the factors one should take into account when making such decisions, women are "better than men." Given the biological differences between the sexes, then, if the only concern is the capacities of an embryo rather than its capacities relative to some normatively significant baseline, there is compelling reason to choose only female embryos. In order to avoid this radical and counterintuitive conclusion, one must embrace the idea that both sexes are normal. The strength of the prima facie reasons to select or reject embryos depends on their sex, which is to say that it depends on the normal capacities of their sex. The therapy/enhancement distinction therefore plays a crucial role in determining the ethics of interventions into the genetics of future generations.


Preschool attachment, self-esteem and the development of preadolescent anxiety and depressive symptoms.
This study examined the longitudinal association between preschool attachment patterns, the development of anxiety and depression at preadolescence and the mediational role of self-esteem. Child-mother attachment classifications of 68 children (33 girls) were assessed between 3-4 years of age (M = 3.7 years, SD = 4.4 months) using the Separation-Reunion Procedure. At age 11-12 (M = 11.7 years, SD = 4.3 months), anxiety and depressive symptoms (Dominic Interactive Questionnaire), and self-esteem (Self-Perception Profile for Children) were also evaluated. Preadolescents who had shown disorganized attachment at preschool age scored higher on both anxiety and depression and lower on self-esteem than those who had shown secure and insecure-organized attachment strategies. Self-esteem was a partial mediator of the association between preschool disorganization and symptoms of preadolescent depression, but the model was not supported for anxiety. These findings support the idea that early attachment and self-esteem should be central themes in prevention programs with young children.


Parity-time symmetric coupled microresonators with a dispersive gain/loss.
The paper reports on the coupling of Parity-Time (PT)-symmetric whispering gallery resonators with realistic material and gain/loss models. Response of the PT system is analyzed for the case of low and high material and gain dispersion, and also for two practical scenarios when the pump frequency is not aligned with the resonant frequency of the desired whispering gallery mode and when there is imbalance in the gain/loss profile. The results show that the presence of dispersion and frequency misalignment causes skewness in frequency bifurcation and significant reduction of the PT breaking point, respectively. Finally, we demonstrate a lasing mode operation which occurs due to an early PT-breaking by increasing loss in a PT system with unbalanced gain and loss.


CUDASW++ 3.0: accelerating Smith-Waterman protein database search by coupling CPU and GPU SIMD instructions.
The maximal sensitivity for local alignments makes the Smith-Waterman algorithm a popular choice for protein sequence database search based on pairwise alignment. However, the algorithm is compute-intensive due to a quadratic time complexity. Corresponding runtimes are further compounded by the rapid growth of sequence databases. We present CUDASW++ 3.0, a fast Smith-Waterman protein database search algorithm, which couples CPU and GPU SIMD instructions and carries out concurrent CPU and GPU computations. For the CPU computation, this algorithm employs SSE-based vector execution units as accelerators. For the GPU computation, we have investigated for the first time a GPU SIMD parallelization, which employs CUDA PTX SIMD video instructions to gain more data parallelism beyond the SIMT execution model. Moreover, sequence alignment workloads are automatically distributed over CPUs and GPUs based on their respective compute capabilities. Evaluation on the Swiss-Prot database shows that CUDASW++ 3.0 gains a performance improvement over CUDASW++ 2.0 up to 2.9 and 3.2, with a maximum performance of 119.0 and 185.6 GCUPS, on a single-GPU GeForce GTX 680 and a dual-GPU GeForce GTX 690 graphics card, respectively. In addition, our algorithm has demonstrated significant speedups over other top-performing tools: SWIPE and BLAST+. CUDASW++ 3.0 is written in CUDA C++ and PTX assembly languages, targeting GPUs based on the Kepler architecture. This algorithm obtains significant speedups over its predecessor: CUDASW++ 2.0, by benefiting from the use of CPU and GPU SIMD instructions as well as the concurrent execution on CPUs and GPUs. The source code and the simulated data are available at http://cudasw.sourceforge.net.


Hearing gestures, seeing music: vision influences perceived tone duration.
Percussionists inadvertently use visual information to strategically manipulate audience perception of note duration. Videos of long (L) and short (S) notes performed by a world-renowned percussionist were separated into visual (Lv, Sv) and auditory (La, Sa) components. Visual components contained only the gesture used to perform the note, auditory components the acoustic note itself. Audio and visual components were then crossed to create realistic musical stimuli. Participants were informed of the mismatch, and asked to rate note duration of these audio-visual pairs based on sound alone. Ratings varied based on visual (Lv versus Sv), but not auditory (La versus Sa) components. Therefore while longer gestures do not make longer notes, longer gestures make longer sounding notes through the integration of sensory information. This finding contradicts previous research showing that audition dominates temporal tasks such as duration judgment.


An online neural substrate for sense of agency.
"Sense of agency" refers to the feeling of controlling an external event through one's own action. On one influential view, sense of agency is inferred after an action, by "retrospectively" comparing actual effects of actions against their intended effects. In contrast, a "prospective" component of agency, generated during action selection, and in advance of knowing the actual effect, has received less attention. Here, we used functional magnetic resonance imaging to investigate prospective contributions of action selection processes to sense of agency. To do so, we dissociated action selection processes from action-outcome matching, by subliminally priming responses to a target. We found that participants experienced greater control over action effects when the action was compatibly versus incompatibly primed. Thus, compatible primes facilitated action selection processing, in turn boosting sense of agency over a subsequent effect. This prospective contribution of action selection processes to sense of agency was accounted for by exchange of signals across a prefrontal-parietal network. Specifically, we found that the angular gyrus (AG) monitors signals relating to action selection in dorsolateral prefrontal cortex, to prospectively inform subjective judgments of control over action outcomes. Online monitoring of these signals by AG might provide the subject with a subjective marker of volition, prior to action itself.


Analysis of copy number variation in Alzheimer's disease in a cohort of clinically characterized and neuropathologically verified individuals.
Copy number variations (CNVs) are genomic regions that have added (duplications) or deleted (deletions) genetic material. They may overlap genes affecting their function and have been shown to be associated with disease. We previously investigated the role of CNVs in late-onset Alzheimer's disease (AD) and mild cognitive impairment using Alzheimer's Disease Neuroimaging Initiative (ADNI) and National Institute of Aging-Late Onset AD/National Cell Repository for AD (NIA-LOAD/NCRAD) Family Study participants, and identified a number of genes overlapped by CNV calls. To confirm the findings and identify other potential candidate regions, we analyzed array data from a unique cohort of 1617 Caucasian participants (1022 AD cases and 595 controls) who were clinically characterized and whose diagnosis was neuropathologically verified. All DNA samples were extracted from brain tissue. CNV calls were generated and subjected to quality control (QC). 728 cases and 438 controls who passed all QC measures were included in case/control association analyses including candidate gene and genome-wide approaches. Rates of deletions and duplications did not significantly differ between cases and controls. Case-control association identified a number of previously reported regions (CHRFAM7A, RELN and DOPEY2) as well as a new gene (HLA-DRA). Meta-analysis of CHRFAM7A indicated a significant association of the gene with AD and/or MCI risk (P = 0.006, odds ratio = 3.986 (95% confidence interval 1.490-10.667)). A novel APP gene duplication was observed in one case sample. Further investigation of the identified genes in independent and larger samples is warranted.


[Clinical observation on post-stroke shoulder pain treated with balance acupuncture].
To observe the difference of therapeutic effect on post-stroke shoulder pain between balance acupuncture and conventional acupuncture. Sixty cases were randomly divided into two groups, 30 cases in each one. In balance acupuncture group, the balance technique of acupuncture was used on Jiantong point. In conventional acupuncture group, the conventional acupuncture was applied to Shuigou (GV 26), Neiguan (PC 6), Tongli (HT 5), Zusanlti (ST 36), Xuanzhong (GB 39) and others, combined with the points on the affected meridians. The Visual Analogue Scale (VAS) was adopted to evaluate the efficacy. Separately, before and after the 1st treatment, VAS scores were 5.62 +/- 0.46 and 5.00 +/- 0.31 in balance acupuncture group (P < 0.01), and those were 5.52 +/- 0.65 and 5.22 +/- 0.29 in conventional acupuncture group (P < 0.05). The improvement in balance acupuncture group was superior to that in conventional acupuncture group (P < 0.05). After 10 treatments (one course of treatment), VAS score was 3.75 +/- 0.67 in balance acupuncture group and was 3.77 +/- 0.62 in conventional acupuncture group, which reduced significantly as compared with those before treatment (both P < 0.01), the difference between two groups was not significant (P > 0.05). Either balance acupuncture or conventional acupuncture has significant analgesia on post-stroke shoulder pain. But, the instant analgesia of balance acupuncture is superior to that of conventional acupuncture.


Demodex Species Infestation in Patients with Ear Itching and Its Relationship to Itch Severity.
The aim of the present study was to investigate Demodex species infestation in patients with ear itching. The relationship between the severity of ear itching and Demodex spp. positivity has not been previously reported in the literature, and we believe that our study will make a significant contribution to the understanding of the etiology of ear itching. Fifty patients with itching ears and 54 healthy control patients were asked to use a visual analogue scale (VAS) to rate the itch, the itching period, and the medication used for the itching. All samples were evaluated for Demodex spp. under a light microscope. There was no statistically significant difference between the groups in terms of numbers of Demodex spp. (p=0.154), and there was no statistically significant difference between the groups in terms of Demodex spp. positivity (p=0.054). Despite the lack of statistically significant differences, Demodex spp. infestations were more common in the affected group than in the control group. A positive and strongly significant relationship was observed between the number of Demodex spp. and severity of ear itch in the patient group based on VAS scores (p=0.0001; r=0.724). We found that an increased number of Demodex spp. was strongly related to increased severity of ear itching.


Pearls and Pitfalls in Diagnosing Pediatric Urinary Bladder Masses.
Urinary bladder masses are rare in children, and the associated histologic features and prognoses in this population are different from those in adults. Most children with urinary bladder masses present with lower urinary tract symptoms, which may include hematuria, dysuria, frequent urination, and urgency to urinate. However, some of these masses may be identified incidentally or involve generic symptoms such as abdominal distention. In general, pediatric bladder tumors can be divided into those that originate from the bladder epithelium, known as urothelial neoplasms, and mesenchymal bladder neoplasms, which are more prevalent. The most common bladder malignancy in children is a rhabdomyosarcoma, whereas the most common benign bladder lesion in the pediatric population is a papillary urothelial neoplasm of low malignant potential (PUNLMP). The first-line imaging tool for assessing bladder lesions is ultrasonography, which may be followed by a cross-sectional imaging examination such as computed tomography or magnetic resonance imaging if the origin of the mass is unclear or if distant spread is suspected. Although imaging may enable the radiologist to suggest a differential diagnosis based on lesion location and patient age, tissue biopsy generally is required to identify the exact pathologic entity. This is usually performed at cystoscopy and may be curative in cases in which the lesion is small and has low recurrence potential. Knowledge of the clinical, histopathologic, and imaging features of common bladder neoplasms is essential, as it can aid in preventing imaging pitfalls. These may include the misinterpretation of either a pelvic mass as arising from the bladder or a bladder mass as arising from the pelvis, and interpreting an inflammatory mass or bladder detritus as a neoplasm. ©RSNA, 2017.


First Report of Tagetes erecta Damping Off Caused by Ceratobasidium sp. from India.
The Mexican marigold (Tagetes erecta) is cultivated commercially in India for medicinal, ceremonial, and decorative purposes. It is native to Mexico and the United States. The natural phytochemical thiophene extracted from T. erecta has been shown to have antibacterial activity. It is also grown to extract lutein, a common yellow/orange food color. During winter of 2011, approximately 15% marigold seedlings exhibited damping off symptoms at CSIR-CIMAP, Lucknow, India, and its adjoining areas. Infected seedlings initially produced water-soaked lesions on the stem at the soil level that later turned pinkish with a brownish halo in the center. The infected seedlings were cut into small pieces, surface disinfected with 1% sodium hypochlorite, rinsed thrice with sterile distilled water, and placed onto potato dextrose agar (PDA) plates. The plates were incubated at 25°C for 3 days. The isolation yielded whitish fungal growth that later turned tan brown. The mycelium was binucleate, septate, sub branching at right angles, with distinct constriction at the origin of branching. Bisbenzamide (Hoechst 33258; Chemical Abstracts no. 23491-45-4) was used as fluorescent dye for the staining of nuclei. Based on cultural as well as morphological characteristic features, the fungus was identified as Ceratobasidium sp. (1,2). The molecular identification was on the basis of internal transcribed spacer (ITS) region sequence. Amplification of the ITS of rDNA using primers ITS1/ITS4 yielded a ~700 bp band and sequenced data were deposited in the NCBI GenBank (KC193238). The ITS region (700 bp) was 100% identical to Ceratobasidium sp. AG-F strain BAGF101 isolated from Musa spp. in Georgia, United States (GenBank Accession No. HQ168370). The pathogenicity of the fungus was tested under glasshouse conditions. The inoculum of the fungus was prepared on sterile maize seeds in Erlenmeyer flasks by inoculating seeds with three disks (1 mm) of 5-day-old culture, and kept at 25 ± 2 °C for 14 days in the dark. The healthy, 5 to 7 day old seedlings were inoculated with five artificially infested maize seeds per pot. The uninoculated seedlings served as control. Both inoculated and uninoculated seedlings were kept at 28 ± 2°C in a humidity (95%) chamber for 3 days and thereafter placed in the glasshouse at 28 ± 2°C for development of disease symptoms. Initial symptoms developed as water-soaked lesions on the infected seedlings in 2 to 3 days, while typical disease symptoms appeared after 4 to 5 days of inoculation. Uninoculated seedlings were free from infection. The fungus was reisolated from the artificially infected seedlings on PDA and its identification as Ceratobasidium sp. was confirmed by morphological and molecular characteristics. Recently, Ceratobasidium sp. was reported as causal organism of root rot on Atractylodes macrocephala and banana (3,4). To the best of our knowledge, marigold damping off disease caused by Ceratobasidium sp. has not been reported so far on T. erecta. Hence, it is the first report from India. During fungal disease management for marigold, association of Ceratobasidium sp. should not be ignored for better crop protection. References: (1) R. T. Moore. Mycotaxon 29:91, 1987. (2) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St. Paul, MN. 1991. (3) J. Yin et al. Plant Dis. 95:490, 2011. (4) J. M. You et al. Plant Dis.97:139, 2013.


Hypoxia inducible factor prolyl hydroxylases in inflammatory bowel disease.
Inflammatory bowel disease (IBD) is a chronic disease that is characterized by intestinal inflammation. Epithelial damage and loss of intestinal barrier function are believed to be the hallmark pathologies of the disease. In IBD, the resident and infiltrating immune cells consume much oxygen, rendering the inflamed intestinal mucosa hypoxic. In hypoxia, the hypoxia-inducible factor (HIF) is induced to cope with the lack of oxygen and protect intestinal barrier. Protein stability of HIF is tightly controlled by prolyl hydroxylases (PHDs). Stabilization of HIF through inhibition of PHDs is appearing as a new strategy of IBD treatment. Studies have shown that PHD-targeting is beneficial to the treatment of IBD. In this Review, we summarize the current understanding of the role of HIF and PHDs in IBD and discuss the therapeutic potential of targeting PHD-HIF pathway for IBD treatment.


Cold Pool-Land Surface Interactions in a Dry Continental Environment.
Cold pools influence convective initiation and organization, dust lofting, and boundary layer properties, but little is known about their interactions with the land surface, particularly in dry continental environments. In this study, two-way cold pool-land surface interactions are investigated using high-resolution idealized simulations of an isolated, transient cold pool evolving in a dry convective boundary layer. Results using a fully interactive land surface demonstrate that sensible heat fluxes are suppressed at the center of the cold pool but enhanced at the edge due to the spatial patterns of land surface cooling and the air temperature and wind speed perturbations. This leads to cold pool dissipation from the edge inward. Latent heat fluxes are primarily suppressed within the cold pool, and the magnitude of this suppression is controlled by competition between atmospheric and land surface effects. By comparing the fully interactive land surface simulation to a simulation with imposed surface fluxes, the land surface-cold pool feedbacks are shown to reduce the cold pool lifetime, extent, and intensity by up to 50% and influence the pattern of boundary layer turbulent kinetic energy recovery, which have significant implications for cold pool-induced convective initiation.


Modeling a student's behavior in a tutorial-like system using learning automata.
This paper presents a new philosophy to model the behavior of a student in a tutorial- like system using learning automata (LAs). The model of the student in our system is inferred using a higher level LA, referred to as a meta-LA , which attempts to characterize the learning model of the students (or student simulators), while the latter use the tutorial-like system. The meta-LA , in turn, uses LAs as a learning mechanism to try to determine if the student in question is a fast, normal, or slow learner. The ultimate long-term goal of the exercise is the following: if the tutorial- like system can understand how the student perceives and processes knowledge, it will be able to customize the way by which it communicates the knowledge to the student to attain an optimal teaching strategy. The proposed meta-LA scheme has been tested for numerous environments, including the established benchmarks, and the results obtained are remarkable. Indeed, to the best of our knowledge, this is the first published result that infers the learning model of an LA when it is externally treated as a black box, whose outputs are the only observable quantities. Additionally, our paper represents a new class of multiautomata systems, where the meta-LA synchronously communicates with the students, also modeled using LAs. The meta-LA's environment "observes" the progress of the student LA, and the response of the latter to the meta-LA actions is based on these observations. This paper also discusses the learning system implications of such a meta-LA.


Sociotechnical changing in healthcare.
This paper discusses a conceptual approach to the study of the implementation of ICTs in healthcare organizations. The paper uses some fundamental concepts from sociotechnical studies to address the complex process of change--the changing--that accompanies ICT innovations. The paper argues for the importance of the perspective of changing as a way to account for the dynamics as technology and people, organizations and institutions co-constitutively work-out their future together.


The Role of Electronic Learning in Orthopaedic Graduate Medical Training: A Consensus From Leaders in Orthopaedic Training Programs.
The US orthopaedic graduate medical education system is based on long established methods in education, but academic leaders at orthopaedic institutions now have the ability to use electronic learning innovations. Hospital for Special Surgery gathered graduate medical education leaders from orthopaedic training programs around the country and an electronic learning expert to review current orthopaedic residency and fellowship program practices. This group came to consensus with the following points: (1) current training methods do not take full advantage of available technology/innovations, (2) trainees inappropriately use electronic resources in the absence of or in an underdeveloped formal electronic training program, (3) trainees learn at different rates and in different ways requiring individualized plans for optimal content engagement, and (4) formal electronic learning programs better use time dedicated to educating trainees than informal programs. Orthopaedic graduate medical training programs that adopt these points can establish an electronic learning program to complement their traditional education program by (1) guaranteeing online content is standardized and approved, (2) reducing time spent covering standard lecture material and increasing time spent reviewing cases, and (3) engaging students of all learning backgrounds with content in both asynchronous and synchronous formats.


Cerebellar synaptic defects and abnormal motor behavior in mice lacking alpha- and beta-dystrobrevin.
The dystrobrevins (alphaDB and betaDB) bind directly to dystrophin and are components of a transmembrane dystrophin-glycoprotein complex (DGC) that links the cytoskeleton to extracellular proteins in many tissues. We show here that alphaDB, betaDB, and dystrophin are all concentrated at a discrete subset of inhibitory synapses on the somata and dendrites of cerebellar Purkinje cells. Dystrophin is depleted from these synapses in mice lacking both alphaDB and betaDB, and DBs are depleted from these synapses in mice lacking dystrophin. In dystrophin mutants and alphaDB,betaDB double mutants, the size and number of GABA receptor clusters are decreased at cerebellar inhibitory synapses, and sensorimotor behaviors that reflect cerebellar function are perturbed. Synaptic and behavioral abnormalities are minimal in mice lacking either alphaDB or betaDB. Together, our results show that the DGC is required for proper maturation and function of a subset of inhibitory synapses, that DB is a key component of this DGC, and that interference with this DGC leads to behavioral abnormalities. We suggest that motor deficits in muscular dystrophy patients, which are their cardinal symptoms, may reflect not only peripheral derangements but also CNS defects.


Compact ultrafast semiconductor disk laser: targeting GFP based nonlinear applications in living organisms.
We present a portable ultrafast Semiconductor Disk Laser (SDL) (or vertical extended cavity surface emitting laser-VECSELs), to be used for nonlinear microscopy. The SDL is modelocked using a quantum-dot semiconductor saturable absorber mirror (SESAM), delivering an average output power of 287 mW, with 1.5 ps pulses at 500 MHz and a central wavelength of 965 nm. Specifically, despite the fact of having long pulses and high repetition rates, we demonstrate the potential of this laser for Two-Photon Excited Fluorescence (TPEF) imaging of in vivo Caenorhabditis elegans (C. elegans) expressing Green Fluorescent Protein (GFP) in a set of neuronal processes and cell bodies. Efficient TPEF imaging is achieved due to the fact that this wavelength matches the peak of the two-photon action cross section of this widely used fluorescent marker. The SDL extended versatility is shown by presenting Second Harmonic Generation images of pharynx, uterus, body wall muscles and its potential to be used to excite other different commercial dyes. Importantly this non-expensive, turn-key, compact laser system could be used as a platform to develop portable nonlinear bio-imaging devices.


On the formation of anions: frequency-, angle-, and time-resolved photoelectron imaging of the menadione radical anion.
Frequency-, angle-, and time-resolved photoelectron imaging of gas-phase menadione (vitamin K3) radical anions was used to show that quasi-bound resonances of the anion can act as efficient doorway states to produce metastable ground electronic state anions on a sub-picosecond timescale. Several anion resonances have been experimentally observed and identified with the assistance of ab initio calculations, and ground state anion recovery was observed across the first 3 eV above threshold. Time-resolved measurements revealed the mechanism of electronic ground state anion formation, which first involves a cascade of very fast internal conversion processes to a bound electronic state that, in turn, decays by slower internal conversion to the ground state. Autodetachment processes from populated resonances are inefficient compared with electronic relaxation through internal conversion. The mechanistic understanding gained provides insight into the formation of radical anions in biological and astrochemical systems.


Partial lipodystrophy and insulin resistant diabetes in a patient with a homozygous nonsense mutation in CIDEC.
Lipodystrophic syndromes are characterized by adipose tissue deficiency. Although rare, they are of considerable interest as they, like obesity, typically lead to ectopic lipid accumulation, dyslipidaemia and insulin resistant diabetes. In this paper we describe a female patient with partial lipodystrophy (affecting limb, femorogluteal and subcutaneous abdominal fat), white adipocytes with multiloculated lipid droplets and insulin-resistant diabetes, who was found to be homozygous for a premature truncation mutation in the lipid droplet protein cell death-inducing Dffa-like effector C (CIDEC) (E186X). The truncation disrupts the highly conserved CIDE-C domain and the mutant protein is mistargeted and fails to increase the lipid droplet size in transfected cells. In mice, Cidec deficiency also reduces fat mass and induces the formation of white adipocytes with multilocular lipid droplets, but in contrast to our patient, Cidec null mice are protected against diet-induced obesity and insulin resistance. In addition to describing a novel autosomal recessive form of familial partial lipodystrophy, these observations also suggest that CIDEC is required for unilocular lipid droplet formation and optimal energy storage in human fat.


Post-partum hemorrhage in women with rare bleeding disorders.
Post-partum hemorrhage (PPH) accounts for a substantial fraction of maternal deaths in the general population. Among all women, however, those affected with rare bleeding disorders (RBDs) represent a particular group since to usual bleeding symptoms, they are likely to experience bleedings associated to obstetrical and gynaecological problems. Pregnancy and childbirth, two important stages in the life of a woman, pose a special clinical challenge in women with RBDs, since information about these issues are really scarce and limited to few case reports. These data show that all women with RBDs, except for FXI deficiency, have to be considered potentially at risk for developing PPH, therefore they should be monitored carefully during and immediately after pregnancy. The implication is that women with bleeding disorders may require prophylaxis and/or close observation for several weeks and should be followed by a multidisciplinary team including expertises such as laboratory haematologist, obstetrician-gynaecologist, anaesthesiologist, family physician, and laboratory technician.


Reducing Sweeping Frequencies in Microwave NDT Employing Machine Learning Feature Selection.
Nondestructive Testing (NDT) assessment of materials' health condition is useful for classifying healthy from unhealthy structures or detecting flaws in metallic or dielectric structures. Performing structural health testing for coated/uncoated metallic or dielectric materials with the same testing equipment requires a testing method that can work on metallics and dielectrics such as microwave testing. Reducing complexity and expenses associated with current diagnostic practices of microwave NDT of structural health requires an effective and intelligent approach based on feature selection and classification techniques of machine learning. Current microwave NDT methods in general based on measuring variation in the S-matrix over the entire operating frequency ranges of the sensors. For instance, assessing the health of metallic structures using a microwave sensor depends on the reflection or/and transmission coefficient measurements as a function of the sweeping frequencies of the operating band. The aim of this work is reducing sweeping frequencies using machine learning feature selection techniques. By treating sweeping frequencies as features, the number of top important features can be identified, then only the most influential features (frequencies) are considered when building the microwave NDT equipment. The proposed method of reducing sweeping frequencies was validated experimentally using a waveguide sensor and a metallic plate with different cracks. Among the investigated feature selection techniques are information gain, gain ratio, relief, chi-squared. The effectiveness of the selected features were validated through performance evaluations of various classification models; namely, Nearest Neighbor, Neural Networks, Random Forest, and Support Vector Machine. Results showed good crack classification accuracy rates after employing feature selection algorithms.


Multiple mechanisms of endocannabinoid response initiation in hippocampus.
Endocannabinoids (eCBs) act as retrograde messengers at inhibitory synapses of the hippocampal CA1 region. Current models place eCB synthesis in the postsynaptic pyramidal cell and the site of eCB action at cannabinoid receptors located on presynaptic interneuron terminals. Four responses at the CA1-interneuron synapse are attributed to eCBs: depolarization-induced suppression of inhibition (DSI), G-protein-coupled receptor-mediated enhancement of DSI (DeltaDSI), persistent suppression of evoked inhibitory postsynaptic currents (eIPSCs), and finally, mGluR-dependent long-term depression (iLTD). It has been proposed that all are mediated by the eCB, 2-arachidonoyl glycerol, yet there is evidence that DSI does not arise from the same underlying biochemical processes as the other responses. In view of the increasing importance of eCB effects in the brain, it will be essential to understand the mechanisms by which eCB effects are produced. Our results reveal new differences in the biochemical pathways by which the eCB-dependent responses are initiated. Both U73122, a phospholipase C antagonist, and RHC-80267, a diacylglycerol (DAG) lipase antagonist, prevented eCB-dependent iLTD induction by 3,5-dihydroxyphenylglycine (DHPG). However, mAChR activation does not cause eCB-dependent iLTD. Neither enzyme inhibitor affects DSI, and persistent eCB-dependent eIPSC suppression induced by either mGluRs or mAChRs is unaffected by U73122. On the other hand, inhibition of DAG lipase prevents persistent eCB-dependent eIPSC suppression triggered by mAChRs. The results show that the biochemical pathways for the various eCB-dependent responses differ and might therefore be independently manipulated.


Novel anaerobic ultramicrobacteria belonging to the Verrucomicrobiales lineage of bacterial descent isolated by dilution culture from anoxic rice paddy soil.
The use of dilution culture techniques to cultivate saccharolytic bacteria present in the anoxic soil of flooded rice microcosms allowed the isolation of three new strains of bacteria, typified by their small cell sizes, with culturable numbers estimated at between 1.2 x 10(5) and 7.3 x 10(5) cells per g of dry soil. The average cell volumes of all three strains were 0.03 to 0.04 microns3, and therefore they can be termed ultramicrobacteria or "dwarf cells." The small cell size is a stable characteristic, even when the organisms grow at high substrate concentrations, and thus is not a starvation response. All three strains have genomic DNA with a mol% G+C ratio of about 63, are gram negative, and are motile by means of a single flagellum. The three new isolates utilized only sugars and some sugar polymers as substrates for growth. The metabolism is strictly fermentative, but the new strains are oxygen tolerant. Sugars are metabolized to acetate, propionate, and succinate. Hydrogen production was not significant. In the presence of 0.2 atm of oxygen, the fermentation end products or ratios did not change. The phylogenetic analysis on the basis of 16S ribosomal DNA (rDNA) sequence comparisons indicates that the new isolates belong to a branch of the Verrucomicrobiales lineage and are closely related to a cloned 16S rDNA sequence (PAD7) recovered from rice paddy field soil from Japan. The isolation of these three strains belonging to the order Verrucomicrobiales from a model rice paddy system, in which rice was grown in soil from an Italian rice field, provides some information on the possible physiology and phenotype of the organism represented by the cloned 16S rDNA sequence PAD7. The new isolates also extend our knowledge on the phenotypic and phylogenetic depths of members of the order Verrucomicrobiales, to date acquired mainly from cloned 16S rDNA sequences from soils and other habitats.


Assessing bottled water nitrate concentrations to evaluate total drinking water nitrate exposure and risk of birth defects.
Previous epidemiologic studies of maternal exposure to drinking water nitrate did not account for bottled water consumption. The objective of this National Birth Defects Prevention Study (NBDPS) (USA) analysis was to assess the impact of bottled water use on the relation between maternal exposure to drinking water nitrate and selected birth defects in infants born during 1997-2005. Prenatal residences of 1,410 mothers reporting exclusive bottled water use were geocoded and mapped; 326 bottled water samples were collected and analyzed using Environmental Protection Agency Method 300.0. Median bottled water nitrate concentrations were assigned by community; mothers' overall intake of nitrate in mg/day from drinking water was calculated. Odds ratios for neural tube defects, limb deficiencies, oral cleft defects, and heart defects were estimated using mixed-effects models for logistic regression. Odds ratios (95% CIs) for the highest exposure group in offspring of mothers reporting exclusive use of bottled water were: neural tube defects [1.42 (0.51, 3.99)], limb deficiencies [1.86 (0.51, 6.80)], oral clefts [1.43 (0.61, 3.31)], and heart defects [2.13, (0.87, 5.17)]. Bottled water nitrate had no appreciable impact on risk for birth defects in the NBDPS.


Measurement Invariance and Sleep Quality Differences Between Men and Women in the Pittsburgh Sleep Quality Index.
Previous research has reported mixed results in terms of sex differences in sleep quality. We conducted an analysis of measurement invariance of the Pittsburgh Sleep Quality Index (PSQI) between men and women to provide a necessary foundation for examining sleep differences. The sample included 861 adults (mean age = 52.73 years, 47.85% male) from the 2012-2016 wave of the Midlife in the United States (MIDUS) Refresher Biomarker survey. We randomly divided the sample into two half samples for exploratory factor analysis (EFA) and confirmatory factor analysis (CFA), respectively. We conducted EFA with a weighted least squares mean and variance adjusted (WLSMV) estimator and Geomin rotation to explore the underlying structure of the PSQI. We then employed multiple-group CFA with the WLSMV estimator and theta parameterization to examine measurement invariance between males and females. EFA suggested a two-factor structure of the PSQI, and the two-factor CFA model fit the data well. The finding that the two-factor PSQI model was invariant between males and females on configuration, factor loadings, thresholds for all but one measure, and residual variances for all but one measure provided evidence that the two-factor PSQI model was partially invariant between men and women. Females had higher means on latent factors, suggesting worse self-reports of sleep among women. Overall, the measure of the PSQI assesses the same factors in a comparable way among men and women. Women reported worse sleep than men.


SeqBuster, a bioinformatic tool for the processing and analysis of small RNAs datasets, reveals ubiquitous miRNA modifications in human embryonic cells.
High-throughput sequencing technologies enable direct approaches to catalog and analyze snapshots of the total small RNA content of living cells. Characterization of high-throughput sequencing data requires bioinformatic tools offering a wide perspective of the small RNA transcriptome. Here we present SeqBuster, a highly versatile and reliable web-based toolkit to process and analyze large-scale small RNA datasets. The high flexibility of this tool is illustrated by the multiple choices offered in the pre-analysis for mapping purposes and in the different analysis modules for data manipulation. To overcome the storage capacity limitations of the web-based tool, SeqBuster offers a stand-alone version that permits the annotation against any custom database. SeqBuster integrates multiple analyses modules in a unique platform and constitutes the first bioinformatic tool offering a deep characterization of miRNA variants (isomiRs). The application of SeqBuster to small-RNA datasets of human embryonic stem cells revealed that most miRNAs present different types of isomiRs, some of them being associated to stem cell differentiation. The exhaustive description of the isomiRs provided by SeqBuster could help to identify miRNA-variants that are relevant in physiological and pathological processes. SeqBuster is available at http://estivill_lab.crg.es/seqbuster.