Cancer and involuntary weight loss: failure to validate a prediction score.
Many patients who have involuntary weight loss have cancer. The Hernandez prediction rule includes 5 variables (elevated levels of alkaline phosphatase and lactate dehydrogenase, low albumin, high white blood cell count, and age >80 years). The purpose of this study was to evaluate the validity of the prediction rule. We prospectively evaluated 290 consecutive inpatients and outpatients who had involuntary weight loss. Clinical, hematologic, and biochemical parameters were determined. There were 259 patients who had follow-up at 6 months to determine the cause of involuntary weight loss, and 31 other patients were lost to follow-up. The 5 variables were introduced into a regression logistic model with cancer as a dependent variable. Cancer was diagnosed in 72 of the 290 patients (25%) who had involuntary weight loss. Bivariate analysis showed that serum albumin, C-reactive protein, erythrocyte sedimentation rate, alkaline phosphatase, iron, lactate dehydrogenase, white blood cell count, hemoglobin, and ferritin levels were associated with cancer (range of area under the receiver operating characteristic curve, 0.589 to 0.688). Multivariate analysis showed that albumin, erythrocyte sedimentation rate, iron, white blood cell count, and lactate dehydrogenase levels were associated with cancer. When dichotomized, only low albumin (odds ratio, 2.6, CI [1.3-5.2]) and high alkaline phosphatase (odds ratio, 2.3, CI [1.7-4.7]) were associated with cancer. The area under the receiver operating characteristic curve of the 5-variable prediction rule was only 0.70 (95% confidence interval, 0.61-0.78). The negative predictive value of this model with 3 variables (age >60 y, alkaline phosphatase, and albumin level) increased from 85% to 95% when all tests were negative. In patients who had involuntary weight loss, those who have cancer are likely to have ≥1 abnormal laboratory test. The 5-variable prediction rule had a significantly lower accuracy than originally reported. Further evaluation of the 3-variable modification of the prediction rule may be useful.


Self-replenishing surfaces.
Damaged surfaces self-replenish their chemical composition by the spontaneous re-orientation of functional groups chemically bonded to the polymer network. The repair of the surface chemistry leads to the recovery of surface functionality. This self-replenishing approach is suitable to recover many surface-related properties and constitutes a major breakthrough in extending the service life-time of functional materials.


Upregulation of Proinflammatory Bradykinin Peptides in Systemic Lupus Erythematosus and Rheumatoid Arthritis.
Our recent study has implicated bradykinin (BK) signaling as being of pathogenic importance in lupus. This study aims to investigate the biomarker potential of BK peptides, BK and BK-des-arg-9, in lupus and other rheumatic autoimmune diseases. Sera from systemic lupus erythematosus (SLE) patients and healthy subjects were screened for BK and BK-des-arg-9 by liquid chromatography-mass spectrometry metabolomics. Serum from 6-mo-old C57BL/6 mice and three murine lupus strains were also screened for the two peptides by metabolomics. Given the promising initial screening results, validation of these two peptides was next conducted using multiple reaction monitoring in larger patient cohorts. In initial metabolomics screening, BK-des-arg-9 was 22-fold higher in SLE serum and 106-fold higher in mouse lupus serum compared with healthy controls. In validation assays using multiple reaction monitoring and quadrupole time-of-flight mass spectrometry, BK and BK-des-arg-9 showed significant elevations in SLE serum compared with controls (p < 0.0001; area under the curve = 0.79-0.88), with a similar but less pronounced increase being noted in rheumatoid arthritis serum. Interestingly, increased renal SLE disease activity index in lupus patients was associated with reduced circulating BK-des-arg-9, and the reasons for this remain to be explored. To sum, increased conversion of BK to the proinflammatory metabolite BK-des-arg-9 appears to be a common theme in systemic rheumatic diseases. Besides serving as an early marker for systemic autoimmunity, independent studies also show that this metabolic axis may also be a pathogenic driver and therapeutic target in lupus.


Development of a high efficiency integration system and promoter library for rapid modification of Pseudomonas putida KT2440.
Pseudomonas putida strains are highly robust bacteria known for their ability to efficiently utilize a variety of carbon sources, including aliphatic and aromatic hydrocarbons. Recently, P. putida has been engineered to valorize the lignin stream of a lignocellulosic biomass pretreatment process. Nonetheless, when compared to platform organisms such as Escherichia coli, the toolkit for engineering P. putida is underdeveloped. Heterologous gene expression in particular is problematic. Plasmid instability and copy number variance provide challenges for replicative plasmids, while use of homologous recombination for insertion of DNA into the chromosome is slow and laborious. Further, most heterologous expression efforts to date typically rely on overexpression of exogenous pathways using a handful of poorly characterized promoters. To improve the P. putida toolkit, we developed a rapid genome integration system using the site-specific recombinase from bacteriophage Bxb1 to enable rapid, high efficiency integration of DNA into the P. putida chromosome. We also developed a library of synthetic promoters with various UP elements, -35 sequences, and -10 sequences, as well as different ribosomal binding sites. We tested these promoters using a fluorescent reporter gene, mNeonGreen, to characterize the strength of each promoter, and identified UP-element-promoter-ribosomal binding sites combinations capable of driving a ~150-fold range of protein expression levels. An additional integrating vector was developed that confers more robust kanamycin resistance when integrated at single copy into the chromosome. This genome integration and reporter systems are extensible for testing other genetic parts, such as examining terminator strength, and will allow rapid integration of heterologous pathways for metabolic engineering.


What causes EBRI retirement readiness ratings to vary: results from the 2014 Retirement Security Projection Model.
RETIREMENT INCOME ADEQUACY IMPROVED SLIGHTLY IN 2013: Due to the increase in financial market and housing values during 2013, the probability that Baby Boomers and Generation Xers would NOT run short of money in retirement increases between 0.5 and 1.6 percentage points, based on the Employee Benefit Research Institute (EBRI) Retirement Readiness Ratings (RRRs). ELIGIBILITY FOR PARTICIPATION IN AN EMPLOYER-SPONSORED DEFINED CONTRIBUTION PLAN REMAINS ONE OF THE MOST IMPORTANT FACTORS FOR RETIREMENT INCOME ADEQUACY: RRR values double for Gen Xers in the lowest-income quartile when comparing those with 20 or more years of future eligibility with those with no years of future eligibility, while those in the middle income quartiles experience increases in RRR values by 27.1-30.3 percentage points. FUTURE SOCIAL SECURITY BENEFITS MAKE A HUGE DIFFERENCE FOR THE RETIREMENT INCOME ADEQUACY OF SOME HOUSEHOLDS, ESPECIALLY GEN XERS IN THE LOWEST-INCOME QUARTILE: If Social Security benefits are subject to proportionate decreases beginning in 2033 (according to the values in Figure 8), the RRR values for those households will drop by more than 50 percent: from 20.9 percent to 10.3 percent. LONGEVITY RISK AND STOCHASTIC HEALTH CARE RISK ARE ASSOCIATED WITH HUGE VARIATIONS IN RETIREMENT INCOME ADEQUACY: For both of these factors, a comparison between the most "risky" quartile with the least risky quartile shows a spread of approximately 30 percentage points for the lowest income range, approximately 25 to 40 percentage points for the highest income range, and even larger spreads for those in the middle income ranges. A GREAT DEAL OF THE VARIABILITY IN RETIREMENT INCOME ADEQUACY COULD BE MITIGATED BY APPROPRIATE RISK-MANAGEMENT TECHNIQUES AT OR NEAR RETIREMENT AGE: For example, the annuitization of a portion of the defined contribution and IRA balances may substantially increase the probability of not running short of money in retirement. Moreover, a well-functioning market in long-term care insurance would appear to provide an extremely useful technique to help control the volatility from the stochastic, long-term health care risk, especially for those in the middle income quartiles.


Hitchhiking effects of recurrent beneficial amino acid substitutions in the Drosophila melanogaster genome.
Several recent studies have estimated that a large fraction of amino acid divergence between species of Drosophila was fixed by positive selection, using statistical approaches based on the McDonald-Kreitman test. However, little is known about associated selection coefficients of beneficial amino acid mutations. Recurrent selective sweeps associated with adaptive substitutions should leave a characteristic signature in genome variability data that contains information about the frequency and strength of selection. Here, I document a significant negative correlation between the level and the frequency of synonymous site polymorphism and the rate of protein evolution in highly recombining regions of the X chromosome of D. melanogaster. This pattern is predicted by recurrent adaptive protein evolution and suggests that adaptation is an important determinant of patterns of neutral variation genome-wide. Using a maximum likelihood approach, I estimate the product of the rate and strength of selection under a recurrent genetic hitchhiking model, lambda2N(e)s approximately 3 x 10(-8). Using an approach based on the McDonald-Kreitman test, I estimate that approximately 50% of divergent amino acids were driven to fixation by positive selection, implying that beneficial amino acid substitutions are of weak effect on average, on the order of 10(-5) (i.e., 2N(e)s approximately 40). Two implications of these results are that most adaptive substitutions will be difficult to detect in genome scans of selection and that population size (and genetic drift) may be an important determinant of the evolutionary dynamics of protein adaptation.


Changes in the bacterial community structure in stored wormbed leachate.
Organic wastes, such as cow manure, are often composted with earthworms (vermicomposting) while excess water is drained and collected. This wormbed leachate is nutrient-rich and it has been extensively used to fertilize plants. However, it is derived partially from a not yet finished compost process and could exhibit phytotoxicity or contain potentially hazardous microorganisms. The bacterial community in wormbed leachate derived from vermicomposting of cow manure was studied by pyrosequencing the 16S rRNA gene. The fresh wormbed leachate was rich in Mollicutes, particularly the genus Acholeplasma which contain phytopathogen species. The abundance of the Mollicutes decreased when the leachate was stored, while that of the Rhizobiales and the genus Pseudomonas increased. The bacterial communities changed rapidly in the leachate during storage. The changes in ammonium, nitrate and inorganic carbon content of the wormbed leachate when stored were correlated to changes in the bacterial community structure. It was found that storage of the wormbed leachate might be required before it can be applied to crops as large proportions of potentially plant pathogens were found in the fresh leachate.


Efficacies of different preparations of glucosamine for the treatment of osteoarthritis: a meta-analysis of randomised, double-blind, placebo-controlled trials.
To determine the efficacies of different preparations of glucosamine for the treatment of osteoarthritis (OA). Systematic searches of the bibliographic databases Medline, Embase, the Cochrane Central Register of Controlled Trials and the Cochrane Database of Systematic Reviews for randomised, double-blind, placebo-controlled trials (RCTs) concerning glucosamine treatment of OA. Effect size (ES) was estimated using Cohen's standardised mean difference. Consistency was evaluated via the I(2) index. Nineteen trials (3159 patients) contributed to the meta-analysis, revealing a large degree of inconsistency among the trials in terms of pain-reduction outcome: the combined ES in glucosamine sulphate (GS) trials was -0.22 [95% confidence intervals (CI) -0.48, 0.04], I(2) was 82.3%. The combined ES in glucosamine hydrochloride (GH) trials was -0.03 (95% CI -0.14, 0.08), with an absence of heterogeneity. No treatment ES was observed [-0.38 (95% CI -0.99, 0.23)] favouring GS in trials of less than 24 weeks duration and the I(2) remained high (I(2) = 88.5%). No significant treatment ES -0.09 (95% CI -0.21, 0.03) was observed in trials of more than 24 weeks duration compared with placebo, with a heterogeneity of zero. In terms of function-modifying outcomes, GS showed no significant effect on Lequesne Index reduction vs. placebo in trials of less than 24 weeks duration (ES -0.55 (95% CI -1.22, 0.11)) with a high degree of heterogeneity (I(2) = 92.9%). Pooling data from studies with durations of more than 24 weeks presented a significant combined ES of -0.36 (95% CI: -0.56, -0.17) with an absence of heterogeneity. No risk of publication bias could be detected using Egger test. GH is ineffective for pain reduction in patients with knee OA. GS may have function-modifying effects in patients with knee OA when administered for more than 6 months. However, it showed no pain-reduction benefits after 6 months of therapy.


The pathogenic intestinal spirochaete Brachyspira pilosicoli forms a diverse recombinant species demonstrating some local clustering of related strains and potential for zoonotic spread.
Brachyspira pilosicoli is an anaerobic spirochaete that can colonizes the large intestine of many host species. Infection is particularly problematic in pigs and adult poultry, causing colitis and diarrhea, but it is also known to result in clinical problems in human beings. Despite the economic importance of the spirochaete as an animal pathogen, and its potential as a zoonotic agent, it has not received extensive study. A multilocus sequence typing (MLST) method based on the scheme used for other Brachyspira species was applied to 131 B. pilosicoli isolates originating from different host species and geographical areas. A variety of phylogenetic trees were constructed and analyzed to help understand the data. The isolates were highly diverse, with 127 sequence types and 123 amino acid types being identified. Large numbers (50-112) of alleles were present at each locus, with all loci being highly polymorphic. The results of Shimodaira-Hasegawa tests identified extensive genetic recombination, although the calculated standardized index of association value (0.1568; P <0.0005) suggested the existence of some clonality. Strains from different host species and geographical origins generally were widely distributed throughout the population, although in nine of the ten cases where small clusters of related isolates occurred these were from the same geographical areas or farms/communities, and from the same species of origin. An exception to the latter was a cluster of Australian isolates originating from pigs, chickens and a human being, suggesting the likelihood of relatively recent transmission of members of this clonal group between species. The strongly recombinant population structure of B. pilosicoli contrasts to the more highly clonal population structures of the related species Brachyspira hyodysenteriae and Brachyspira intermedia, both of which are specialized enteric pathogens of pigs and poultry. The genomic plasticity of B. pilosicoli may help to explain why it has been able to adapt to colonize the large intestines of a wider range of hosts compared to other Brachyspira species. The identification of a clonal group of isolates that had been recovered from different host species, including a human being, suggests that zoonotic transmission by B. pilosicoli may occur in nature. Evidence for local transmission between the same host species also was obtained.


Altered expression of Rb and p53 in uveal melanomas following plaque radiotherapy.
To examine the expression of proteins in the Rb and p53 tumor suppressor pathways in uveal melanomas following plaque radiotherapy. Immunohistochemistry and cell culture studies. Immunohistochemistry for Rb, p16, cyclin D1, p53, HDM2, and Bcl-2 was performed on twelve eyes containing posterior uveal melanomas that were enucleated following plaque radiotherapy. Cell culture studies were performed in three cases. The irradiated eyes were enucleated for radiation complications (five cases), local tumor recurrence (three cases), and other reasons (four cases). On histopathologic examination, all cases showed evidence of tumor cell loss. However, residual tumor cells were present in all cases, including those that were clinically regressed. Residual cells from three of the clinically regressed cases were cultured and demonstrated minimal cell division, marked cell death, and extensive chromosomal damage. Strong p53 staining was observed in six cases (50%) and was significantly associated with recent radiotherapy (P = .04). Abnormal cytoplasmic staining for Rb was observed in four cases (33%). Plaque radiotherapy of uveal melanomas induces DNA damage, inhibits cell division, and promotes cell death. These changes may be due, at least in part, to induction of p53, which activates genes involved in both cell cycle arrest and apoptosis. Plaque radiotherapy can also cause alterations in the expression of Rb, but the significance of this finding will require further study.


Postfreeze reduction of locomotor endurance in the freeze-tolerant wood frog, Rana sylvatica.
Considerable study has focused on the physiological adaptations for freeze tolerance in the wood frog, Rana sylvatica, a northern species that overwinters within the frost zone, but little attention has been paid to the associated costs to organismal performance. Here we report that freezing causes transient impairment of locomotor endurance and adverse changes in exercise physiology that persist for at least 96 h. Wood frogs frozen at -2 degrees C for 36 h exhibited normal behaviors and hydro-osmotic status and near-normal metabolite (glycogen, glucose, and lactate) levels within 24 h after thawing began. However, when exercised to exhaustion on a treadmill, these frogs showed a 40% reduction in endurance as compared to sham-treated (unfrozen) controls, a reduction that persisted for at least 96 h. Previously frozen frogs exhibited higher rates of lactate accumulation during exercise than controls, suggesting that prior freezing forces greater reliance on the glycolytic pathways of energy production to support exercise. Given that this species breeds in late winter, when subzero temperatures are common, freezing may result in reduced fitness by hampering their ability to reach the pond, avoid predators, and successfully obtain mates.


DR_SEQAN: a PC/Windows-based software to evaluate drug resistance using human immunodeficiency virus type 1 genotypes.
Genotypic assays based on DNA sequencing of part or the whole reverse transcriptase (RT)- and protease (PR)-coding regions of the human immunodeficiency virus type 1 (HIV-1) genome have become part of the routine clinical management of HIV-infected individuals. However, the results are difficult to interpret due to complex interactions between mutations found in viral genes. DR_SEQAN is a tool to analyze RT and PR sequences. The program output includes a list containing all of the amino acid changes found in the query sequence in comparison with the sequence of a wild-type HIV-1 strain. Translation of codons containing nucleotide mixtures can result in potential ambiguities or heterogeneities in the amino acid sequence. The program identifies all possible combinations of 2 or 3 amino acids that derive from translation of triplets containing nucleotide mixtures. In addition, when ambiguities affect codons relevant for drug resistance, DR_SEQAN allows the user to select the appropriate mutation to be considered by the program's drug resistance interpretation algorithm. Resistance is predicted using a rule-based algorithm, whose efficiency and accuracy has been tested with a large set of drug susceptibility data. Drug resistance predictions given by DR_SEQAN were consistent with phenotypic data and coherent with predictions provided by other publicly available algorithms. In addition, the program output provides two tables showing published drug susceptibility data and references for mutations and combinations of mutations found in the analyzed sequence. These data are retrieved from an integrated relational database, implemented in Microsoft Access, which includes two sets of non-redundant core tables (one for combinations of mutations in the PR and the other for combinations in the RT). DR_SEQAN is an easy to use off-line application that provides expert advice on HIV genotypic resistance interpretation. It is coded in Visual Basic for use in PC/Windows-based platforms. The program is freely available under the General Public License. The program (including the integrated database), documentation and a sample sequence can be downloaded from http://www2.cbm.uam.es:8080/lmenendez/DR_SEQAN.zip.


Systems pathology--taking molecular pathology into a new dimension.
The wealth of morphological, histological, and molecular data from human cancers available to pathologists means that pathology is poised to become a truly quantitative systems science. By measuring morphological parameters such as tumor stage and grade, and by measuring molecular biomarkers such as hormone receptor status, pathologists have sometimes accurately predicted what will happen to a patient's tumor. While 'omic' technologies have seemingly improved prognostication and prediction, some molecular 'signatures' are not useful in clinical practice because of the failure to independently validate these approaches. Many associations between gene 'signatures' and clinical response are correlative rather than mechanistic, and such associations are poor predictors of how cellular biochemical networks will behave in perturbed, diseased cells. Using systems biology, the dynamics of reactions in cells and the behavior between cells can be integrated into models of cancer. The challenge is how to integrate multiple data from the clinic into tractable models using mathematical models and systems biology, and how to make the resultant model sufficiently robust to be of practical use. We discuss the difficulties in using mathematics to model cancer, and review some approaches that may be used to allow systems biology to be successfully applied in the clinic.


Experimental infection of bats with Geomyces destructans causes white-nose syndrome.
White-nose syndrome (WNS) has caused recent catastrophic declines among multiple species of bats in eastern North America. The disease's name derives from a visually apparent white growth of the newly discovered fungus Geomyces destructans on the skin (including the muzzle) of hibernating bats. Colonization of skin by this fungus is associated with characteristic cutaneous lesions that are the only consistent pathological finding related to WNS. However, the role of G. destructans in WNS remains controversial because evidence to implicate the fungus as the primary cause of this disease is lacking. The debate is fuelled, in part, by the assumption that fungal infections in mammals are most commonly associated with immune system dysfunction. Additionally, the recent discovery that G. destructans commonly colonizes the skin of bats of Europe, where no unusual bat mortality events have been reported, has generated further speculation that the fungus is an opportunistic pathogen and that other unidentified factors are the primary cause of WNS. Here we demonstrate that exposure of healthy little brown bats (Myotis lucifugus) to pure cultures of G. destructans causes WNS. Live G. destructans was subsequently cultured from diseased bats, successfully fulfilling established criteria for the determination of G. destructans as a primary pathogen. We also confirmed that WNS can be transmitted from infected bats to healthy bats through direct contact. Our results provide the first direct evidence that G. destructans is the causal agent of WNS and that the recent emergence of WNS in North America may represent translocation of the fungus to a region with a naive population of animals. Demonstration of causality is an instrumental step in elucidating the pathogenesis and epidemiology of WNS and in guiding management actions to preserve bat populations against the novel threat posed by this devastating infectious disease.


Detection of multiregion objects embedded in nonoverlapping noise.
The concept of a statistical filter for objects that comprise several regions is introduced. The process is optimal in the presence of nonoverlapping noise for the target and may perform independently of variations in the mean value in every region. The basic performance of the filter is described, and a comparison with other types of processing is made.


Recently formed polyploid plants diversify at lower rates.
Polyploidy, the doubling of genomic content, is a widespread feature, especially among plants, yet its macroevolutionary impacts are contentious. Traditionally, polyploidy has been considered an evolutionary dead end, whereas recent genomic studies suggest that polyploidy has been a key driver of macroevolutionary success. We examined the consequences of polyploidy on the time scale of genera across a diverse set of vascular plants, encompassing hundreds of inferred polyploidization events. Likelihood-based analyses indicate that polyploids generally exhibit lower speciation rates and higher extinction rates than diploids, providing the first quantitative corroboration of the dead-end hypothesis. The increased speciation rates of diploids can, in part, be ascribed to their capacity to speciate via polyploidy. Only particularly fit lineages of polyploids may persist to enjoy longer-term evolutionary success.


Long-term chloride concentrations in North American and European freshwater lakes.
Anthropogenic sources of chloride in a lake catchment, including road salt, fertilizer, and wastewater, can elevate the chloride concentration in freshwater lakes above background levels. Rising chloride concentrations can impact lake ecology and ecosystem services such as fisheries and the use of lakes as drinking water sources. To analyze the spatial extent and magnitude of increasing chloride concentrations in freshwater lakes, we amassed a database of 529 lakes in Europe and North America that had greater than or equal to ten years of chloride data. For each lake, we calculated climate statistics of mean annual total precipitation and mean monthly air temperatures from gridded global datasets. We also quantified land cover metrics, including road density and impervious surface, in buffer zones of 100 to 1,500 m surrounding the perimeter of each lake. This database represents the largest global collection of lake chloride data. We hope that long-term water quality measurements in areas outside Europe and North America can be added to the database as they become available in the future.


Virginity pledges among the willing: delays in first intercourse and consistency of condom use.
We examine longitudinal relationships between virginity pledging in adolescence and both sexual initiation and condom use. Prior studies have had mixed results and may not adequately control for prepledge differences between pledgers and nonpledgers. Data came from a national sample of 12- to 17-year-olds surveyed in 2001 and reinterviewed 1 and 3 years later. Logistic regression models estimated the association between making a pledge and each outcome. Selection bias was reduced through propensity-score weighting and a rich set of demographic and psychosocial covariates. Pledgers and nonpledgers differed substantially in preexisting characteristics. However, after propensity weighting and statistical controls, pledging was still associated with delayed intercourse. We estimate that in the absence of pledging 42.4% of virgins with characteristics indicating an inclination to pledge initiate intercourse within 3 years; in the presence of the pledge, 33.6% of such youth initiate intercourse. Among those who had sex during this period, pledging was unassociated with condom use. Among those who did not have sex during this period, pledging was unassociated with engagement in noncoital sexual behavior. Making a virginity pledge appears to be an effective means of delaying sexual intercourse initiation among those inclined to pledge without influencing other sexual behavior; pledging does not appear to affect sexual safety among pledgers who fail to remain abstinent.


Fate Mapping Quantifies the Dynamics of B Cell Development and Activation throughout Life.
Follicular mature (FM) and germinal center (GC) B cells underpin humoral immunity, but the dynamics of their generation and maintenance are not clearly defined. Here, we exploited a fate-mapping system in mice that tracks B cells as they develop into peripheral subsets, together with a cell division fate reporter mouse and mathematical models. We find that FM cells are kinetically homogeneous, recirculate freely, are continually replenished from transitional populations, and self-renew rarely. In contrast, GC B cell lineages persist for weeks with rapid turnover and site-specific dynamics. Those in the spleen derive from transitional cells and are kinetically homogeneous, while those in lymph nodes derive from FM B cells and comprise both transient and persistent clones. These differences likely derive from the nature of antigen exposure at the different sites. Our integrative approach also reveals how the host environment drives cell-extrinsic, age-related changes in B cell homeostasis.


Angiotensin II activates signal transducer and activators of transcription 3 via Rac1 in atrial myocytes and fibroblasts: implication for the therapeutic effect of statin in atrial structural remodeling.
Recently, activation of the local renin-angiotensin system and mitogen-activated protein kinase pathways in atrial myocardium has been found to play an important role in atrial structural remodeling related to atrial fibrillation. Another important mediator of the angiotensin II (Ang II) effect is the Janus kinase/signal transducers and activators of transcription (STAT) pathway, which has never been characterized in the atrium. In cultured atrial myocytes and fibroblasts, Ang II induced tyrosine phosphorylation of STAT3 through a Rac1-dependent mechanism, which was inhibited by dominant-negative Rac1, losartan, and simvastatin. In atrial myocytes, activation of STAT3 by Rac1 was mediated by direct association of Rac1 with STAT3; however, in atrial fibroblasts, it was mediated by an indirect paracrine effect. Constitutively active STAT3 increased protein synthesis, and dominant-negative STAT3 abrogated Ang II-induced protein synthesis in atrial myocytes and fibroblasts. Rats infused long term with Ang II exhibited higher levels of activated Rac1, phospho-STAT3, collagen synthesis, and atrial fibrosis in the atria, all of which were attenuated by oral losartan and simvastatin. In human atrial tissues from patients with atrial fibrillation, Ang II and phospho-STAT3 levels were also elevated. The Ang II/Rac1/STAT3 pathway is an important signaling pathway in the atrial myocardium to mediate atrial structural remodeling, and losartan and statin may be able to reverse Ang II-induced atrial structural remodeling in atrial fibrillation.


Greater Trochanteric Pain Syndrome.
Patients who have lateral hip pain historically have been diagnosed with trochanteric bursitis and treated with nonsteroidal anti-inflammatory medications, corticosteroid injections, and physical therapy. Although this strategy is effective for most patients, a substantial number of patients continue to have pain and functional limitations. Over the past decade, our understanding of disorders occurring in the peritrochanteric space has increased dramatically. Greater trochanteric pain syndrome encompasses trochanteric bursitis, external coxa saltans (ie, snapping hip), and abductor tendinopathy. A thorough understanding of the anatomy, examination findings, and imaging characteristics aids the clinician in treating these patients. Open and endoscopic treatment options are available for use when nonsurgical treatment is unsuccessful.


Incidence of postpartum infection after vaginal delivery in Viet Nam.
This study assessed the incidence of postpartum infection which is rarely clinically evaluated and is probably underestimated in developing countries. This prospective study identified infection after vaginal delivery by clinical and laboratory examinations prior to discharge from hospital and again at six weeks postpartum in Ho Chi Minh City, Viet Nam. Textbook definitions, physicians' diagnoses, symptomatic and verbal autopsy definitions were used for classifying infection. Logistic regression was used for determining associations of postpartum infection with socioeconomic and reproductive characteristics. In total, 978 consecutive, eligible consenting women were followed up at 42+/-7 (range 2-45) days postpartum (not associated with incidence). Ninety-eight percent took 'prophylactic' antibiotics. The most conservative estimate of the incidence of postpartum infection was 1.7%. The incidence of serious infection was 0.5%, but increased to 4.6% when verbal autopsy and symptomatic definitions were used. Postpartum infection, particularly serious infection, is greatly underestimated. Just preventing or treating infection could have a substantial impact on reducing maternal mortality in developing countries.


Theoretical analysis of two nonpolarizing beam splitters in asymmetrical glass cubes.
The design principle for a nonpolarizing beam splitter based on the Brewster condition in a cube is introduced. Nonpolarizing beam splitters in an asymmetrical glass cube are proposed and theoretically investigated, and applied examples are given. To realize 50% reflectance and 50% transmittance at specified wavelengths for both polarization components with an error of less than 2%, two measurements are taken by adjusting the refractive index of the substrate material and optimizing the thicknesses of each film in the design procedures. The simulated results show that the targets are achieved using the method reported here.


Reflectivity changes of Ge and the efficiency of moving diffraction gratings.
Reflectivity changes and the efficiency of a temporary grating formed on a Ge surface by two beams, which differ in frequency by 26 MHz, are compared with those measured for beams of equal frequency. The results indicate that the observations can be explained by a deterioration in the quality of the grating due to its motion across the surface. The predicted effects due to a phase difference between the intensity pattern and the temperature variation on the surface which it produces were not observed.


Molecular and epidemiological characterization of imported malaria cases in Chile.
Chile is one of the South American countries certified as malaria-free since 1945. However, the recent increase of imported malaria cases and the presence of the vector Anopheles pseudopunctipennis in previously endemic areas in Chile require an active malaria surveillance programme. Specimens from 268 suspected malaria cases-all imported-collected between 2015 and 2018 at the Public Health Institute of Chile (ISP), were diagnosed by microscopy and positive cases were included for epidemiological analysis. A photo-induced electron transfer fluorogenic primer real-time PCR (PET-PCR) was used to confirm the presence of malaria parasites in available blood samples. Sanger sequencing of drug resistance molecular markers (pfk13, pfcrt and pfmdr1) and microsatellite (MS) analysis were performed in confirmed Plasmodium falciparum samples and results were related to origin of infection. Out of the 268 suspected cases, 65 were Plasmodium spp. positive by microscopy. A total of 63% of the malaria patients were male and 37% were female; 43/65 of the patients acquired infections in South American endemic countries. Species confirmation of available blood samples by PET-PCR revealed that 15 samples were positive for P. falciparum, 27 for Plasmodium vivax and 4 were mixed infections. The P. falciparum samples sequenced contained four mutant pfcrt genotypes (CVMNT, CVMET, CVIET and SVMNT) and three mutant pfmdr1 genotypes (Y184F/S1034C/N1042D/D1246Y, Y184F/N1042D/D1246Y and Y184F). MS analysis confirmed that all P. falciparum samples presented different haplotypes according to the suspected country of origin. Four patients with P. vivax infection returned to the health facilities due to relapses. The timely detection of polymorphisms associated with drug resistance will contribute to understanding if current drug policies in the country are appropriate for treatment of imported malaria cases and provide information about the most frequent resistant genotypes entering Chile.


Human embryonic stem cell derived hepatocyte-like cells as a tool for in vitro hazard assessment of chemical carcinogenicity.
Hepatocyte-like cells derived from the differentiation of human embryonic stem cells (hES-Hep) have potential to provide a human relevant in vitro test system in which to evaluate the carcinogenic hazard of chemicals. In this study, we have investigated this potential using a panel of 15 chemicals classified as noncarcinogens, genotoxic carcinogens, and nongenotoxic carcinogens and measured whole-genome transcriptome responses with gene expression microarrays. We applied an ANOVA model that identified 592 genes highly discriminative for the panel of chemicals. Supervised classification with these genes achieved a cross-validation accuracy of > 95%. Moreover, the expression of the response genes in hES-Hep was strongly correlated with that in human primary hepatocytes cultured in vitro. In order to infer mechanistic information on the consequences of chemical exposure in hES-Hep, we developed a computational method that measures the responses of biochemical pathways to the panel of treatments and showed that these responses were discriminative for the three toxicity classes and linked to carcinogenesis through p53, mitogen-activated protein kinases, and apoptosis pathway modules. It could further be shown that the discrimination of toxicity classes was improved when analyzing the microarray data at the pathway level. In summary, our results demonstrate, for the first time, the potential of human embryonic stem cell--derived hepatic cells as an in vitro model for hazard assessment of chemical carcinogenesis, although it should be noted that more compounds are needed to test the robustness of the assay.


Renal function after pediatric intestinal transplant.
Data were analyzed from 44 patients who survived more than 2 years after intestinal transplantation performed between 1994 and 2002. Median age was 1.7 years. Tacrolimus level was defined as average tacrolimus level over 6 months. Kidney function was evaluated using a 6-month average serum creatinine. Glomerular filtration rate (GFR) was calculated with the Schwartz formula. The procedures were: isolated intestinal transplantation (n = 11), liver and intestinal transplantation (n = 9), multivisceral transplantation (n = 22), and modified multivisceral transplantation (n = 2). Forty-four patients were followed for a mean of 3.6 years on tacrolimus. Tacrolimus levels ranged between 3.5 and 19.9 ng/mL (median 14.6 ng/mL) at 0 to 6 months and 6.0 to 18.9 ng/mL (median 13.2 ng/mL) at 0 to 12 months. Pretransplant kidney function as mean GFR was 138 +/- 42 mL/min/1.73 m(2) (n = 44), posttransplant kidney function at 18 to 24 month as mean GFR was 102+/-35 mL/min/1.73 m(2) (n = 44), a value that was 81% of the pretransplant GFR (P < .0001). In an analysis of tacrolimus level versus renal function, a value greater than 13.5 ng/mL during the first 12 months was a significant predictor for impaired renal function at 2 years after transplantation (defined as average GFR less than 90 mL/min/1.73 m(2) at 18 to 24 months; P = .001). Only age among age, sex, diagnosis, transplant type, and rejection episodes showed a correlation with renal function. Renal function dropped significantly at 2 years after pediatric intestinal transplantation to 81% of the pretransplantation value. Tacrolimus level for the first 12 months seemed to predict subsequent development of renal impairment at 2 years.


Assessment of occupant-behavior-based indoor air quality and its impacts on human exposure risk: A case study based on the wildfires in Northern California.
The recent wildfires in California, U.S., have caused not only significant losses to human life and property, but also serious environmental and health issues. Ambient air pollution from combustion during the fires could increase indoor exposure risks to toxic gases and particles, further exacerbating respiratory conditions. This work aims at addressing existing knowledge gaps in understanding how indoor air quality is affected by outdoor air pollutants during wildfires-by taking into account occupant behaviors (e.g., movement, operation of windows and air-conditioning) which strongly influence building performance and occupant comfort. A novel modeling framework was developed to simulate the indoor exposure risks considering the impact of occupant behaviors by integrating building energy and occupant behaviour modeling with computational fluid dynamics simulation. Occupant behaviors were found to exert significant impacts on indoor air flow patterns and pollutant concentrations, based on which, certain behaviors are recommended during wildfires. Further, the actual respiratory injury level under such outdoor conditions was predicted. The modeling framework and the findings enable a deeper understanding of the actual health impacts of wildfires, as well as informing strategies for mitigating occupant health risk during wildfires.


Neural network assisted analysis of bimetallic nanocatalysts using X-ray absorption near edge structure spectroscopy.
X-ray absorption spectroscopy is a common method for probing the local structure of nanocatalysts. One portion of the X-ray absorption spectrum, the X-ray absorption near edge structure (XANES) is a useful alternative to the commonly used extended X-ray absorption fine structure (EXAFS) for probing three-dimensional geometry around each type of atomic species, especially in those cases when the EXAFS data quality is limited by harsh reaction conditions and low metal loading. A methodology for quantitative determination of bimetallic architectures from their XANES spectra is currently lacking. We have developed a method, based on the artificial neural network, trained on ab initio site-specific XANES calculations, that enables accurate and rapid reconstruction of the structural descriptors (partial coordination numbers) from the experimental XANES data. We demonstrate the utility of this method on the example of a series of PdAu bimetallic nanoalloys. By validating the neural network-yielded metal-metal coordination numbers based on the XANES analysis by previous EXAFS characterization, we obtained new results for in situ restructuring of dilute (2.6 at% Pd in Au) PdAu nanoparticles, driven by their gas and temperature treatments.


Alteration of life span of mice chronically exposed to 2.45 GHz CW microwaves.
Female CD1 mice were exposed from the thirty-fifth day of age for the remainder of their lives to 2.45 GHz, CW-microwave radiation at a power density of 3 or 10 mW/cm2 (SAR = 2.0 or 6.8 W/kg). Exposures took place 1 h/day, 5 day/week in an anechoic chamber at an ambient temperature of 22 degrees C and a relative humidity of 50%. There were 25 animals in each exposure group, and an equal number of controls were concurrently sham exposed. The average life span of animals exposed at 10 mW/cm2 was significantly shorter than that of sham-exposed controls (572 days vs. 706 days; P = .049; truncation > 20%). In contrast, the average lifespan of the animals exposed at 3 mW/cm2 was slightly, but not significantly, longer (738 days) than that of controls (706 days).


Effect of protease inhibitors on steady-state pharmacokinetics of oral norethindrone contraception in HIV-infected women.
Pharmacokinetic interactions exist between combined oral contraceptives and protease inhibitors (PI). However, such information is lacking for progestin-only oral contraception. We sought to define the steady-state pharmacokinetic interaction between norethindrone (NET) and PI in HIV-infected women. We conducted an open-label, prospective, nonrandomized trial to characterize the steady-state pharmacokinetics of serum NET in HIV-infected women receiving PI compared with a control group of HIV-infected women receiving other noninteracting drugs. After 21 days of 0.35 mg of NET ingestion once daily, serial serum samples were obtained at 0, 1, 2, 3, 4, 6, 8, 12, 24, 48, and 72 hours. The area under the curve between 0 and 72 hours after ingestion was calculated by trapezoidal approximation. Thirty-five women were enrolled, 2 withdrew. Sixteen women in the PI group and 17 controls completed the study. NET half-life and maximum concentration were not significantly different between the 2 groups. Minimum concentration of NET was significantly higher in the PI group (P = 0.01). The ratio of the geometric mean NET area under the curve in the PI group compared with controls was 1.5 (90% confidence interval: 1.21 to 1.86). NET serum concentrations were significantly higher in HIV-infected women taking a PI compared with controls (P = 0.004). Coadministration of PI inhibits NET metabolism as shown by higher serum NET area under the curve levels, a surrogate marker for therapeutic contraceptive efficacy. This study supports the increased utilization of progestin-only pills in HIV-infected women receiving certain PI regimens.


Erythrocyte membrane calcium adenosine 5'-triphosphatase activity in the spontaneously hypertensive rat.
1. Ca2+-adenosine 5'-triphosphatase (Ca2+-Mg2+-ATPase) activity was studied simultaneously in calmodulin-deficient erythrocyte ghost membranes and inside-out vesicles (IOVs) from 12-week-old female spontaneously hypertensive rats (SHR) and their matched controls: [Wistar-Kyoto normotensive rats (WKY)], and in detergent extracts of ghost membranes. 2. Both adenosine 5'-triphosphate (ATP)-dependent Ca2+ uptake by IOVs and Ca2+-dependent ATP hydrolysis activity of ghost membranes were reduced significantly in the SHR compared with WKY, when either the calmodulin-independent or calmodulin-stimulated activities were compared. 3. The ratios between Ca2+ uptake and ATP hydrolysis activities in the SHR remained approximately 1.0, showing a proportional reduction in both activities. 4. No difference in affinity for calmodulin was observed between SHR and WKY. 5. No significant difference in Ca2+-dependent ATP hydrolysis activity was observed between SHR and WKY after detergent solubilization of erythrocyte ghost membranes. 6. These results suggest that the number of Ca2+-Mg2+-ATPase units are similar in SHR and WKY and that the reduced activity in the intact SHR membrane is due to altered membrane environment.


Difficulty Disengaging Attention from Social Threat in Social Anxiety.
Selective attention to threat is believed to maintain social anxiety, yet the nature of attentional processing remains unclear. It has been posited that difficulty disengaging from threat cues may be implicated. The present study tested this hypothesis using an eye tracking paradigm to directly examine eye fixations in a non-clinical sample (N = 46). Eye movements were tracked during presentation of social cues (happy or disgust faces) embedded with non-social cues matched on dimensions of valence, threat, and arousal. Stimuli were presented for 2,000 ms to allow for examination of attention over time. Results suggest that individuals with higher social anxiety may demonstrate relative difficulty disengaging from negative social cues (i.e., disgust faces). Social anxiety was unrelated to eye movements concerning happy faces. Implications for the maintenance and etiology of social anxiety are discussed.


The impact of aminoglycosides on the dynamics of translation elongation.
Inferring antibiotic mechanisms on translation through static structures has been challenging, as biological systems are highly dynamic. Dynamic single-molecule methods are also limited to few simultaneously measurable parameters. We have circumvented these limitations with a multifaceted approach to investigate three structurally distinct aminoglycosides that bind to the aminoacyl-transfer RNA site (A site) in the prokaryotic 30S ribosomal subunit: apramycin, paromomycin, and gentamicin. Using several single-molecule fluorescence measurements combined with structural and biochemical techniques, we observed distinct changes to translational dynamics for each aminoglycoside. While all three drugs effectively inhibit translation elongation, their actions are structurally and mechanistically distinct. Apramycin does not displace A1492 and A1493 at the decoding center, as demonstrated by a solution nuclear magnetic resonance structure, causing only limited miscoding; instead, it primarily blocks translocation. Paromomycin and gentamicin, which displace A1492 and A1493, cause significant miscoding, block intersubunit rotation, and inhibit translocation. Our results show the power of combined dynamics, structural, and biochemical approaches to elucidate the complex mechanisms underlying translation and its inhibition.


Separable routes to human memory formation: dissociating task and material contributions in the prefrontal cortex.
The present study used fMRI to investigate functional dissociations across frontal regions during incidental memory formation. Subjects were imaged while encoding materials with differential access to phonological codes (nonfamous faces and nameable famous faces) under task conditions that encouraged elaborate (deep) or superficial (shallow) encoding strategies. Results revealed a functional dissociation between dorsal posterior regions of the prefrontal cortex (BA 6/44) that were sensitive to material type (famous vs. nonfamous), irrespective of the encoding task, and ventral anterior regions of the prefrontal cortex (BA 45/47) that were uniquely sensitive to task demands (deep vs. shallow), regardless of material type. Further, subjects realized a memorial advantage to the extent that they recruited these dissociable frontal regions. These results demonstrate a posterior/anterior dichotomy in the frontal cortex that underlies separable code-based routes to human memory formation.


Whole blood genome-wide gene expression profile in males after prolonged wakefulness and sleep recovery.
Although the specific functions of sleep have not been completely elucidated, the literature has suggested that sleep is essential for proper homeostasis. Sleep loss is associated with changes in behavioral, neurochemical, cellular, and metabolic function as well as impaired immune response. Using high-resolution microarrays we evaluated the gene expression profiles of healthy male volunteers who underwent 60 h of prolonged wakefulness (PW) followed by 12 h of sleep recovery (SR). Peripheral whole blood was collected at 8 am in the morning before the initiation of PW (Baseline), after the second night of PW, and one night after SR. We identified over 500 genes that were differentially expressed. Notably, these genes were related to DNA damage and repair and stress response, as well as diverse immune system responses, such as natural killer pathways including killer cell lectin-like receptors family, as well as granzymes and T-cell receptors, which play important roles in host defense. These results support the idea that sleep loss can lead to alterations in molecular processes that result in perturbation of cellular immunity, induction of inflammatory responses, and homeostatic imbalance. Moreover, expression of multiple genes was downregulated following PW and upregulated after SR compared with PW, suggesting an attempt of the body to re-establish internal homeostasis. In silico validation of alterations in the expression of CETN3, DNAJC, and CEACAM genes confirmed previous findings related to the molecular effects of sleep deprivation. Thus, the present findings confirm that the effects of sleep loss are not restricted to the brain and can occur intensely in peripheral tissues.


Modelling the efficiency of local versus central provision of intravenous thrombolysis after acute ischemic stroke.
Prehospital redirection of stroke patients to a regional center is used as a strategy to maximize the provision of intravenous thrombolysis. We developed a model to quantify the benefit of redirection away from local services that were already providing thrombolysis. A microsimulation using hospital and ambulance data from consecutive emergency admissions to 10 local acute stroke units estimated the effect of redirection to 2 regional neuroscience centers. Modeled outcomes reflected additional journey time and accuracy of stroke identification in the prehospital phase, and the relative efficiency of patient selection and door-needle time for each local site compared with the nearest regional neuroscience center. Thrombolysis was received by 223/1884 emergency admissions. Based on observed site performance, 68 additional patients would have been treated after theoretical redirection of 1269 true positive cases and 363 stroke mimics to the neuroscience center. Over 5 years redirection of this cohort generated 12.6 quality-adjusted life years at a marginal cost of £6730 ($10,320, €8347). The average additional cost of a quality-adjusted life year gain was £534 ($819, €673). Under these specific circumstances, redirection would have improved outcomes from thrombolysis at little additional cost.


Quantification of the completeness of follow-up.
Completeness of follow-up is important, especially in clinical trials, since unequal follow-up in the treatment groups can bias the analysis of results. In survival studies, information on participants who do not complete the study is often omitted because their data can be included up to the time at which they were lost to follow-up. We propose a simple measure of completeness that is the ratio of the total observed person-time and the potential person-time of follow-up in a study. Our measure is easy to calculate, can be illustrated pictorially, and can be used to identify subgroups with especially poor follow-up.


POCS-based restoration of space-varying blurred images.
We propose a new method for space-varying image restoration using the method of projection onto convex sets (POCS). The formulation allows the use of a different blurring function at each pixel of the image in a computationally efficient manner. We illustrate the performance of the proposed approach by comparing the new results with those of the ROMKF method on simulated images. We also present results on a real-life image with unknown space-varying out-of-focus blur.


Vertebral fracture assessment (VFA) in patients over 50 years of age with a non-severe peripheral fracture.
The prevalence of unknown vertebral fractures evaluated by systematic vertebral fracture assessment (VFA) was 21% in patients over 50 years of age who suffered from a recent low-trauma non-severe peripheral fracture. The outcome of VFA resulted in changes in the management of osteoporosis. The aim of this study was to evaluate the utility of VFA in detecting vertebral fractures (VFs) in patients over 50 years of age, who suffered from a recent low-trauma non-severe peripheral fracture. This was an observational, single-center, cross-sectional study conducted in patients over 50 years of age, who presented a recent low-trauma non-severe peripheral fracture and were identified by the Fracture Liaison Service (FLS) of Amiens University Hospital between December 2017 and March 2019. VFA was interpreted by two trained rheumatologists providing a consensual reading using Genant semi-quantitative assessment. Of the 359 eligible patients, 114 patients (31.8%) were included (mean age 65.6 ± 8.4 years; 89.5% female). Twenty-four patients (21%) had one or more VF diagnosed by VFA. The total number of VF diagnosed by VFA was 30: 20 VF (66.7%) grade 1, 7 VF (23.3%) grade 2, and 3 VF (10%) grade 3. Among the 24 patients with at least one prevalent VF diagnosed by VFA, 18 patients had an osteoporosis medication adaptation after the VFA results (16 osteoporosis medication initiation and 2 treatment intensification), and 6 patients would have had an osteoporosis medication even without the VFA results (66.7% versus 33.3% respectively, p < 0.001). Of the 51 patients receiving an osteoporosis medication after DXA and VFA, 18 patients (35.3%) had a change in the management of osteoporosis after knowing the outcome of VFA. All the VFs diagnosed by VFA were unknown before. We did not evidence any threshold (age, T-score, height loss) below which no VF was detected. Our study demonstrates the usefulness of systematic VFA to detect prevalent VF in patients over 50 years of age who suffer from a recent non-severe peripheral fracture.


Construction of physical maps for the sex-specific regions of papaya sex chromosomes.
Papaya is a major fruit crop in tropical and subtropical regions worldwide. It is trioecious with three sex forms: male, female, and hermaphrodite. Sex determination is controlled by a pair of nascent sex chromosomes with two slightly different Y chromosomes, Y for male and Yh for hermaphrodite. The sex chromosome genotypes are XY (male), XYh (hermaphrodite), and XX (female). The papaya hermaphrodite-specific Yh chromosome region (HSY) is pericentromeric and heterochromatic. Physical mapping of HSY and its X counterpart is essential for sequencing these regions and uncovering the early events of sex chromosome evolution and to identify the sex determination genes for crop improvement. A reiterate chromosome walking strategy was applied to construct the two physical maps with three bacterial artificial chromosome (BAC) libraries. The HSY physical map consists of 68 overlapped BACs on the minimum tiling path, and covers all four HSY-specific Knobs. One gap remained in the region of Knob 1, the only knob structure shared between HSY and X, due to the lack of HSY-specific sequences. This gap was filled on the physical map of the HSY corresponding region in the X chromosome. The X physical map consists of 44 BACs on the minimum tiling path with one gap remaining in the middle, due to the nature of highly repetitive sequences. This gap was filled on the HSY physical map. The borders of the non-recombining HSY were defined genetically by fine mapping using 1460 F2 individuals. The genetically defined HSY spanned approximately 8.5 Mb, whereas its X counterpart extended about 5.4 Mb including a 900 Kb region containing the Knob 1 shared by the HSY and X. The 8.5 Mb HSY corresponds to 4.5 Mb of its X counterpart, showing 4 Mb (89%) DNA sequence expansion. The 89% increase of DNA sequence in HSY indicates rapid expansion of the Yh chromosome after genetic recombination was suppressed 2-3 million years ago. The genetically defined borders coincide with the common BACs on the minimum tiling paths of HSY and X. The minimum tiling paths of HSY and its X counterpart are being used for sequencing these X and Yh-specific regions.


Clinical Epidemiology (CE) and Evidence-Based Medicine (EBM) in the Asia Pacific region (Round Table Forum).
Clinical Epidemiology (CE) and Evidence-Based Medicine (EBM) have become increasingly important in an era of rising costs, patient safety concerns and evidence-based health care. CE and EBM research in the Asia Pacific region have grown significantly. However, there are three main challenges such as linking evidence to practice and policy; developing a strong collaborative network; and a need for resources and technical expertise to produce evidence. The Cochrane Collaboration is a possible solution to resolve above challenges identified, particularly the challenge of transforming evidence to practice. In addition, training can be carried out to enhance technical expertise in the region and there is also the promising potential that collaborations could extend beyond systematic reviews. To improve the adoption of evidence-based health policy, selection of the best evidence for the right audience and focusing on the relevant issues through appropriate methodology are essential. Information on effectiveness and cost effectiveness needs to be highlighted for policy makers. The way forward to strengthen research and capacity building is to establish the Asia Pacific Consortium for CE and EBM. The consortium would help to create mutually rewarding scientific research and collaborations that will augur well for advances in CE and EBM.


Evaluation of combinatorial cis-regulatory elements for stable gene expression in chicken cells.
Recent successes in biotechnological application of birds are based on their unique physiological traits such as unlimited manipulability onto developing embryos and simple protein constituents of the eggs. However it is not likely that target protein is produced as kinetically expected because various factors affect target gene expression. Although there have been various attempts to minimize the silencing of transgenes, a generalized study that uses multiple cis-acting elements in chicken has not been made. The aim of the present study was to analyze whether various cis-acting elements can help to sustain transgene expression in chicken fibroblasts. We investigated the optimal transcriptional regulatory elements for enhancing stable transgene expression in chicken cells. We generated eight constructs that encode enhanced green fluorescent protein (eGFP) driven by either CMV or CAG promoters (including the control), containing three types of key regulatory elements: a chicken lysozyme matrix attachment region (cMAR), 5'-DNase I-hypersensitive sites 4 (cHS4), and the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Then we transformed immortalized chicken embryonic fibroblasts with these constructs by electroporation, and after cells were expanded under G418 selection, analyzed mRNA levels and mean fluorescence intensity (MFI) by quantitative real-time PCR and flow cytometry, respectively. We found that the copy number of each construct significantly decreased as the size of the construct increased (R2 = 0.701). A significant model effect was found in the expression level among various constructs in both mRNA and protein (P < 0.0001). Transcription with the CAG promoter was 1.6-fold higher than the CMV promoter (P = 0.027) and the level of eGFP expression activity in cMAR- or cHS4-flanked constructs increased by two- to three-fold compared to the control CMV or CAG promoter constructs. In addition, flow cytometry analysis showed that constructs having cis-acting elements decreased the level of gene silencing as well as the coefficient of variance of eGFP-expressing cells (P < 0.0001). Our current data show that an optimal combination of cis-acting elements and promoters/enhancers for sustaining gene expression in chicken cells is suggested. These results provide important information for avian transgenesis and gene function studies in poultry.


Molecularly imprinted polymer for selective extraction of 3-methylflavone-8-carboxylic acid from human urine followed by its determination using zwitterionic hydrophilic interaction liquid chromatography.
A novel water-compatible molecularly imprinted SPE combined with zwitterionic hydrophilic interaction liquid chromatography method for selective extraction and determination of 3-methylflavone-8-carboxylic acid (MFA), the main active metabolite of flavoxate in human urine, was developed and validated. The effects of progenic solvents, pH, cross linker and amount of monomer were studied to optimize the efficiency and selectivity. The molecularly imprinted polymer showed good specific adsorption capacity with an optimum of 200 μmol/g at pH 7.5 and selective extraction of MFA from human urine. The recovery of MFA from human urine was >98%. The lower limit of quantification was 1.20 μg/mL. The proposed method overcomes the matrix effects of endogenous substances generally encountered during direct analysis of urine sample.


A Longitudinal Nine-Year Study of the Molecular Epidemiology of Carbapenemase-Producing Enterobacterales Isolated From a Regional Hospital in Taiwan: Predominance of Carbapenemase KPC-2 and OXA-48.
Enterobacterales clinical isolates are now being resistant to clinically achievable concentrations of most commonly used antibiotics that makes treatment of hospitalized patients very challenging. We hereby determine the molecular characteristics of carbapenemase genes in carbapenem-resistant Enterobacterales (CRE) isolates in Taiwan. A total of 455 CRE isolates were identified between August 2011 to July 2020. Minimum inhibitory concentrations for selected carbapenems were tested using Vitek 2, and carbapenemase genes were determined using polymerase chain reaction in combination with sequencing. Phenotypic detection of carbapenemase was determined by modified carbapenem inactivation method (mCIM) and EDTA-modified carbapenem inactivation method (eCIM) to validate our PCR screening results. Pulsed-field gel electrophoresis (PFGE) was used to determine the clonality of carbapenemase-producing Enterobacterales (CPE) isolates, and the transferability of carbapenemase-carrying plasmids was determined by conjugation assays. A slight increase in carbapenem-resistant E. coli (CREC) was observed, however, the prevalence of carbapenem-resistant K. pneumoniae (CRKP) was steady, during 2011-2020. The dominant species among our CRE was K. pneumoniae (270/455, 59.3%), followed by E. coli (81/455, 17.8%), Morganella morganii (32/455, 7.0%), and Enterobacter cloacae (25/455, 5.5%). From 2011 to 2020, the total percentage of CPE increased steadily, accounting for 61.0% of CRE in 2020. Moreover, 122 of 455 CRE isolates (26.8%) were CPE. Among the CPE isolates, the dominant carbapenemase gene was bla OXA-48-like (54/122, 44.3%), and the second most common carbapenemase gene was bla KPC-2 (47/122, 38.5%). The sensitivity and specificity for mCIM to detect carbapenemase in the 455 isolates were both 100% in this study. The PFGE results showed that 39 carbapenemase-producing E. coli and 69 carbapenemase-producing K. pneumoniae isolates carrying bla KPC-2 and/or bla NDM-5 could be classified into 5 and 12 clusters, respectively. In conclusion, our results showed an increase in CPE isolates in Taiwan. Moreover, the distribution of carbapenemase and antimicrobial susceptibility in CPE were associated with PFGE typing.


A definition and classification of status epilepticus--Report of the ILAE Task Force on Classification of Status Epilepticus.
The Commission on Classification and Terminology and the Commission on Epidemiology of the International League Against Epilepsy (ILAE) have charged a Task Force to revise concepts, definition, and classification of status epilepticus (SE). The proposed new definition of SE is as follows: Status epilepticus is a condition resulting either from the failure of the mechanisms responsible for seizure termination or from the initiation of mechanisms, which lead to abnormally, prolonged seizures (after time point t1 ). It is a condition, which can have long-term consequences (after time point t2 ), including neuronal death, neuronal injury, and alteration of neuronal networks, depending on the type and duration of seizures. This definition is conceptual, with two operational dimensions: the first is the length of the seizure and the time point (t1 ) beyond which the seizure should be regarded as "continuous seizure activity." The second time point (t2 ) is the time of ongoing seizure activity after which there is a risk of long-term consequences. In the case of convulsive (tonic-clonic) SE, both time points (t1 at 5 min and t2 at 30 min) are based on animal experiments and clinical research. This evidence is incomplete, and there is furthermore considerable variation, so these time points should be considered as the best estimates currently available. Data are not yet available for other forms of SE, but as knowledge and understanding increase, time points can be defined for specific forms of SE based on scientific evidence and incorporated into the definition, without changing the underlying concepts. A new diagnostic classification system of SE is proposed, which will provide a framework for clinical diagnosis, investigation, and therapeutic approaches for each patient. There are four axes: (1) semiology; (2) etiology; (3) electroencephalography (EEG) correlates; and (4) age. Axis 1 (semiology) lists different forms of SE divided into those with prominent motor systems, those without prominent motor systems, and currently indeterminate conditions (such as acute confusional states with epileptiform EEG patterns). Axis 2 (etiology) is divided into subcategories of known and unknown causes. Axis 3 (EEG correlates) adopts the latest recommendations by consensus panels to use the following descriptors for the EEG: name of pattern, morphology, location, time-related features, modulation, and effect of intervention. Finally, axis 4 divides age groups into neonatal, infancy, childhood, adolescent and adulthood, and elderly.


Different Populations Agree on Which Moral Arguments Underlie Which Opinions.
People often justify their moral opinions by referring to larger moral concerns (e. g., "It is unfair if homosexuals are not allowed to marry!" vs. "Letting homosexuals marry is against our traditions!"). Is there a general agreement about what concerns apply to different moral opinions? We used surveys in the United States and the United Kingdom to measure the perceived applicability of eight concerns (harm, violence, fairness, liberty, authority, ingroup, purity, and governmental overreach) to a wide range of moral opinions. Within countries, argument applicability scores were largely similar whether they were calculated among women or men, among young or old, among liberals or conservatives, or among people with or without higher education. Thus, the applicability of a given moral concern to a specific opinion can be viewed as an objective quality of the opinion, largely independent of the population in which it is measured. Finally, we used similar surveys in Israel and Brazil to establish that this independence of populations also extended to populations in different countries. However, the extent to which this holds across cultures beyond those included in the current study is still an open question.


Identification of a novel herpes simplex virus type 1-induced glycoprotein which complexes with gE and binds immunoglobulin.
We detected a glycoprotein on the surface of cells infected with herpes simplex virus type 1 (HSV-1) which, in conjunction with gE, binds immunoglobulin G (IgG). The novel glycoprotein, which has an apparent molecular mass of 70 kilodaltons and was provisionally named g70, was first detected in extracts of HSV-1-infected cells labeled by lactoperoxidase-catalyzed iodination and precipitated with rabbit sera or IgG and protein A-Sepharose. In subsequent experiments, g70 and gE were coprecipitated from extracts of HSV-1-infected cells labeled with [35S]methionine, [35S]cysteine, or 14C-amino acids. We were unable to precipitate a polypeptide analogous to g70 or gE from extracts of HSV-2-infected cells with rabbit IgG and protein A-Sepharose. Partial proteolytic peptide analysis indicated that g70 is structurally distinct from gE and gI). In addition, g70 was electrophoretically distinct from the HSV-1 Us4 glycoprotein gG. HSV-1 gE, expressed in mouse cells transfected with the gE gene, was not precipitated with rabbit IgG, nor could these cells bind radiolabeled IgG, suggesting that gE alone cannot act as an IgG (Fc) receptor. This result, coupled with the findings that gE and g70 are coprecipitated with IgG and with an anti-gE monoclonal antibody, suggests that gE and g70 form a complex which binds IgG. The electrophoretic mobilities of g70 molecules induced by different strains of HSV-1 differed markedly, arguing that g70 is encoded by the virus and is not a cellular protein induced by virus infection.


Purinergic receptor activation inhibits mitogen-stimulated proliferation in primary neurospheres from the adult mouse subventricular zone.
The expression pattern of purinergic receptors was examined in subventricular zone-derived primary neurospheres. Primary neurospheres expressed mRNA for P2X4 and P2X7 receptors, all P2Y receptors, with the exception of P2Y4, and the A1, A2a and A2b adenosine receptors. ATPgammaS, ADPbetaS and UTP evoked transient increases in cytoplasmic Ca(2+) concentration in dissociated primary neurospheres, demonstrating the functional expression of P2Y1 and P2Y2 receptors. Ca(2+) transients were not attenuated by the removal of extracellular Ca(2+) and were reversibly inhibited by the P2Y1 selective antagonist, MRS 2179. P2Y and adenosine receptor agonists reduced the size and frequency of primary neurospheres. The effects of ADPbetaS and adenosine were reversed by subtype-selective receptor antagonists, demonstrating that P2Y1 and A2a receptors mediate inhibitory effects on primary neurosphere proliferation. The modulation of neural precursor cell proliferation by P2Y and adenosine receptors therefore represents a potential regulatory mechanism within the neurogenic microenvironment.


Results at age 8 years of early intervention for low-birth-weight premature infants. The Infant Health and Development Program.
To reevaluate at age 8 years children who had participated during the first 3 years of life in a randomized clinical trial of special services for low-birthweight (LBW) premature infants. Follow-up of a randomized controlled trial of premature infants (< or = 37 weeks' gestation), stratified by 2 LBW groups (lighter [< or = 2000 g] and heavier [2001-2500 g]) and divided into intervention (n=377) and follow-up only (n=608) groups. Eight sites serving diverse populations. At age 8 years, 874 children were assessed: 336 in the intervention group and 538 in the follow-up only group. The 3-year intervention consisted of home visits (birth to 3 years), child development center services (ages 1 to 3 years), and parent group meetings (ages 1 to 3 years). Cognitive functioning (Weschler Intelligence Scale for Children-III; Peabody Picture Vocabulary Test-Revised); academic achievement (Woodcock-Johnson Tests of Achievement-Revised); and parental reports of school performance, behavior (Child Behavior Checklist), and health (Child General Health Survey). At age 8 years, in the entire cohort and in the lighter LBW stratum, the intervention and follow-up only groups were similar on all primary outcome measures. Differences favoring the intervention group were found within the heavier LBW group: full-scale IQ score (4.4 points higher, P=.007), verbal IQ score (4.2 points higher, P=.01), performance IQ score (3.9 points higher, P=.02), mathematics achievement score (4.8 points higher, P=.04), and receptive vocabulary score (6.7 points higher, P=.001). On a physical functioning subscale, the whole intervention group received less favorable ratings, while the lighter LBW intervention group had lower maternal ratings assessing social limitations caused by behavior. Although at age 8 years there were modest intervention-related differences in the cognitive and academic skills of heavier LBW premature children, attenuation of the large favorable effects seen at 3 years was observed in both the heavier and lighter LBW groups. This indicates a need to develop additional intervention strategies for LBW premature children that can provide sustained benefits.


Hypothermia Identifies Dynamin as a Potential Therapeutic Target in Experimental Stroke.
Apoptosis is a cell death pathway that is activated in ischemic stroke. The interaction between Fas and its ligand (FasL) initiates a complex pattern of intracellular events involving the recruitment of specific adaptor proteins and the development of apoptosis. We recently reported that dynamin is increased after experimental stroke, and its inhibition improves neurological outcome. Dynamin has been shown to transport Fas from the endoplasmic reticulum to the cell surface where it can be bound by its ligand, FasL. Hypothermia has been shown to improve outcome in numerous stroke models, and this protection is associated with reduced apoptosis and Fas expression. To explore the contribution of dynamin to hypothermic neuroprotection, we subjected mice to distal middle cerebral artery occlusion (dMCAO) and applied one of two cooling paradigms: one where cooling began at the onset of dMCAO (early hypothermia) and another where cooling began 1 hour later (delayed hypothermia), compared with normothermia (Norm). Both cooling paradigms reduced numbers of apoptotic cells, as well as Fas and dynamin compared with Norm. Fas and dynamin were co-expressed in neurons. Neuronal cultures were exposed to oxygen glucose deprivation. Hypothermia decreased dynamin as well as surface expression of Fas, and this correlated to reduced cell death. The results of this study suggest that dynamin may participate in the Fas-mediated apoptotic pathway, and its reduction may be linked to hypothermic neuroprotection.


Cluster analyses of grid-pattern display in macular parameters using optical coherence tomography for glaucoma diagnosis.
Using spectral-domain optical coherence tomography (SD-OCT), we assessed the ability of cluster analyses, based on the grid-pattern of macular parameters, to detect glaucoma. We enrolled 75 normal eyes, 64 early glaucomatous eyes (EG), and 40 preperimetric glaucomatous eyes (PPG). Each participant was imaged using 3-dimensional optical coherence tomography (3D-OCT) to examine the macular retinal nerve fiber layer (mRNFL) and the thickness of the ganglion cell layer, together with the inner plexiform layer (GCL/IPL). Diagnostic criteria based on the clustering of abnormal grids from the mRNFL and GCL/IPL measurements were applied. The sensitivity and specificity of glaucoma detection were compared between the cluster criteria (CC) and the average thickness criteria (ATC) of total and hemiretinal sectors, and the cut-off criteria were determined using receiver operating characteristic (ROC) curve analyses from our normal controls. The specificity values of CC and ATC from mRNFL measurements were 97% and 100%, respectively. The sensitivity of CC was 94% for EG and 68% for PPG. The sensitivity of ATC was 81% for EG and 38% for PPG. The specificity values of CC and ATC from GCL/IPL measurements were 96% and 100%, respectively. The sensitivity values of CC and ATC were 92% for EG and 63% for PPG. The sensitivity of ATC was 84% for EG and 25% for PPG. When compared to ATC and ROC-based cut-off criteria, CC showed a higher diagnostic capability. Judging abnormality based on a clustering of abnormal grids from macular OCT parameters may be a reliable approach for diagnosing early glaucoma. (http://www.umin.ac.jp/ctr/index/htm9 number, UMIN000006900.).


Intravenous fluids for migraine: a post hoc analysis of clinical trial data.
A total of 1.2 million patients present to US emergency departments (EDs) annually with migraine headache. Intravenous fluid (IVF) hydration is used to treat acute migraine commonly. We were unable to identify published data to support or refute this practice. The goal of this analysis was to determine if administration of IVF is associated with improved short-term (1 hour) or sustained (24 hours) migraine outcomes. This was a post hoc analysis of data collected from 4 ED-based migraine clinical trials in which patients were randomized to treatment with intravenous metoclopramide. In each of these studies, patients were administered IVF at the discretion of the treating physician. Our primary short-term outcome was improvement in 0 to 10 pain scale between baseline and 1 hour later. Our primary sustained outcome was the attainment of sustained headache freedom, defined as achieving a headache level of "none" in the ED and maintaining a level of "none" without headache recurrence throughout the 24- to 48-hour follow-up period. We compared mean improvement in pain scores between baseline and 1 hour later between those patients who received IVF and those who did not. We also compared the frequency of sustained headache freedom between both groups. We then used regression models to elucidate how nausea at baseline and the baseline pain score modified the relationship between IVF and the 2 outcomes. A total of 570 patients were included in the analysis. Of these, 112 (20%) were treated with IVF. Patients who received IVF improved by 4.5 (95% confidence interval [CI], 4.0-5.0) on the 0 to 10 scale, whereas patients who did not receive IVF improved by 5.1 (95% CI, 4.8-5.3) (95% CI for difference of 0.6, 0-1.1). Of patients who received IVF, 14% (95% CI, 9-22%) enjoyed sustained headache freedom vs 18% (95% CI, 15%-22%) of patients who did not (95% CI for difference of 4%, -4% to 11%). In the linear regression model, IVF was associated with less improvement in 0 to 10 pain score between baseline and 1 hour (B coefficient, -0.6; 95% CI, -1.1 to 0; P=.05). In the logistic regression model, IVF administration was not associated with sustained headache freedom (odds ratio, 0.8; 95% CI, 0.4-1.5; P=.52). Intravenous fluid did not improve pain outcomes among patients with acute migraine who were treated with intravenous metoclopramide.


The Montreal Cognitive Assessment (MoCA) is Sensitive to Head Injury and Cognitive Impairment in a Residential Alcohol and Other Drug Therapeutic Community.
Retaining clients in residential alcohol and other drug (AOD) treatment is difficult and cognitive impairment has been identified as a significant predictor of treatment dropout. The application of extensive screening for cognitive impairment is cost-prohibitive for most AOD treatment services. The current study aimed to explore cognitive functioning and impairment-associated factors in a typical sample of residential AOD clients using a free brief screening tool that could be utilised by front-line AOD services. Residents of an AOD therapeutic community (n=128) and a non-substance using control group (n=37) were administered a brief cognitive screening measure, the Montreal Cognitive Assessment (MoCA). MoCA total and domain scores were compared between these groups and within the AOD group examined in association with primary substance of misuse, severity of dependence, gender, psychological distress, and history of head injury. Almost half (43.8%) of the AOD sample were identified as cognitively impaired, compared to 16.2% of the control group. Furthermore, 67.2% of the AOD sample had sustained head injuries and 50% of the sample required hospitalization for head injury. History of head injury was a significant determinant of cognitive impairment, and associated with greater levels of psychological distress. There are high rates of inter-related cognitive impairment, head injuries, and psychological distress among clients in residential AOD treatment. Routine screening of clients at intake for cognitive impairment by means of a brief screening measure such as the MoCA, in combination with the assessment of history of head injuries and comorbid psychological disorders, could inform treatment modifications or adjunct interventions to increase retention and improve long-term outcomes.


[Hydroxyurea (hydroxycarbamide)-induced hepatic dysfunction confirmed by drug-induced lymphocyte stimulation test].
A 62-year-old man with refractory leukemia transformed from myelodysplastic syndrome was placed on hydroxyurea (hydroxycarbamide) at a daily dose of 500 mg. Because of insufficient cytoreductive efficacy, the dose was increased to 1,500 mg five days later. Eight days after the initiation of hydroxyurea, the patient started complaining of chills, fever, and vomiting. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were markedly elevated to 5,098 and 3,880 IU/l from 44 and 59 IU/l in one day, respectively. Tests for hepatitis viruses were all negative. With the discontinuation of hydroxyurea, AST and ALT returned to their former levels within two weeks. A drug-induced lymphocyte stimulation test for hydroxyurea was positive with a stimulating index of 2.0. Hepatic dysfunction has been recognized as one of the side effects of hydroxyurea. However, there have been only a limited number of reports demonstrating drug allergy to have a role in hepatic dysfunction accompanied by fever and gastrointestinal symptoms. The findings of our case strongly suggest that all presentations could be explained by drug allergy. Physicians should be mindful of the potential for acute and severe hepatic dysfunction due to allergic reaction against hydroxyurea.


An analytical bond-order potential for carbon.
Carbon is the most widely studied material today because it exhibits special properties not seen in any other materials when in nano dimensions such as nanotube and graphene. Reduction of material defects created during synthesis has become critical to realize the full potential of carbon structures. Molecular dynamics (MD) simulations, in principle, allow defect formation mechanisms to be studied with high fidelity, and can, therefore, help guide experiments for defect reduction. Such MD simulations must satisfy a set of stringent requirements. First, they must employ an interatomic potential formalism that is transferable to a variety of carbon structures. Second, the potential needs to be appropriately parameterized to capture the property trends of important carbon structures, in particular, diamond, graphite, graphene, and nanotubes. Most importantly, the potential must predict the crystalline growth of the correct phases during direct MD simulations of synthesis to achieve a predictive simulation of defect formation. Because an unlimited number of structures not included in the potential parameterization are encountered, the literature carbon potentials are often not sufficient for growth simulations. We have developed an analytical bond order potential for carbon, and have made it available through the public MD simulation package LAMMPS. We demonstrate that our potential reasonably captures the property trends of important carbon phases. Stringent MD simulations convincingly show that our potential accounts not only for the crystalline growth of graphene, graphite, and carbon nanotubes but also for the transformation of graphite to diamond at high pressure.


Female Narcissism: Assessment, Aetiology, and Behavioural Manifestations.
Despite putative gender differences in the expression of narcissism, prominent theories have virtually dismissed the role of females in the development and manifestation of narcissism. The contention that narcissism is a pathology of the self that may partly differ in males and females is further evident in the Diagnostic and Statistical Manual of Mental Disorders (DSM-5). The DSM-5 reports that up to 75% of those diagnosed with Narcissistic Personality Disorder (NPD) are men. Such figures suggest that the representation of narcissism as codified in the DSM-5 may only be marginally applicable to females, given its prominent focus and nature on capturing grandiose themes which closely resemble commonly masculine norms. The overemphasis on grandiose features extends to the empirical literature which defines narcissism as a normative personality trait and is widely assessed using the Narcissistic Personality Inventory (NPI), on which males obtain significantly higher scores than females. As this review will demonstrate, one limitation frequently occurring in the literature is the attempt to comprehend narcissistic manifestations in females through the lens of what has commonly been defined as narcissism (DSM/NPI). In this review, the literature concerning the diagnostic assessment and conceptualisation of narcissistic personality disorder, aetiological factors, aggression, and partner violence perpetration will be discussed in relation to the importance of gender. This is followed by a review of existing gaps in theory and research, and suggestions for fruitful directions that can aid a richer and more meaningful literature on narcissism inclusive of gender issues.


Performance of idarucizumab as antidote of dabigatran in daily clinical practice.
Because practice-based data on the usage of idarucizumab for urgent dabigatran reversal is unavailable, we evaluated the appropriateness of idarucizumab usage, its haemostatic effectiveness and clinical outcomes. An observational cohort study was performed including consecutive patients who were treated with idarucizumab between 2016 and 2018. Appropriate usage was assessed with predefined criteria. Post-reversal effectiveness was evaluated according to International Society on Thrombosis and Haemostasis (ISTH) recommendations. Patients were followed for 90 days for occurrence of thromboembolism, (re-)bleeding and death. Idarucizumab was used in 88 patients, of whom 53 (60%) presented with severe bleeding (20 gastrointestinal and 18 intracranial) and 35 (40%) requiring urgent surgical intervention. Use of idarucizumab was judged inappropriate in 25 patients (28%). Effective haemostasis was achieved in 32 of 48 (67%) bleeding patients in whom assessment was possible. Seven of 16 patients with major bleeding who did not achieve effective haemostasis (five intracranial) died, compared with two of 32 patients with effective haemostasis (relative risk 7.0, 95% confidence interval 1.6-30). Four patients (4.2%) developed thromboembolism [2 (2.1%) within 30 days] and four patients (4.2%) re-bleeding, all within 10 days. Seventeen patients (19%) died; 10 (11%) within 5 days. In this practice-based cohort, idarucizumab use was considered inappropriate in 28% of patients. Effective haemostasis was achieved in two-third of bleeding patients and was associated with lower mortality risk. Clinical outcomes were similar to those observed in the RE-VERSE AD trial, comprising re-bleeds and thromboembolism, and a high-mortality rate.


Anterior forebrain pathway in parrots is necessary for producing learned vocalizations with individual signatures.
Parrots have enormous vocal imitation capacities and produce individually unique vocal signatures. Like songbirds, parrots have a nucleated neural song system with distinct anterior (AFP) and posterior forebrain pathways (PFP). To test if song systems of parrots and songbirds, which diverged over 50 million years ago, have a similar functional organization, we first established a neuroscience-compatible call-and-response behavioral paradigm to elicit learned contact calls in budgerigars (Melopsittacus undulatus). Using variational autoencoder-based machine learning methods, we show that contact calls within affiliated groups converge but that individuals maintain unique acoustic features, or vocal signatures, even after call convergence. Next, we transiently inactivated the outputs of AFP to test if learned vocalizations can be produced by the PFP alone. As in songbirds, AFP inactivation had an immediate effect on vocalizations, consistent with a premotor role. But in contrast to songbirds, where the isolated PFP is sufficient to produce stereotyped and acoustically normal vocalizations, isolation of the budgerigar PFP caused a degradation of call acoustic structure, stereotypy, and individual uniqueness. Thus, the contribution of AFP and the capacity of isolated PFP to produce learned vocalizations have diverged substantially between songbirds and parrots, likely driven by their distinct behavioral ecology and neural connectivity.


Efficacy of Electrical Stimulators for Bone Healing: A Meta-Analysis of Randomized Sham-Controlled Trials.
Electrical stimulation is a common adjunct used to promote bone healing; its efficacy, however, remains uncertain. We conducted a meta-analysis of randomized sham-controlled trials to establish the efficacy of electrical stimulation for bone healing. We identified all trials randomizing patients to electrical or sham stimulation for bone healing. Outcomes were pain relief, functional improvement, and radiographic nonunion. Two reviewers assessed eligibility and risk of bias, performed data extraction, and rated the quality of the evidence. Fifteen trials met our inclusion criteria. Moderate quality evidence from 4 trials found that stimulation produced a significant improvement in pain (mean difference (MD) on 100-millimeter visual analogue scale = -7.7 mm; 95% CI -13.92 to -1.43; p = 0.02). Two trials found no difference in functional outcome (MD = -0.88; 95% CI -6.63 to 4.87; p = 0.76). Moderate quality evidence from 15 trials found that stimulation reduced radiographic nonunion rates by 35% (95% CI 19% to 47%; number needed to treat = 7; p < 0.01). Patients treated with electrical stimulation as an adjunct for bone healing have less pain and are at reduced risk for radiographic nonunion; functional outcome data are limited and requires increased focus in future trials.


Factor XIIIA Val34Leu polymorphism does not predict risk of coronary heart disease: The Atherosclerosis Risk in Communities (ARIC) Study.
Factor XIII catalyzes the cross-linking of fibrin. Cross-sectional studies have suggested that a common polymorphism site at residue 34 of the A subunit of factor XIII (FXIIIA) with a substitution of Leu for Val (FXIIIA Val34Leu) was associated with reduced coronary heart disease (CHD). This association has not been examined in prospective studies. Healthy subjects (n=15 792) were recruited from 4 US communities into the Atherosclerosis Risk in Communities (ARIC) study from 1987 to 1989. They were followed, and incident CHD events were identified and verified. For the present study, we included 423 patients with CHD as cases and 479 noncases. FXIIIA Val34Leu polymorphism was determined by the single-strand conformational polymorphism method. There were no significant differences in the genotype frequencies between cases and noncases. The genotypes showed little association with known CHD risk factors. A weighted proportional hazards regression analysis adjusting for other risk factors confirmed no association of the genotypes with risk of CHD. This prospective study did not provide evidence of a reduced CHD risk for the FXIIIA 34Leu allele.


Proteasome Subunits Differentially Control Myeloma Cell Viability and Proteasome Inhibitor Sensitivity.
We generated eight multiple myeloma cell lines resistant to bortezomib; five acquired PSMB5 mutations. In 1,500 patients such mutations were rare clinically. To better understand disruption of proteasomes on multiple myeloma viability and drug sensitivity, we systematically deleted the major proteasome catalytic subunits. Multiple myeloma cells without PSMB5 were viable. Drug-resistant, PSMB5-mutated cell lines were resensitized to bortezomib by PSMB5 deletion, implying PSMB5 mutation is activating in its drug resistance function. In contrast, PSMB6 knockout was lethal to multiple myeloma cell lines. Depleting PSMB6 prevented splicing of the major catalytic subunits PSMB5, PSMB7, PSMB8, and PSMB10; however, PSMB6 engineered without splicing function or catalytic activity, also restored viability, inferring the contribution of PSMB6 to proteasome structure to be more important than functional activity. Supporting this, bortezomib sensitivity was restored in drug-resistant multiple myeloma cell lines by low level expression of mutated PSMB6 lacking splicing function. Loss of PSMB8 and PSMB9 was neither lethal nor restored bortezomib sensitivity. Significant codependency of PSMB5, PSMB6, and PSMB7 expression was observed. We demonstrated elevated levels of PSMB6 and 7, but not 8 and 9, in some, but not all, serial patient samples exposed to proteasome inhibitors. In summary, we show PSMB6 and PSMB7, but not PSMB5, to be essential for multiple myeloma cell survival, this dependency is structural and that upregulation or activating mutation of PSMB5, 6, and 7 confers proteasome inhibitor resistance, while depletion confers sensitivity. IMPLICATIONS: These findings support modulation of PSMB5, PSMB6, or PSMB7 expression as a new therapeutic strategy.


Aberrant O-glycosylation inhibits stable expression of dysadherin, a carcinoma-associated antigen, and facilitates cell-cell adhesion.
Recently, we identified dysadherin, a novel carcinoma-associated glycoprotein, and showed that overexpression of dysadherin in human hepatocarcinoma PLC/PRF/5 cells could suppress E-cadherin-mediated cell-cell adhesion and promote tumor metastasis. The present study shows evidence that dysadherin is actually O-glycosylated. This was based on a direct carbohydrate composition analysis of a chimera protein of an extracellular domain of dysadherin fused to an Fc fragment of immunoglobulin. To assess the importance of O-glycosylation in dysadherin function, dysadherin-transfected hepatocarcinoma cells were cultured in a medium containing benzyl-alpha-GalNAc, a modulator of O-glycosylation. This treatment facilitated homotypic cell adhesion among dysadherin transfectants accompanied with morphological changes, indicating that the anti-adhesive effect of dysadherin was weakened. Modification of O-glycan synthesis also resulted in down-regulation of dysadherin expression and up-regulation of E-cadherin expression in dysadherin transfectants but did not affect E-cadherin expression in mock transfectants. Structural analysis of O-glycans released from the dysadherin chimera proteins indicated that a series of O-glycans with core 1 and 2 structures are attached to dysadherin, and their sialylation is remarkably inhibited by benzyl-alpha-GalNAc treatment. However, sialidase treatment of the cells did not affect calcium-dependent cell aggregation, which excluded the possibility that sialic acid itself is directly involved in cell-cell adhesion. We suggest that aberrant O-glycosylation in carcinoma cells inhibits stable expression of dysadherin and leads to the up-regulation of E-cadherin expression by an unknown mechanism, resulting in increased cell-cell adhesion. The carbohydrate-directed approach to the regulation of dysadherin expression might be a new strategy for cancer therapy.