Inactivation of tautomerase activity of macrophage migration inhibitory factor by sulforaphane: a potential biomarker for anti-inflammatory intervention.
Macrophage migration inhibitory factor (MIF), a proinflammatory cytokine with keto-enol tautomerase activity, rises rapidly in response to inflammation and is elevated in many chronic diseases. Isothiocyanates, such as sulforaphane from broccoli, are very potent inactivators of MIF tautomerase activity. A simple rapid method for determining this activity in tissues and body fluids may therefore be valuable for assessing severity of inflammation and efficacy of intervention. Existing spectrophotometric assays of MIF, based on conversion of methyl L-dopachrome to methyl 5,6-dihydroxyindole-2-carboxylate and associated loss of absorption at 475 nm, lack sensitivity. Assay sensitivity and efficiency were markedly improved by reducing the nonenzymatic rate, by lowering pH to 6.2, replacing phosphate (which catalyzes the reaction) with Bis-Tris buffer, and converting to a microtiter plate format. A structure-potency study of MIF tautomerase inactivation by isothiocyanates showed that sulforaphane, benzyl, n-hexyl, and phenethyl isothiocyanates were especially potent. MIF tautomerase could be readily quantified in human urine concentrated by ultrafiltration. This activity comprised: (i) a heat-labile, sulforaphane-inactivated macromolecular fraction (presumably MIF) that was concentrated during ultrafiltration; (ii) a flow-through fraction, with constant activity during filtration, that was heat stable and insensitive to sulforaphane. Administration of the sulforaphane precursor glucoraphanin to human volunteers almost completely abolished urinary tautomerase activity, which recovered over many hours. A simple, rapid, quantitative MIF tautomerase assay has been developed as a potential biomarker for assessing inflammatory severity and effectiveness of intervention. An improved assay for measuring MIF tautomerase activity and its applications are described.


Strategic origins of early semantic facilitation in the blocked-cyclic naming paradigm.
In the blocked-cyclic naming paradigm, participants repeatedly name small sets of objects that do or do not belong to the same semantic category. A standard finding is that, after a first presentation cycle where one might find semantic facilitation, naming is slower in related (homogeneous) than in unrelated (heterogeneous) sets. According to competitive theories of lexical selection, this is because the lexical representations of the object names compete more vigorously in homogeneous than in heterogeneous sets. However, Navarrete, del Prato, Peressotti, and Mahon (2014) argued that this pattern of results was not due to increased lexical competition but to weaker repetition priming in homogeneous compared to heterogeneous sets. They demonstrated that when homogeneous sets were not repeated immediately but interleaved with unrelated sets, semantic relatedness induced facilitation rather than interference. We replicate this finding but also show that the facilitation effect has a strategic origin: It is substantial when sets are separated by pauses, making it easy for participants to notice the relatedness within some sets and use it to predict upcoming items. However, the effect is much reduced when these pauses are eliminated. In our view, the semantic facilitation effect does not constitute evidence against competitive theories of lexical selection. It can be accounted for within any framework that acknowledges strategic influences on the speed of object naming in the blocked-cyclic naming paradigm. (PsycINFO Database Record


Precision methylome characterization of Mycobacterium tuberculosis complex (MTBC) using PacBio single-molecule real-time (SMRT) technology.
Tuberculosis (TB) remains one of the most common infectious diseases caused by Mycobacterium tuberculosis complex (MTBC). To panoramically analyze MTBC's genomic methylation, we completed the genomes of 12 MTBC strains (Mycobacterium bovis; M. bovis BCG; M. microti; M. africanum; M. tuberculosis H37Rv; H37Ra; and 6 M. tuberculosis clinical isolates) belonging to different lineages and characterized their methylomes using single-molecule real-time (SMRT) technology. We identified three (m6)A sequence motifs and their corresponding methyltransferase (MTase) genes, including the reported mamA, hsdM and a newly discovered mamB. We also experimentally verified the methylated motifs and functions of HsdM and MamB. Our analysis indicated the MTase activities varied between 12 strains due to mutations/deletions. Furthermore, through measuring 'the methylated-motif-site ratio' and 'the methylated-read ratio', we explored the methylation status of each modified site and sequence-read to obtain the 'precision methylome' of the MTBC strains, which enabled intricate analysis of MTase activity at whole-genome scale. Most unmodified sites overlapped with transcription-factor binding-regions, which might protect these sites from methylation. Overall, our findings show enormous potential for the SMRT platform to investigate the precise character of methylome, and significantly enhance our understanding of the function of DNA MTase.


Using graphics processors to accelerate protein docking calculations.
Protein docking is the computationally intensive task of calculating the three-dimensional structure of a protein complex starting from the individual structures of the constituent proteins. In order to make the calculation tractable, most docking algorithms begin by assuming that the structures to be docked are rigid. This article describes some recent developments we have made to adapt our FFT-based "Hex" rigid-body docking algorithm to exploit the computational power of modern graphics processors (GPUs). The Hex algorithm is very efficient on conventional central processor units (CPUs), yet significant further speed-ups have been obtained by using GPUs. Thus, FFT-based docking calculations which formerly took many hours to complete using CPUs may now be carried out in a matter of seconds using GPUs. The Hex docking program and access to a server version of Hex on a GPU-based compute cluster are both available for public use.


Comprehensive Assessment of Fine-Grained Wound Images Using a Patch-Based CNN With Context-Preserving Attention.
Chronic wounds affect 6.5 million Americans. Wound assessment via algorithmic analysis of smartphone images has emerged as a viable option for remote assessment. We comprehensively score wounds based on the clinically-validated Photographic Wound Assessment Tool (PWAT), which comprehensively assesses clinically important ranges of eight wound attributes: Size, Depth, Necrotic Tissue Type, Necrotic Tissue Amount, Granulation Tissue type, Granulation Tissue Amount, Edges, Periulcer Skin Viability. We proposed a DenseNet Convolutional Neural Network (CNN) framework with patch-based context-preserving attention to assess the 8 PWAT attributes of four wound types: diabetic ulcers, pressure ulcers, vascular ulcers and surgical wounds. In an evaluation on our dataset of 1639 wound images, our model estimated all 8 PWAT sub-scores with classification accuracies and F1 scores of over 80%. Our work is the first intelligent system that autonomously grades wounds comprehensively based on criteria in the PWAT rubric, alleviating the significant burden that manual wound grading imposes on wound care nurses.


Seasonal variations in the harpagoside content of Scrophularia scorodonia L.
Seasonal variations on the content of harpagoside in Scrophularia scorodonia L. (Scrophulariaceae) were investigated using plants collected monthly from January to December in 1995. During growth of this species the percentage of harpagoside was the highest during the maximum development of the plant, specially in July. Harpagoside levels differed among leaves, stems and flowers of S. scorodonia. Leaves were distinguished from other plant parts by higher levels of harpagoside. Drying at ambient temperature influenced the yield of harpagoside compared with the results of plant drying by microwave.


A Label-Free Platform for Identification of Exosomes from Different Sources.
Exosomes contain cell- and cell-state-specific cargos of proteins, lipids, and nucleic acids and play significant roles in cell signaling and cell-cell communication. Current research into exosome-based biomarkers has relied largely on analyzing candidate biomarkers, i.e., specific proteins or nucleic acids. However, this approach may miss important biomarkers that are yet to be identified. Alternative approaches are to analyze the entire exosome system, either by "omics" methods or by techniques that provide "fingerprints" of the system without identifying each individual biomolecule component. Here, we describe a platform of the latter type, which is based on surface-enhanced Raman spectroscopy (SERS) in combination with multivariate analysis, and demonstrate the utility of this platform for analyzing exosomes derived from different biological sources. First, we examined whether this analysis could use exosomes isolated from fetal bovine serum using a simple, commercially available isolation kit or necessitates the higher purity achieved by the "gold standard" ultracentrifugation/filtration procedure. Our data demonstrate that the latter method is required for this type of analysis. Having established this requirement, we rigorously analyzed the Raman spectral signature of individual exosomes using a unique, hybrid SERS substrate made of a graphene-covered Au surface containing a quasi-periodic array of pyramids. To examine the source of the Raman signal, we used Raman mapping of low and high spatial resolution combined with morphological identification of exosomes by scanning electron microscopy. Both approaches suggested that the spectra were collected from single exosomes. Finally, we demonstrate for the first time that our platform can distinguish among exosomes from different biological sources based on their Raman signature, a promising approach for developing exosome-based fingerprinting. Our study serves as a solid technological foundation for future exploration of the roles of exosomes in various biological processes and their use as biomarkers for disease diagnosis and treatment monitoring.


Effects of long-term exposure to reduced pH conditions on the shell and survival of an intertidal gastropod.
Volcanic CO2 vents are useful environments for investigating the biological responses of marine organisms to changing ocean conditions (Ocean acidification, OA). Marine shelled molluscs are highly sensitive to changes in seawater carbonate chemistry. In this study, we investigated the effects of reduced pH on the intertidal gastropod, Phorcus sauciatus, in a volcanic CO2 vent off La Palma Island (Canary Islands, North East Atlantic Ocean), a location with a natural pH gradient ranging from 7.0 to 8.2 over the tidal cycles. Density and size-frequency distribution, shell morphology, shell integrity, fracture resistance, and desiccation tolerance were evaluated between populations from control and CO2 vent sites. We found no effects of reduced pH on population parameters or desiccation tolerance across the pH gradient, but significant differences in shell morphology, shell integrity, and fracture resistance were detected. Individuals from the CO2 vent site exhibited a higher shell aspect ratio, greater percentages of shell dissolution and break, and compromised shell strength than those from the control site. Our results highlight that long-term exposure to high pCO2 can negatively affect the shell features of P. sauciatus but may not have a significant effect on population performance. Moreover, we suggest that loss of shell properties could lead to changes in predator-prey interactions.


Oromandibular Dystonia Related to Sertraline Treatment in a Child.
In children, similar to adults, the agents that cause movement disorders are most commonly dopaminergic and antidopaminergic drugs. However, it is also known that selective serotonin reuptake inhibitors (SSRIs), which are frequently prescribed in child psychiatry practice, may rarely lead to tremor, akathisia, dystonia, dyskinesia, and parkinsonism. In this study, we report a rare side effect of SSRIs, namely oromandibular dystonia related to sertraline, in a pediatric patient who was diagnosed with posttraumatic stress disorder.


Association of Mediterranean diet and cardiorespiratory fitness with the development of pre-diabetes and diabetes: the Coronary Artery Risk Development in Young Adults (CARDIA) study.
To better understand the association between a modified Mediterranean diet pattern in young adulthood, cardiorespiratory fitness in young adulthood, and the odds of developing pre-diabetes or diabetes by middle age. Participants from the Coronary Artery Risk Development in Young Adults (CARDIA) study who did not have pre-diabetes or diabetes at baseline (year 0 (Y0), ages 18-30) and who had data available at the Y0 and year 25 (Y25) visits were included in this analysis (n=3358). Polytomous logistic regression models were used to assess the association between baseline dietary intake and fitness data and odds of pre-diabetes or diabetes by middle age (Y25, ages 43-55). At the Y25 visit, 1319 participants (39%) had pre-diabetes and 393 (12%) had diabetes. Higher baseline fitness was associated with lower odds of pre-diabetes and of diabetes at Y25. After adjustment for covariates, each SD increment in treadmill duration (181 s) was associated with lower odds for pre-diabetes (OR 0.85, 95% CI 0.75 to 0.95, p=0.005) and for diabetes (OR 0.71, 95% CI 0.60 to 0.85, p=0.0002) when compared to normal glycemia. A modified Mediterranean diet pattern was not associated with either pre-diabetes or diabetes. No interaction between cardiorespiratory fitness and dietary intake was observed, but baseline fitness remained independently associated with incident pre-diabetes and diabetes following adjustment for diet. Higher cardiorespiratory fitness in young adulthood, but not a modified Mediterranean diet pattern, is associated with lower odds of pre-diabetes and of diabetes in middle age. NCT00005130.


Effects of acetylsalicylic acid on sore throat pain and other pain symptoms associated with acute upper respiratory tract infection.
Acetylsalicylic acid (ASA) has been widely used for over a century to treat pain and fever associated with acute upper respiratory tract infection (URTI), but there is a lack of clinical data to support the efficacy of ASA in this disease state. The objective of this study was to investigate the efficacy and safety of ASA for the treatment of sore throat pain associated with URTI. A double-blinded, placebo-controlled, parallel group design. Two hundred seventy-two patients (mean age: 25 years) with sore throat pain associated with URTI were recruited at two centers. Pain scores were made during a 2-hour laboratory phase and continued for secondary objectives during a 4-hour home phase. Patients were treated with either two effervescent tablets of ASA 400 mg in water or matched placebo tablets. Patients took medication as required over a 3-day home phase. ASA was found to be superior to placebo for: The primary efficacy parameter predefined in the protocol, reduction in sore throat pain intensity over 2 hours (P < 0.001), and for secondary efficacy parameters, reduction in sore throat pain intensity over 4 and 6 hours, relief of sore throat pain over 2, 4, and 6 hours, reduction in intensity of pain associated with headache, and reduction in muscle aches and pains over a 2-hour time period (P < 0.01). No safety problems were encountered. Treatment with ASA was shown to provide relief from sore throat pain, headache, and muscle aches and pains associated with URTI.


Sex-specific risk profiles for suicide mortality in bipolar disorder: incidence, healthcare utilization and comorbidity.
Evidence on sex-specific incidence and comorbidity risk factors of suicide among patients with bipolar disorder is scarce. This study investigated the sex-specific risk profiles for suicide among the bipolar disorder population in terms of incidence, healthcare utilization and comorbidity. Using data from the Taiwan National Health Insurance Research Database between 1 January 2000 and 31 December 2016, this nationwide cohort study included patients with bipolar disorder (N = 46 490) and individuals representative of the general population (N = 185 960) matched by age and sex at a 1:4 ratio. Mortality rate ratios (MRRs) of suicide were calculated between suicide rates of bipolar disorder cohort and general population. In addition, a nested case-control study (1428 cases died by suicide and 5710 living controls) was conducted in the bipolar disorder cohort to examine the sex-specific risk of healthcare utilization and comorbidities. Suicide risk was considerably higher in the cohort (MRR = 21.9) than in the general population, especially among women (MRR = 35.6). Sex-stratified analyses revealed distinct healthcare utilization patterns and physical comorbidity risk profiles between the sexes. Although female patients who died by suicide had higher risks of nonhypertensive cardiovascular disease, pneumonia, chronic kidney disease, peptic ulcer, irritable bowel syndrome, and sepsis compared to their living counterparts, male patients who died by suicide had higher risks of chronic kidney disease and sepsis compared to the living controls. Patients with bipolar disorder who died by suicide had sex-specific risk profiles in incidence and physical comorbidities. Identifying these modifiable risk factors may guide interventions for suicide risk reduction.


Thermodynamic and structural equivalence of two HLA-B27 subtypes complexed with a self-peptide.
The F pocket of major histocompatibility complex (in humans HLA) class I molecules accommodates the C terminus of the bound peptide. Residues forming this pocket exhibit considerable polymorphism, and a single difference (Asp116 in HLA-B*2705 and His116 in HLA-B*2709 heavy chains) confers differential association of these two HLA-B27 subtypes to the autoimmune disease ankylosing spondylitis. As peptide presentation by HLA molecules is of central importance for immune responses, we performed thermodynamic (circular dichroism, differential scanning calorimetry, fluorescence polarization) and X-ray crystallographic analyses of both HLA-B27 subtypes complexed with the epidermal growth factor response factor 1-derived self-peptide TIS (RRLPIFSRL) to understand the impact of the Asp116His exchange on peptide display. This peptide is known to be presented in vivo by both subtypes, and as expected for a self-peptide, TIS-reactive cytotoxic T lymphocytes are absent in the respective individuals. The thermodynamic analyses reveal that both HLA-B27:TIS complexes exhibit comparable, relatively high thermostability (Tm approximately 60 degrees C) and undergo multi-step unfolding reactions, with dissociation of the peptide in the first step. As shown by X-ray crystallography, only subtle structural differences between the subtypes were observed regarding the architecture of their F pockets, including the presence of distinct networks of water molecules. However, no consistent structural differences were found between the peptide presentation modes. In contrast to other peptides displayed by the two HLA-subtypes which show either structural or dynamical differences in their peptide presentation modes, the TIS-complexed HLA-B*2705 and HLA-B*2709 subtypes are an example for thermodynamic and structural equivalence, in agreement with functional data.


Stability patterns for a size-structured population model and its stage-structured counterpart.
In this paper we compare a general size-structured population model, where a size-structured consumer feeds upon an unstructured resource, to its simplified stage-structured counterpart in terms of equilibrium stability. Stability of the size-structured model is understood in terms of an equivalent delayed system consisting of a renewal equation for the consumer population birth rate and a delayed differential equation for the resource. Results show that the size- and stage-structured models differ considerably with respect to equilibrium stability, although the two models have completely identical equilibrium solutions. First, when adult consumers are superior foragers to juveniles, the size-structured model is more stable than the stage-structured model while the opposite occurs when juveniles are the superior foragers. Second, relatively large juvenile (adult) mortality tends to stabilise (destabilise) the size-structured model but destabilise (stabilise) the stage-structured model. Third, the stability pattern is sensitive to the adult-offspring size ratio in the size-structured model but much less sensitive in the stage-structured model. Finally, unless the adult-offspring size ratio is sufficiently small, the stage-structured model cannot satisfactorily capture the dynamics of the size-structured model. We conclude that caution must be taken when the stage-structured population model is applied, although it can consistently translate individual life history and stage-specific differences to the population level.


SOCIOEMOTIONAL AND BEHAVIORAL PROBLEMS IN TODDLERS WITH LANGUAGE DELAY.
Toddlers with language delay are at risk for persistent developmental and behavioral difficulties; however, the association between socioemotional/behavior problems and language in young children is not well understood. This study explored socioemotional/behavior problems in a unique sample of toddlers with language delays using a measure developed explicitly for this age group. Toddlers identified by 18 months with receptive and expressive language delay (LD; n = 30) or typical development (TD; n = 61) were evaluated at 18 and 24 months of age using the Infant-Toddler Social and Emotional Assessment (ITSEA) and the Mullen Scales of Early Learning. Compared to toddlers who had TD, toddlers with LD had significantly more concerning scores at 18 and 24 months on all ITSEA domains. The rate of "clinical concern" on most domains was not high in either group, except that >60% of LD toddlers were in the clinical concern range on the Competence domain. Socioemotional/behavioral problems were dimensionally related to receptive and expressive language, with greater language delay associated with more concerning ITSEA scores. Socioemotional and behavioral problems are related to receptive and expressive language abilities in 18- and 24-month-olds, indicating the need for screening of both types of concerns in toddlers identified with potential language delays.


[Assessing social inequality in microcensus data and German national health examination survey].
To analyse the impact of social inequality on health and illness public health officials in Germany request for more data on socio-demographic indicators in official population statistics at Federal and State level. The aim of the study was to examine whether the national sample census Microcensus is suitable for scaling social inequality. For this purpose, coding instructions for calculating social strata index by Winkler were applied to databases of the Microcensus. The results were compared with the German Health Survey, because social data of this survey are already scaled according to the coding instruction for social strata index by Winkler. Data analyses were based on subgroups of North Rhine-Westphalia originated from national samples of Microcensus and German Health Survey. For index indicators "education" and "income" coding structure was transferred to databases of Microcensus without any problems. For index indicator "occupational status" occupational groups were less comparable. This refers to the classification of the occupational status of formerly employed people. However this limitation applies also to employees because the model for "occupational status" is assessed only on a four-year basis. From a methodical point of view it seems to be a problem that "household" has a different meaning in the index by Winkler and Microcensus and is only partly transferable to Microcensus databases. Additionally, identification of the main income earner of the household proved extremely difficult. Therefore, at the present conceptional stage of Microcensus it seems appropriate to classify occupational status as an individual criterion.


Are we asking the right questions? A review of Canadian REB practices in relation to community-based participatory research.
Access barriers to effective ethics review continue to be a significant challenge for researchers and community-based organizations undertaking community-based participatory research (CBPR). This article reports on findings from a content analysis of select (Behavioural, Biomedical, Social Sciences, Humanities) research ethics boards (REBs) in the Canadian research context (n = 86). Existing ethics review documentation was evaluated using 30 CBPR related criteria for their sensitivity to relevant approaches, processes, and outcomes. A linear regression was conducted to determine whether specific organizational characteristics have an impact on the CBPR sensitivity: (1) region of Canada, (2) type of institution (university or a healthcare organization), (3) primary institutional language (English or French) and (4) national ranking with respect to research intensiveness. While only research intensiveness proved statistically significant (p = .001), we recognize REB protocol forms may not actually reflect how CBPR is reviewed. Despite using a single guiding ethical framework, REBs across Canada employ a variety of techniques to review research studies. We report on these differences and varying levels of sensitivity to CBPR. Finally, we highlight best practices and make recommendations for integrating CBPR principles into existing ethics review.


European Good Laboratory and Clinical Practices: their relevance to clinical pathology laboratories.
The requirements for Good Laboratory (GLP) and Good Clinical Practices (CGP) were established as a matter of urgency by the United States in the early 1970s. These were in response to gross misconduct and, in many instances, fraud. Over the next 15 years, a plethora of regulatory principles, guidelines, and regulations was produced by many countries of the world, culminating in single standards for European, Japanese, and United States authorities. Although with regard to GLP this has basically become a worldwide recognized standard within the preclinical (toxicology) studies, in the veterinary, chemical, agrochemical, and pharmaceutical industries, the GCPs are now seeing a rebirth. Within a clinical trials environment, there is most certainly a requirement for compliance with GCP, especially with regard to the harmonization of data within the European Community. The goal of this article is to cover the following aspects: Why should we have good practices? Why should laboratory data be audited? Why is there a need for a QA unit or function? What is the QA operational approach? How does a laboratory audit take place within laboratories? In discussing the laboratories and their subsequent data audits, the pitfalls and benefits are addressed and an examination of the data from the sponsor's viewpoint is compared with that produced by the laboratory. The types of laboratories present in a clinical environment are examined. They obviously comprise clinical pathology, microbiology, and analytical as well as ancillary hospital areas such as X-ray and cardiology. These laboratories may also be in the private sector, the National Health Service, contract laboratories, universities, or the general practitioner population.(ABSTRACT TRUNCATED AT 250 WORDS)


Transfusion dependence, use of hospice services, and quality of end-of-life care in leukemia.
Hospice provides high-quality end-of-life care, but patients with leukemias use hospice services less frequently than those with solid tumors. Transfusion dependence (TD) may hinder or delay enrollment, because hospice organizations typically disallow transfusions. We examined the association between TD and end-of-life outcomes among Medicare beneficiaries with leukemia. From the Surveillance, Epidemiology, and End Results-Medicare database, we selected beneficiaries with acute and chronic leukemias who died in 2001-2011. We defined TD as ≥2 transfusions within 30 days before death or hospice enrollment. End points included hospice enrollment and length of stay, reporting relative risk (RR) adjusted for key covariates. Among 21 033 patients with a median age of 79 years, 20% were transfusion dependent before death/hospice enrollment. Use of hospice increased from 35% in 2001 to 49% in 2011. Median time on hospice was 9 days and was shorter for transfusion-dependent patients (6 vs 11 days; P < .001). Adjusting for baseline characteristics, TD was associated with a higher use of hospice services (RR, 1.08; 95% confidence interval [CI], 1.04-1.12) but also with 51% shorter hospice length of stay (RR, 0.49; 95% CI, 0.44-0.54). Hospice enrollees had a lower likelihood of inpatient death and chemotherapy use and lower median Medicare spending at end-of-life, regardless of TD status. In conclusion, relatively increased hospice use combined with a markedly shorter length of stay among transfusion-dependent patients suggests that they have a high and incompletely met need for hospice services and that they experience a barrier to timely referral. Policy solutions supporting palliative transfusions may maximize the benefits of hospice for leukemia patients.


Influence of mucinous components on survival in colorectal adenocarcinomas: a multivariate analysis.
Of 534 resected colorectal adenocarcinomas, 165 (31%) contained some mucinous components; these represented the main part of the tumour in 67 (13%). Of the mucin containing tumours, 63 (38%) were in the right colon compared with 50 (13%) of the non-mucinous ones (p less than 0.001). Patients with predominantly mucinous tumours were significantly older than those with non-mucinous tumours, and they tended to present with tumours at a more advanced stage. A multivariate analysis did not show any significant independent prognostic influence of the mucinous component except when this had a predominantly signet ring cell pattern.


A p38MAPK-p53 cascade regulates mesodermal differentiation and neurogenesis of embryonic stem cells.
Embryonic stem cells (ESCs) differentiate in vivo and in vitro into all cell lineages, and they have been proposed as cellular therapy for human diseases. However, the molecular mechanisms controlling ESC commitment toward specific lineages need to be specified. We previously found that the p38 mitogen-activated protein kinase (p38MAPK) pathway inhibits neurogenesis and is necessary to mesodermal formation during the critical first 5 days of mouse ESC commitment. This period corresponds to the expression of specific master genes that direct ESC into each of the three embryonic layers. By both chemical and genetic approaches, we found now that, during this phase, the p38MAPK pathway stabilizes the p53 protein level and that interfering directly with p53 mimics the effects of p38MAPK inhibition on ESC differentiation. Anti-p53 siRNA transient transfections stimulate Bcl2 and Pax6 gene expressions, leading to increased ESC neurogenesis compared with control transfections. Conversely, p53 downregulation leads to a strong inhibition of the mesodermal master genes Brachyury and Mesp1 affecting cardiomyogenesis and skeletal myogenesis of ESCs. Similar results were found with p53(-/-) ESCs compared with their wild-type counterparts. In addition, knockout p53 ESCs show impaired smooth muscle cell and adipocyte formation. Use of anti-Nanog siRNAs demonstrates that certain of these regulations result partially to p53-dependent repression of Nanog gene expression. In addition to its well-known role in DNA-damage response, apoptosis, cell cycle control and tumor suppression, p53 has also been involved in vivo in embryonic development; our results show now that p53 mediates, at least for a large part, the p38MAPK control of the early commitment of ESCs toward mesodermal and neural lineages.


A reliability study of dynamic and static pain tests in temporomandibular disorder patients.
To determine the interexaminer reliability of dynamic and static pain tests in patients with temporomandibular disorders (TMD). One hundred fifteen consecutive TMD patients participated in the study. At intake, pain on dynamic and static pain tests was scored on a 4-point ordinal scale by 1 of 5 dentists. Pressure was applied to the mandible during mandibular opening, closing, and protrusive movements (dynamic tests) and while the mandible was kept motionless by the patient in an open, closed, or protrusive position (static tests). After this examination, the dynamic and static pain tests were performed a second time by 1 of 2 physical therapists blinded to the outcome of the first examination. Prior to the study, all examiners took part in a yearly training session, while 3 examiners (2 dentists and 1 physical therapist) were trained on a more regular basis. The interexaminer reliability of dynamic and static pain tests ranged from "poor" to "fair to good" (intraclass correlation coefficient [ICC]: 0.29-0.54) but reached the "excellent" level (ICC: 0.34-0.92) when only the data gathered by the more extensively trained examiners were considered. The reliability was higher when the data were analyzed on the 4-point scale as compared to a dichotomized pain scale. The reliability of dynamic and static pain tests for the temporomandibular region is fair to good when rated on an ordinal pain scale. Thorough training of examiners can improve the reliability considerably.


GC-MS based metabolomics used for the identification of cancer volatile organic compounds as biomarkers.
A biomarker can be a metabolite, coming from a metabolic pathway or cell process, which might be employed in the diagnostic of diseases, predict patient response towards chemical therapies and/or monitor disease recurrences. Biomarkers, e.g. aldehydes or hydrocarbons, are often identified from different body fluids such as blood, urine, serum, saliva or from various tissues samples, and their concentration can vary from one sample to the other. However, the detection and the action of these biomarkers for diseases is a complicated process. Cancer is one of the main cause of death worldwide. The main characteristic of cancerous tumor is the uncontrolled growing of cells inside the organism. Likely, these uncontrolled growths are as consequence changes in the metabolism that could be analytically monitored. Depending on where the cancer cells are located, they provide different characteristics profiles. These profiles as fingerprints are used for differentiation in a comparison to normal cells. This critical study aimed at highlighting the latest progress in this area, especially in the employment of gas chromatography for the monitoring of volatile organic compounds (VOCs) and the identification of possible molecules used as biomarkers for cancer therapy.


Efficiency analysis of the MARS2013 planning strategy.
On future human Mars missions, surface sojourn duration and resources will be limited. One possibility to approach a positive cost-benefit ratio of a human Mars mission will be to maximize the scientific research on the martian surface within the operational, environmental, and experimental limitations. Sophisticated mission operations and advanced planning strategies will be a basic requirement for Mars exploration and make operational methods an important research subject during current Mars analog missions conducted by the Austrian Space Forum. The Morocco Mars analog field simulation MARS2013, with a duration of 4 weeks and a simulated time delay of 10 min for communication between field and mission support, was the most advanced Mars analog mission conducted by the Austrian Space Forum so far. Due to these constraints, planning strategies based on real-time adjustments and ground control were not applicable, leading to the development of the "3-days-in-advance" planning strategy. Applying this methodology, 45.7% of the total EVA time was used for scientific experiments that satisfied 75.8% of the planned science goals. Taking into account all activities (donning of the suits included), the fraction of time used for scientific experiments made up only one-third of the total time, whereas it was planned to be above 50%. Here, we present details on the application of the 3-days-in-advance planning and on the results of the efficiency analysis of MARS2013.


Mechanisms of active H+ secretion in the proximal tubule.
This Editorial Review describes the mechanisms that have been identified for mediating active H+ secretion across the luminal membrane of the proximal tubular cell. These include Na+-H+ exchange, Na+-phosphate cotransport, organic anion-OH- exchange, ATP-driven H+ transport, and redox-driven H+ transport. Of these, Na+-H+ exchange appears to be the most important. Indeed, there is no conclusive evidence against the concept that luminal membrane Na+-H+ exchange is the principal if not the sole mechanism for mediating active H+ secretion in the proximal tubule. The transport properties of the Na+-H+ exchanger, particularly its striking sensitivity to intracellular pH, can explain many aspects of how the rate of active H+ secretion in the proximal tubule is regulated in response to various physiologic stimuli. Certainly, important physiologic roles for other H+ secretory mechanisms, especially those involving primary active H+ secretion, have not been excluded. Nevertheless, there is not yet definitive proof that ATP-driven or redox-driven H+ pumps are located on the luminal membrane itself or that they significantly contribute to transtubular H+ secretion.


Participant demographics reported in "Table 1" of randomised controlled trials: a case of "inverse evidence"?
Data supporting external validity of trial results allows clinicians to assess the applicability of a study's findings to their practice population. Socio-economic status (SES) of trial participants may be critical to external validity given the relationship between social and economic circumstances and health. We explored how this is documented in reports of RCTs in four major general medical journals. The contents lists of four leading general medical journals were hand searched to identify 25 consecutive papers reporting RCT results in each journal (n = 100). Data on demographic characteristics were extracted from each paper's Table 1 only (or equivalent). Authors infrequently reported key demographic characteristics relating to SES of RCT participants. Age and gender of participants were commonly reported. Less than 10% reported occupational group, employment status, income or area based measures of disadvantage. Without adequate reporting of key indicators of SES in trial participants it is unclear if lower SES groups are under-represented. If such groups are systematically under-recruited into trials, this may limit the external validity and applicability of study findings to these groups. This is in spite of the higher health-care need in more disadvantaged populations. Under-representation of low SES groups could underestimate the reported effect of an intervention for those with a higher baseline risk. The marginal benefit identified in a trial with poor or no representation of lower SES participants could significantly underestimate the potential benefit to a low SES community. More transparency in this reporting and greater attention to the impact of SES on intervention outcomes in clinical trials is needed. This could be considered in the next revision of the CONSORT statement.


Controlling Near-Surface Ni Composition in Octahedral PtNi(Mo) Nanoparticles by Mo Doping for a Highly Active Oxygen Reduction Reaction Catalyst.
We report and study the translation of exceptionally high catalytic oxygen electroreduction activities of molybdenum-doped octahedrally shaped PtNi(Mo) nanoparticles from conventional thin-film rotating disk electrode screenings (3.43 ± 0.35 A mgPt-1 at 0.9 VRHE) to membrane electrode assembly (MEA)-based single fuel cell tests with sustained Pt mass activities of 0.45 A mgPt-1 at 0.9 Vcell, one of the highest ever reported performances for advanced shaped Pt alloys in real devices. Scanning transmission electron microscopy with energy dispersive X-ray analysis (STEM-EDX) reveals that Mo preferentially occupies the Pt-rich edges and vertices of the element-anisotropic octahedral PtNi particles. Furthermore, by combining in situ wide-angle X-ray spectroscopy, X-ray fluorescence, and STEM-EDX elemental mapping with electrochemical measurements, we finally succeeded to realize high Ni retention in activated PtNiMo nanoparticles even after prolonged potential-cycling stability tests. Stability losses at the anodic potential limits were mainly attributed to the loss of the octahedral particle shape. Extending the anodic potential limits of the tests to the Pt oxidation region induced detectable Ni losses and structural changes. Our study shows on an atomic level how Mo adatoms on the surface impact the Ni surface composition, which, in turn, gives rise to the exceptionally high experimental catalytic ORR reactivity and calls for strategies on how to preserve this particular surface composition to arrive at performance stabilities comparable with state-of-the-art spherical dealloyed Pt core-shell catalysts.


The chromosomal virulence gene, chvE, of Agrobacterium tumefaciens is regulated by a LysR family member.
Certain plant phenolic compounds and monosaccharides induce the transcription of virulence (vir) genes of Agrobacterium tumefaciens through the VirA-VirG two-component regulatory system. The product of the chromosomal virulence gene chvE is homologous to galactose-binding protein of Escherichia coli and is required for vir gene induction by sugars. Adjacent to, but divergent in transcription from, chvE is an open reading frame, now termed gbpR (galactose-binding protein regulator), that is homologous to the LysR family of transcriptional regulators. chvE::lacZ expression was induced by L-arabinose, D-galactose, and D-fucose when gbpR was present. In the absence of inducer, GbpR repressed chvE::lacZ expression. In addition, GbpR negatively regulated its own expression.


Simple intraoperative technique for hemostasis of cervical venous bleeding.
To present a simple, fast, and efficient technique to control bleeding from the epidural venous plexus during anterior cervical discectomy. When encountering persistent bleeding from the venous epidural plexus during anterior cervical discectomy that cannot be controlled by bipolar coagulation, one can take approximately 5 to 10 ml of saline and connect the syringe to a curved blunt cannula with an atraumatic tip. Subsequently, the atraumatic tip of the syringe is placed between the dura and the bony edge and then the saline is powerfully injected into the epidural space. This will stop the bleeding. This technique has been applied in 118 cases of anterior cervical discectomies and has proven to be very effective. We did not have to apply another hemostatic technique to control bleeding from the epidural plexus in any of our cases. We present a simple and effective hemostasis technique to control bleeding from the cervical epidural venous plexus by injecting saline into the epidural space. We speculate that the mechanism by which the bleeding is controlled consists of filling the epidural space with saline; this elevated "pressure" controls the bleeding in a very effective way. Moreover, some venous contraction may occur because of the saline injection.


Fracture liaison services for the evaluation and management of patients with osteoporotic fracture: a cost-effectiveness evaluation based on data collected over 8 years of service provision.
The cost-effectiveness of Fracture Liaison Services (FLSs) for prevention of secondary fracture in osteoporosis patients in the United Kingdom (UK), and the cost associated with their widespread adoption, were evaluated. An estimated 18 fractures were prevented and £21,000 saved per 1,000 patients. Setup across the UK would cost an estimated £9.7 million. Only 11% to 28% of patients with a fragility fracture receive osteoporosis treatment in the UK. FLSs provide an efficient means to identify patients and are endorsed by the Department of Health but have not been widely adopted. The objective of this study was to evaluate the cost-effectiveness of FLSs in the UK and the cost associated with their widespread adoption. A cost-effectiveness and budget-impact model was developed, utilising detailed audit data collected by the West Glasgow FLS. For a hypothetical cohort of 1,000 fragility-fracture patients (740 requiring treatment), 686 received treatment in the FLS compared with 193 in usual care. Assessments and osteoporosis treatments cost an additional £83,598 and £206,544, respectively, in the FLS; 18 fractures (including 11 hip fractures) were prevented, giving an overall saving of £21,000. Setup costs for widespread adoption of FLSs across the UK were estimated at £9.7 million. FLSs are cost-effective for the prevention of further fractures in fragility-fracture patients. The cost of widespread adoption of FLS across the UK is small in comparison with other service provision and would be expected to result in important benefits in fractures avoided and reduced hospital bed occupancy.


Is there an association between eating behaviour and attention-deficit/hyperactivity disorder symptoms in preschool children?
There is some evidence that aberrant eating behaviours and obesity co-occur with attention-deficit/hyperactivity disorder (ADHD) symptoms. The present study is the first that aims to investigate the association between eating behaviours and ADHD symptoms in early childhood in a population-based cohort. We included 471 preschool children from the Rhea mother-child cohort in Crete, Greece. Parents completed the Children's Eating Behaviour Questionnaire to assess children's eating behaviour and the 36-item ADHD test (ADHDT) to evaluate ADHD symptoms at 4 years of age. Multivariable linear regression models were used to examine the association of eating behaviours with ADHD symptoms. Regarding children's food approach eating behaviours, we observed a positive association between food responsiveness and total ADHD index, as well as impulsivity, inattention and hyperactivity subscale, separately. Similarly, there was a significant positive association between emotional overeating and ADHD symptoms. With regard to children's food avoidant behaviours, food fussiness was found to be significantly associated with the impulsivity subscale. A dose-response association between the food approach behaviours and ADHD symptoms was also observed. Children on the medium and highest tertile of the food responsiveness subscale had increased scores on the ADHD total scale, as compared to those on the lowest tertile. As regards emotional overeating, children in the highest tertile of the scale had higher scores on ADHD total and hyperactivity. Our findings provide evidence that food approach eating behaviours such as food responsiveness and emotional overeating are associated with the increased ADHD symptoms in preschool children. Future studies to better understand this overlap will enhance potential interventions.


How do women prepare for pregnancy? Preconception experiences of women attending antenatal services and views of health professionals.
To determine the extent to which women plan and prepare for pregnancy. Cross-sectional questionnaire survey of pregnant women attending three maternity services in London about knowledge and uptake of preconception care; including a robust measure of pregnancy planning, and phone interviews with a range of health care professionals. We recruited 1173/1288 (90%) women, median age of 32 years. 73% had clearly planned their pregnancy, 24% were ambivalent and only 3% of pregnancies were unplanned. 51% of all women and 63% of those with a planned pregnancy took folic acid before pregnancy. 21% of all women reported smoking and 61% reported drinking alcohol in the 3 months before pregnancy; 48% of smokers and 41% of drinkers reduced or stopped before pregnancy. The 51% of all women who reported advice from a health professional before becoming pregnant were more likely to adopt healthier behaviours before pregnancy [adjusted odds ratios for greatest health professional input compared with none were 2.34 (95% confidence interval 1.54-3.54) for taking folic acid and 2.18 (95% CI 1.42-3.36) for adopting a healthier diet before pregnancy]. Interviews with 20 health professionals indicated low awareness of preconception health issues, missed opportunities and confusion about responsibility for delivery of preconception care. Despite a high level of pregnancy planning, awareness of preconception health among women and health professionals is low, and responsibility for providing preconception care is unclear. However, many women are motivated to adopt healthier behaviours in the preconception period, as indicated by halving of reported smoking rates in this study. The link between health professional input and healthy behaviour change before pregnancy is a new finding that should invigorate strategies to improve awareness and uptake of pre-pregnancy health care, and bring wider benefits for public health.


Light colour and intensity alters reproductive/seasonal responses in Japanese quail.
An extensive literature is available on the photoperiodic responses of avian species but studies on light colour and wavelength from light emitting diode (LED) sources on reproduction are limited. Hence, an experiment was designed to study the effect of different colours and intensities of light on the reproductive responses of Japanese quail. Three-week old quail were exposed to five different light conditions with a long photoperiod (LD 16:8): WT (white fluorescent light 100 lux as control), W LED (white light emitting diode, 30 lux), B LED (blue LED, 30 lux), G LED (green LED, 30 lux) and R-LED (red LED, 30 lux). The cloacal gland size, an indicator of androgenic activity, was monitored weekly. The results indicated an early initiation of gonadal growth in WT quail which continued and maintained a plateau throughout the period of study. On the other hand, in general low intensity light, there was a decreased amplitude of the reproductive cycle and the quail exposed to different colour lights (green, red and blue lights) used different incubation times to initiate their gonadal growth and exhibited a gonadal cycle of a different duration up to 15.5 weeks. Thereafter, the gonad of quail of all the LED groups started developing again (including the blue LED exposed quail which remained undeveloped until this age) and attained the increased degree of growth until 26.5 weeks of age. During the second cycle, gonads of green and red light exposed quail continued to increase and maintained a plateau of development similar to WT exposed control while white and blue LED exposed quail exhibited spontaneous regression and attained complete sexual quiescence. Based on our study, it is suggested that long term exposure to blue LED light of low intensity may induce gonadal regression even under long-day conditions (LD 16:8), while exposure to green and red lights appears to maintain a constant photosensitivity after one complete gonadal cycle.


The effects of probiotic supplement on hemoglobin in chronic renal failure patients under hemodialysis: A randomized clinical trial.
Chronic inflammation is one of the causes of anemia in chronic renal failure patients under hemodialysis. Probiotics probably establish a balance between pro- and anti-inflammatory cytokines. The study was conducted to determine the effects of probiotic supplementation on hemoglobin (Hb) in hemodialysis patients. A parallel clinical trial was conducted in which patients were randomly allocated into two groups. The intervention group (n = 18) was given a 500 mg probiotic supplement (a capsule) every day whereas the control group (n = 18) received placebo (a capsule), both for 3 months. Hb levels and C-reactive protein (CRP) levels were measured for three periods. The data were analyzed in SPSS-16 using statistical tests including the t-test and repeated-measures ANOVA. In the probiotic supplementation group, the mean Hb was 9.22 ± 1.04 mg/dl before the intervention and reached 10.85 ± 1.177 mg/dl afterward, while in the placebo group, the mean Hb level was 9.38 ± 0.97 mg/dl before the intervention and reached 10.03 ± 1.97 mg/dl afterward (P > 0.05). During the study, the placebo caused to increase of Hb temporary, but in longer term, the effect of probiotic was more manifested. Hb levels increased in both groups although the change was not statistically significant (P > 0.05). The findings showed no significant differences between the two groups in either the pre- or post-intervention CRP levels (P = 0.239). Probiotic supplementation decreased Hb fluctuations in hemodialysis patients but did not result in a significant increase in Hb levels. Similar studies are therefore recommended to be conducted with a prolonged duration of the study or an increased probiotic dose with larger sample size to complete the results of the present study.


Biodegradable elastomers and silicon nanomembranes/nanoribbons for stretchable, transient electronics, and biosensors.
Transient electronics represents an emerging class of technology that exploits materials and/or device constructs that are capable of physically disappearing or disintegrating in a controlled manner at programmed rates or times. Inorganic semiconductor nanomaterials such as silicon nanomembranes/nanoribbons provide attractive choices for active elements in transistors, diodes and other essential components of overall systems that dissolve completely by hydrolysis in biofluids or groundwater. We describe here materials, mechanics, and design layouts to achieve this type of technology in stretchable configurations with biodegradable elastomers for substrate/encapsulation layers. Experimental and theoretical results illuminate the mechanical properties under large strain deformation. Circuit characterization of complementary metal-oxide-semiconductor inverters and individual transistors under various levels of applied loads validates the design strategies. Examples of biosensors demonstrate possibilities for stretchable, transient devices in biomedical applications.


Adenosine pre- and postsynaptic modulation of glutamate-dependent calcium activity in hypothalamic neurons.
1. Within the hypothalamus, adenosine has been reported to influence temperature regulation, sleep homeostasis, and endocrine secretions. The effects of adenosine on hypothalamic neurons have not been studied at the cellular level. Adenosine (5 nM-30 microM) showed no influence on intracellular Ca2+ or electrical activity in the presence of glutamate receptor antagonists D-2-amino-5-phosphonovalerate and 6-cyano-7-nitroquinoxaline-2,3-dione; consequently, we examined the role of adenosine in modulating the activity of glutamate in cultured hypothalamic neurons (n > 1,700) with fura-2 Ca2+ digital imaging and whole cell patch-clamp electrophysiology in the absence of glutamate receptor block. 2. When glutamate receptors were not blocked, adenosine (1-30 microM) and the selective adenosine A1 receptor agonist N6-cyclopentyl adenosine (CPA; 5 nM-1 microM) caused a large reduction in intracellular Ca2+ and electrical activity, suggesting that glutamate neurotransmission was critical for an effect of adenosine to be detected. Neuronal Ca2+ levels were reversibly depressed by CPA (50 nM), with a maximum depression of 90%, and these effects were blocked by coadministration of the A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). 3. Ca2+ levels in immature neurons before the time of synaptogenesis were not affected by adenosine. Adenosine A1 receptor activation suppressed glutamate-mediated Ca2+ activity in neurons in vitro 8 to 73 days. 4. Adenosine (1 or 10 microM) caused a hyperpolarization of membrane potential and a reduction of large postsynaptic potentials arising from endogenously released glutamate. The administration of low concentrations of CPA (5 nM) decreased the frequency of glutamate-mediated, neuronally synchronized Ca2+ transients and the frequency of postsynaptic potentials. 5. To compare the relative effects of adenosine on hypothalamic neurons with cells from other brain regions, we assayed the effects of CPA on glutamate-mediated Ca2+ in hippocampal and cortical cultures. CPA (50 nM) reversibly depressed glutamate-mediated Ca2+ rises in hypothalamic neurons by 35%, compared with 54% in hippocampal neurons and 46% in cortical neurons. 6. If it does play a functional role, adenosine should be released by hypothalamic cells. In some neurons the adenosine A1 receptor antagonists cyclopentyltheophylline or DPCPX caused an increase in intracellular Ca2+, suggesting that adenosine was secreted by hypothalamic cells, tonically depressing glutamate-enhanced neuronal Ca2+. 7. To determine whether adenosine could exert a postsynaptic effect, we coapplied it with glutamate agonists in the presence of tetrodotoxin. Within subpopulations of hypothalamic neurons, adenosine and CPA either inhibited (18% of total neurons) or potentiated (6% of total neurons) responses to glutamate, N-methyl-D-aspartate, and kainate by > or = 20%. 8. In contrast to the modest effects found in neurons, responses of hypothalamic astrocytes to the application of glutamate or the metabotropic glutamate receptor agonist (+/-)-trans-1-amino-1,3-cyclopentanedicarboxylic acid were strongly potentiated by adenosine (mean +225%) and CPA. 9. Together, these findings suggest that adenosine exerts a major presynaptic effect and a minor postsynaptic effect in the modulation of glutamate neurotransmission in the hypothalamus, where it can play a significant role in blocking a large part of the glutamate-induced Ca2+ rise. In the absence of glutamate transmission, adenosine has relatively little effect on either neuronal intracellular Ca2+ or electrical activity.


Ursolic acid reverses liver fibrosis by inhibiting NOX4/NLRP3 inflammasome pathways and bacterial dysbiosis.
Activation of the NOX4/NLRP3 inflammasome pathway has been associated with fibrosis in other organs. An imbalance in intestinal bacteria is an important driving factor of liver fibrosis through the liver-gut axis. This study aimed to explore whether the effect of ursolic acid (UA) on liver fibrosis was associated with the NOX4/NLRP3 inflammasome pathways and intestinal bacteria. Wild-type (WT), NLRP3-/-, and NOX4-/- mice and AP-treated mice were injected with CCI4 and treated with or without UA. The intestinal contents of the mice were collected and analyzed by 16S rRNA sequencing. UA alleviated liver fibrosis, which manifested as decreases in collagen deposition, liver injury, and the expression of fibrosis-related factors, and the expression of NOX4 and NLRP3 was significantly inhibited by UA treatment. Even after CCI4 injection, liver damage and fibrosis-related factors were significantly decreased in NLRP3-/-, NOX4-/-, and AP-treated mice. Importantly, the expression of NLRP3 was obviously inhibited in NOX4-/- and AP-treated mice. In addition, the diversity of intestinal bacteria and the abundance of probiotics in NLRP3-/- and NOX4-/- mice was significantly higher than those in WT mice, while the abundance of harmful bacteria in NLRP3-/- and NOX4-/- mice was significantly lower than that in WT mice. The NOX4/NLRP3 inflammasome pathway plays a crucial role in liver fibrosis and is closely associated with the beneficial effect of UA. The mechanism by which the NOX4/NLRP3 inflammasome pathway is involved in liver fibrosis may be associated with disordered intestinal bacteria.


Effects of altitude on exercise level and heart rate in patients with coronary artery disease and healthy controls.
Background. To evaluate the safety and effects of high altitude on exercise level and heart rate in patients with coronary artery disease compared with healthy controls.Methods. Eight patients with a history of an acute myocardial infarction (ejection fraction >5%) with a low-risk score were compared with seven healthy subjects during the Dutch Heart Expedition at the Aconcagua in Argentina in March 2007. All subjects underwent a maximum exercise test with a cycle ergometer at sea level and base camp, after ten days of acclimatisation, at an altitude of 4200 m. Exercise capacity and maximum heart rate were compared between groups and within subjects.Results. There was a significant decrease in maximum heart rate at high altitude compared with sea level in both the patient and the control group (166 vs. 139 beats/min, p<0.001 and 181 vs. 150 beats/min, p<0.001). There was no significant difference in the decrease of the exercise level and maximum heart rate between patients and healthy controls (-31 vs. -30%, p=0.673).Conclusion. Both patients and healthy controls showed a similar decrease in exercise capacity and maximum heart rate at 4200 m compared with sea level, suggesting that patients with a history of coronary artery disease may tolerate stay and exercise at high altitude similarly to healthy controls. (Neth Heart J 2010;18:118-21.).


Comparative effectiveness of rhythm control vs rate control drug treatment effect on mortality in patients with atrial fibrillation.
Controversy continues concerning the choice of rhythm control vs rate control treatment strategies for atrial fibrillation (AF). A recent clinical trial showed no difference in 5-year mortality between the 2 treatments. We aimed to determine whether the 2 strategies have similar effectiveness when applied to a general population of patients with AF with longer follow-up. We used population-based administrative databases from Quebec, Canada, from 1999 to 2007 to select patients 66 years or older hospitalized with an AF diagnosis who did not have AF-related drug prescriptions in the year before the admission but received a prescription within 7 days of discharge. Patients were followed until death or administrative censoring. Mortality was analyzed by multivariable Cox regression. Among 26,130 patients followed for a mean (SD) period of 3.1 years (2.3 years), there were 13,237 deaths (49.5%). After adjusting for covariates, we found that the effect of rhythm vs rate control drugs changed over time: after a small increase in mortality for patients treated with rhythm control in the 6 months following treatment initiation (hazard ratio [HR], 1.07; 95% CI, 1.01-1.14), the mortality was similar between the 2 groups until year 4 but decreased steadily in the rhythm control group after year 5 (HR, 0.89; 95% CI, 0.81-0.96; and HR, 0.77; 95% CI, 0.62-0.95, after 5 and 8 years, respectively). In this population-based sample of patients with AF, we found little difference in mortality within 4 years of treatment initiation between patients with AF initiating rhythm control therapy vs those initiating rate control therapy. However, rhythm control therapy seems to be superior in the long-term.


UV-induced retinal proteome changes in the rat model of age-related macular degeneration.
Age-related macular degeneration (AMD) is characterized by irreversible damage of photoreceptors in the central posterior part of the retina, called the macula and is the most common cause of vision loss in those aged over 50. A growing body of evidence shows that cumulative long-term exposure to UV radiation may be harmful to the retina and possibly leads to AMD irrespective of age. In spite of many research efforts, cellular and molecular mechanisms leading to UV-induced retinal damage and possibly retinal diseases such as AMD are not completely understood. In the present study we explored damage mechanisms accounting for UV-induced retinal phototoxicity in the rats exposed to UVA and UVB irradiation using a proteomics approach. Our study showed that UV irradiation induces profound changes in the retinal proteomes of the rats associated with the disruption of energy homeostasis, oxidative stress, DNA damage response and structural and functional impairments of the interphotoreceptor matrix components and their cell surface receptors such as galectins. Two small leucine-rich proteoglycans, biglycan and lumican, were identified as phototoxicity biomarkers associated with UV-induced disruption of interphotoreceptor matrix (IPM). In addition, UVB induced activation of Src kinase, which could account for cytoskeletal rearrangements in the retina was observed at the proteomics level. Pharmacological intervention either to target Src kinase with the aim of preventing cytoskeletal rearrangements in the retinal pigment epithelium (RPE) and neuronal retina or to help rebuild damaged IPM may provide fresh avenues of treatment for patients suffering from AMD.


The association of dietary phosphorus with blood pressure: results from a secondary analysis of the PREMIER trial.
Inconsistent findings exist for the association between dietary phosphorus intake and blood pressure (BP). We examined the longitudinal association between urinary excretion and dietary intake of phosphorus (total, plant, animal, and added) with BP. This is a secondary analysis of PREMIER, a randomized behavioral intervention study in adults (25-79 years) with BP, measured at 6 months, as the primary outcome. We classified total phosphorus intake from dietary recalls into plant, animal, and added phosphorus. We modeled 6-month change of phosphorus intake (from 24 h dietary recalls, N = 622) and excretion (from 24 h urine collection, N = 564) on BP, using linear regression crude and adjusted for intervention, age, race, sex, income, education, study site, and change in energy intake (kcal/day), sodium intake (mg/day), fitness (heart rate, bpm), and DASH diet index. Baseline phosphorus intake was 1154 mg/day (95% CI 1126, 1182) with 38%, 53%, and 10% from plant, animal, and added phosphorus, respectively. Total phosphorus intake was not associated with significant changes in BP. Increased urinary phosphorus excretion was associated with a significant increase in DBP [0.14 mmHg/100 mg (0.01, 0.28), adjusted]. In several analyses, phosphorus type (plant, animal, or added) significantly modified the association between phosphorus intake and BP. For example, added phosphorus (but not plant or animal) was associated with increases in SBP and DBP, 1.24 mmHg/100 mg (0.36, 2.12) and 0.83 mmHg/100 mg (0.22, 1.44), respectively, crude. These findings suggest that the type of phosphorus may modify the association between phosphorus intake and BP. Trial registration NCT00000616 (clinicaltrials.gov).


Anti-schistosomal activities of quinoxaline-containing compounds: From hit identification to lead optimisation.
Schistosomiasis is a neglected disease of poverty that is caused by infection with blood fluke species contained within the genus Schistosoma. For the last 40 years, control of schistosomiasis in endemic regions has predominantly been facilitated by administration of a single drug, praziquantel. Due to limitations in this mono-chemotherapeutic approach for sustaining schistosomiasis control into the future, alternative anti-schistosomal compounds are increasingly being sought by the drug discovery community. Herein, we describe a multi-pronged, integrated strategy that led to the identification and further exploration of the quinoxaline core as a promising anti-schistosomal scaffold. Firstly, phenotypic screening of commercially available small molecules resulted in the identification of a moderately active hit compound against Schistosoma mansoni (1, EC50 = 4.59 μM on schistosomula). Secondary exploration of the chemical space around compound 1 led to the identification of a quinoxaline-core containing, non-genotoxic lead (compound 22). Compound 22 demonstrated substantially improved activities on both intra-mammalian (EC50 = 0.44 μM, 0.20 μM and 84.7 nM, on schistosomula, juvenile and adult worms, respectively) and intra-molluscan (sporocyst) S. mansoni lifecycle stages. Further medicinal chemistry optimisation of compound 22, resulting in the generation of 20 additional analogues, improved our understanding of the structure-activity relationship and resulted in considerable improvements in both anti-schistosome potency and selectivity (e.g. compound 30; EC50 = 2.59 nM on adult worms; selectivity index compared to the HepG2 cell line = 348). Some derivatives of compound 22 (e.g. 31 and 33) also demonstrated significant activity against the two other medically important species, Schistosoma haematobium and Schistosoma japonicum. Further optimisation of this class of anti-schistosomal is ongoing and could lead to the development of an urgently needed alternative to praziquantel for assisting in schistosomiasis elimination strategies.


Surgery for drug-resistant tuberous sclerosis complex-associated epilepsy: who, when, and what.
Tuberous sclerosis complex (TSC) is a multisystem genetic disorder associated with refractory early-onset epilepsy. Current evidence supports surgery as the intervention most likely to achieve long-term seizure freedom, but no specific guidelines are available on TSC pre-surgical workup. This critical review assesses which TSC patients are suitable for surgical treatment, when pre-surgical evaluation should start, and what degree of surgical resection is optimal for postsurgical outcome. We searched for publications from 2000 to 2020 in Pubmed and Embase using the terms "tuberous sclerosis," "epilepsy," and "epilepsy surgery". To evaluate postsurgical seizure outcome, we selected only studies with at least one year of follow-up. Overall, we collected data on 1,026 patients from 34 studies. Age at surgery ranged from one month to 54 years. Mean age at surgery was 8.41 years. Of the diagnostic non-invasive pre-surgical tools, MRI and video-EEG were considered most appropriate. Promising data for epileptogenic tuber detection is provided from invasive SEEG studies. Data on surgery and related outcome were available for 769 patients. Seizure freedom was seen in 64.4% of patients who underwent tuberectomy, 68.9% treated with lobectomy and 65.1% with multilobar resection. The most effective surgical approach was lobectomy, even though more recently tuberectomy associated with the resection of the perituberal area seems to be the best approach to reach seizure freedom. Published postsurgical seizure freedom rates in patients with TSC were between 65% and 75%, but reduced to 48%-57% over longer follow-up periods. Early surgery might positively affect neurodevelopmental trajectory in some patients, even though data on cognitive outcome are still to be confirmed with longitudinal studies. Considering the strong correlation between epilepsy duration and neurocognitive outcome, all patients with TSC ought to be referred early to a dedicated epilepsy centre for individually tailored pre-surgical evaluation by a multi-disciplinary epilepsy surgery team.


An experimentally informed evolutionary model improves phylogenetic fit to divergent lactamase homologs.
Phylogenetic analyses of molecular data require a quantitative model for how sequences evolve. Traditionally, the details of the site-specific selection that governs sequence evolution are not known a priori, making it challenging to create evolutionary models that adequately capture the heterogeneity of selection at different sites. However, recent advances in high-throughput experiments have made it possible to quantify the effects of all single mutations on gene function. I have previously shown that such high-throughput experiments can be combined with knowledge of underlying mutation rates to create a parameter-free evolutionary model that describes the phylogeny of influenza nucleoprotein far better than commonly used existing models. Here, I extend this work by showing that published experimental data on TEM-1 beta-lactamase (Firnberg E, Labonte JW, Gray JJ, Ostermeier M. 2014. A comprehensive, high-resolution map of a gene's fitness landscape. Mol Biol Evol. 31:1581-1592) can be combined with a few mutation rate parameters to create an evolutionary model that describes beta-lactamase phylogenies much better than most common existing models. This experimentally informed evolutionary model is superior even for homologs that are substantially diverged (about 35% divergence at the protein level) from the TEM-1 parent that was the subject of the experimental study. These results suggest that experimental measurements can inform phylogenetic evolutionary models that are applicable to homologs that span a substantial range of sequence divergence.


The beneficial effects of tadalafil on renal ischemia-reperfusion injury in rats.
Acute renal failure due to ischemia-reperfusion (I/R) injury is a common complication in cardiovascular surgery. We determined the influence of tadalafil on renal injury in a renal I/R model in rats. For this purpose, 21 male Wistar albino rats were separated into 3 groups: sham, placebo and tadalafil. A right nephrectomy was performed, and the left renal pedicles were occluded for 60 min and reperfused for 60 min in the placebo and tadalafil groups. A single dose of tadalafil (10 mg/kg) through an orogastric tube was administered to the tadalafil group. Tubular atrophy with acute inflammation in renal histology, total oxidant status (TOS) and total antioxidant status (TAS) were determined in tissue homogenates. Compared to the tadalafil group, tubular atrophy and acute inflammation was significant in the placebo group. TAS levels were significantly higher in the tadalafil group compared to the placebo (p = 0.01) and sham groups (p = 0.04). While TOS levels were significantly higher in the placebo group (p = 0.03), tadalafil did not significantly alter the TOS levels. The beneficial effects of tadalafil can be attributed to its protective effects on renal tubular cells and inhibition of leukocyte infiltration in renal tissue. We think that tadalafil treatment has an important role in reducing renal injury resulting from renal I/R.


Morbidity and mortality in hemodialysis patients.
Increase in mortality in hemodialysis patients in the United States is a rising concern. In this study, the annual mortality rate from 1981 to 1989 was determined and the morbidity and mortality of 136 patients dialyzed during 1988 was analyzed. All patients were followed with urea kinetics (UK). Patients were divided according to NCDS mapping of BUN and protein catabolic rate (pcr): Group 1, inadequate protein intake (n = 28); Group 2, adequate dialysis (n = 51); Group 3, excessive dialysis (n = 28); Group 4, undefined domain (n = 13); and Group 5, transitional domain (n = 16). The UK parameters measured were midweek predialysis BUN, pcr, and Kt/V. Group 1 by definition had the lowest pcr (0.7 g/kg/day vs. greater than 1.0 in the other groups). Patients in Group 1 had significantly lower BUN, creatinine, hematocrit, and serum albumin. Results of all other laboratory tests were not significantly different. Numbers of hospitalizations and days in the hospital were higher in Group 1. Causes of hospitalization were similar in all groups. Mortality rate was 8% in Group 1 vs. 4.4% in Group 2. Dialysis prescription using UK has maintained the annual mortality rate at about 14% for the past nine years.


Paratope Duality and Gullying are Among the Atypical Recognition Mechanisms Used by a Trio of Nanobodies to Differentiate Ebolavirus Nucleoproteins.
We had previously shown that three anti-Marburg virus nanobodies (VHH or single-domain antibody [sdAb]) targeted a cryptotope within an alpha-helical assembly at the nucleoprotein (NP) C-terminus that was conserved through half a century of viral evolution. Here, we wished to determine whether an anti-Ebola virus sdAb, that was cross-reactive within the Ebolavirus genus, recognized a similar structural feature upstream of the ebolavirus NP C-terminus. In addition, we sought to determine whether the specificities of a less cross-reactive anti-Zaire ebolavirus sdAb and a totally specific anti-Sudan ebolavirus sdAb were the result of exclusion from this region. Binding and X-ray crystallographic studies revealed that the primary determinant of cross-reactivity did indeed appear to be a preference for the helical feature. Specificity, in the case of the Zaire ebolavirus-specific sdAb, arose from the footprint shifting away from the helices to engage more variable residues. While both sdAbs used CDRs, they also had atypical side-on approaches, with framework 2 helping to accommodate parts of the epitope in sizeable paratope gullies. The Sudan ebolavirus-specific sdAb was more remarkable and appeared to bind two C-terminal domains simultaneously via nonoverlapping epitopes-"paratope duality." One mode involved paratope gullying, whereas the other involved only CDRs, with CDR3 restructuring to wedge in between opposing walls of an interdomain crevice. The varied routes used by sdAbs to engage antigens discovered here deepen our appreciation of the small scaffold's architectural versatility and also reveal lucrative opportunities within the ebolavirus NP C-termini that might be leveraged for diagnostics and novel therapeutic targeting.


Hepatitis G virus (HGV) infection & its pathogenic significance in patients of cirrhosis.
In the present study the hepatitis G virus (HGV) infection and its pathogenic significance in patients of cirrhosis were assessed using reverse transcription plus nested polymerase chain reaction (RT-PCR). Serum samples were collected from a total of 50 patients of histologically proven non-alcoholic cirrhosis and from a control group consisting of 50 healthy voluntary blood donors. HGV RNA was detected by RT-PCR using primer sequences located in the conserved NS3 helicase region of HGV genome. Serological evaluation for markers of chronic infection with HBV (HBsAg, IgG anti-HBc, HBeAg) and HCV (anti-HCV) was carried out using commercially available kits. HBV DNA and HCV RNA were also tested by PCR in those samples that were found to be non-B, non-C by serological assays. Serological evidence of exposure to HBV was found in 31 (62%) and to HCV in 15 (30%) patients. HGV RNA was detected in 6 (12%) cirrhosis patients and in 2 (4%) healthy blood donors but the difference between the two groups was not statistically significant. Of the 6 HGV positive patients, 2 were coinfected with HBV, 1 with HCV, while the remaining 3 belonged to non-B, non-C category. No significant difference was observed in the clinical and biochemical profiles of HGV-positive and HGV-negative patients except that a history of blood transfusion was significantly (P < 0.005) more common in the former. The findings indicate that the HGV infection is commonly observed in both cirrhosis patients as well as healthy blood donors. A significant association of the virus with blood transfusion is indicative of a parenteral route of transmission. The observations of this study also suggest that the pathogenic role of HGV in the causation of liver disease may be insignificant.


Intense reversal of bone marrow angiogenesis after sequential fludarabine-induction and alemtuzumab-consolidation therapy in advanced chronic lymphocytic leukemia.
Increased bone marrow (BM) angiogenesis has been demonstrated in several hematologic malignancies. BM angiogenesis is significantly decreased in patients with chronic lymphocytic leukemia (CLL) treated with fludarabine. The anti-angiogenic potential of alemtuzumab in CLL has not yet been investigated. We, therefore, evaluated BM angiogenesis in CLL patients treated sequentially with fludarabine and low doses of alemtuzumab. BM microvessel area was sequentially evaluated in 20 patients with advanced B-cell CLL who received, after a clinical response obtained with fludarabine-induction therapy, alemtuzumab, three times weekly for 6 weeks at a dose of 10 mg. The complete response rate improved from 45% after fludarabine induction to 90% after alemtuzumab consolidation. The extent of BM angiogenesis decreased continuously after either fludarabine or alemtuzumab (p=0.0002; Kruskal-Wallis test). Thirteen out of 20 (65%) patients changed from having a monoclonal to a polyclonal pattern of IgH sequences after alemtuzumab consolidation. A separate evaluation carried out in patients who achieved molecularly undetectable disease, as defined by polymerase chain reaction negativity, and in patients who remained with minimal residual disease after therapy with alemtuzumab showed a significant reduction of BM microvessel area only in the former (p=0.0002). Finally, molecular responses and a significant reduction of BM angiogenesis were more common in patients who received the cumulative planned dose of alemtuzumab (i.e.,180 mg) than in patients who received reduced doses (p=0.007 and p=0.0001, respectively). Overall, these data demonstrate a decrease in BM vascularity that characterized CLL patients who received low doses of subcutaneous alemtuzumab consolidation therapy after a clinical response to fludarabine induction therapy. Such a finding reflects, at least in part, the molecular response and cumulative dose of alemtuzumab.


Metabolic potential of a single cell belonging to one of the most abundant lineages in freshwater bacterioplankton.
Actinobacteria within the acI lineage are often numerically dominating in freshwater ecosystems, where they can account for >50% of total bacteria in the surface water. However, they remain uncultured to date. We thus set out to use single-cell genomics to gain insights into their genetic make-up, with the aim of learning about their physiology and ecological niche. A representative from the highly abundant acI-B1 group was selected for shotgun genomic sequencing. We obtained a draft genomic sequence in 75 larger contigs (sum=1.16 Mb), with an unusually low genomic G+C mol% (∼42%). Actinobacteria core gene analysis suggests an almost complete genome recovery. We found that the acI-B1 cell had a small genome, with a rather low percentage of genes having no predicted functions (∼15%) as compared with other cultured and genome-sequenced microbial species. Our metabolic reconstruction hints at a facultative aerobe microorganism with many transporters and enzymes for pentoses utilization (for example, xylose). We also found an actinorhodopsin gene that may contribute to energy conservation under unfavorable conditions. This project reveals the metabolic potential of a member of the global abundant freshwater Actinobacteria.


Viability of Lactobacillus acidophilus and Lactobacillus casei in fermented milk products during refrigerated storage.
The viability was investigated of five strains of Lactobacillus acidophilus and one strain of Lactobacillus casei that were added as adjuncts to yogurt and cultured buttermilk during 28 d of refrigerated storage at 5 to 7 degrees C. A modification of LBS (Lactobacillus selection) agar was used for the enumeration of L. acidophilus and L. casei. The medium allowed the colony formation of the adjunct bacteria while preventing colony formation of the traditional yogurt or buttermilk starter cultures. At each sampling period, colonies from the selective agar medium were isolated for confirmation of identity to confirm that only L. acidophilus and L. casei were enumerated, that their characteristics did not change during storage, or both. The strains of L. acidophilus varied in both cultured products. In buttermilk, L. acidophilus MUH-41, O-16, and L-1 exhibited no significant loss in viability, but strains 43121 and La-5 did. No significant loss in viability of L. acidophilus MUH-41 and L-1 occurred in yogurt prepared using culture CM2; however, strains 43121, O-16, and La-5 lost viability. In the yogurt prepared using culture YC-4, L. acidophilus 43121 exhibited no significant loss in viability, but MUH-41, O-16, L-1, and La-5 did. There was no loss in viability of L. casei GG during storage of any of the cultured products.


The struggle towards 'the New Normal': a qualitative insight into psychosexual adjustment to prostate cancer.
Despite the growing body of literature which highlights the potential for significant and enduring side-effects of prostate cancer treatment, there is limited research exploring the experience of living with the treatment-induced side-effects such as sexual dysfunction, and their repercussions for men and their partners. The aim of this qualitative study was to explore factors influencing psychosexual adjustment, self-perception, and unmet information and support needs of prostate cancer patients and their partners. Twenty-one men, recruited via a prostate cancer support group newsletter, participated in face-to-face semi-structured interviews, which were subjected to thematic analysis. The qualitative analysis revealed three inter-connected main themes which contributed to men's psychosexual adjustment: i) Psychosexual impact, ii) Communication and support, and iii) Integration process. Men reported distressing sexual and urinary difficulties, tainted self-perception and altered intimate relationships. Receiving adequate information and support, and having good communication with their doctors and partners facilitated better adjustment to prostate cancer treatment. Coming to terms with the significant impact of treatment had involved making lifestyle changes, coping with emotional struggles and striving to accept and integrate their post-treatment "new normal" self and sexual life. The importance of adequate communication with health professionals and partners, especially regarding treatment effects on sexual function and rehabilitation options, was highlighted as a key factor facilitating the adjustment process. Prostate cancer patients would benefit from improved access to timely and tailored information and decision-making resources, ongoing multidisciplinary care, and support groups, as well as appropriate referrals for sexual and psychological counselling.


Critical role of Bmpr1a in mandibular condyle growth.
The importance of Bone Morphogenetic Proteins (BMPs) in the regulation of cell fate, differentiation and proliferation in the growth plate is well-known. However, in secondary cartilages (such as that in the temporomandibular joint) that grow by proliferation of prechondrocytes and differ in their pattern of growth, the role of BMPs is largely unexplored. To examine this question, we ablated Bmpr1a in the condylar cartilage of neonatal mice and assessed the consequences for mandibular condyle growth and organization at intervals over the ensuing 4 weeks. Bmpr1a deficiency caused significant chondrodysplasia and almost eliminated the chondrocytic phenotype in the TMJ. Expression of Sox9, collagen II, proteoglycan were all greatly reduced, and cell proliferation as detected by BrdU was almost non-existent in the knockout mice. Primary bone spongiosa formation was also disturbed and was accompanied by reduced Osterix expression. These findings strongly suggest that Bmpr1a is critical for the development and growth of the mandibular condyle via its effect on proliferation of prechondroblasts and chondrocyte differentiation.


Dose dependent effects of nitrate supplementation on cardiovascular control and microvascular oxygenation dynamics in healthy rats.
High dose nitrate (NO3(-)) supplementation via beetroot juice (BR, 1 mmol/kg/day) lowers mean arterial blood pressure (MAP) and improves skeletal muscle blood flow and O2 delivery/utilization matching thereby raising microvascular O2 pressure (PO2mv). We tested the hypothesis that a low dose of NO3(-) supplementation, consistent with a diet containing NO3(-) rich vegetables (BRLD, 0.3 mmol/kg/day), would be sufficient to cause these effects. Male Sprague-Dawley rats were administered a low dose of NO3(-) (0.3 mmol/kg/day; n=12), a high dose (1 mmol/kg/day; BRHD, n=6) or tap water (control, n=10) for 5 days. MAP, heart rate (HR), blood flow (radiolabeled microspheres) and vascular conductance (VC) were measured during submaximal treadmill exercise (20 m/min, 5% grade, equivalent to ~60% of maximal O2 uptake). Subsequently, PO2mv (phosphorescence quenching) was measured at rest and during 180 s of electrically-induced twitch contractions (1 Hz, ~6 V) of the surgically-exposed spinotrapezius muscle. BRLD and BRHD lowered resting (control: 139 ± 4, BRLD: 124 ± 5, BRHD: 128 ± 9 mmHg, P<0.05, BRLD vs. control) and exercising (control: 138 ± 3, BRLD: 126 ± 4, BRHD: 125 ± 5 mmHg, P<0.05) MAP to a similar extent. For BRLD this effect occurred in the absence of altered exercising hindlimb muscle(s) blood flow or spinotrapezius PO2mv (rest and across the transient response at the onset of contractions, all P>0.05), each of which increased significantly for the BRHD condition (all P<0.05). Whereas BRHD slowed the PO2mv kinetics significantly (i.e., >mean response time, MRT; control: 16.6 ± 2.1, BRHD: 23.3 ± 4.7s) following the onset of contractions compared to control, in the BRLD group this effect did not reach statistical significance (BRLD: 20.9 ± 1.9s, P=0.14). These data demonstrate that while low dose NO3(-) supplementation lowers MAP during exercise it does so in the absence of augmented muscle blood flow, VC and PO2mv; all of which are elevated at a higher dose. Thus, in healthy animals, a high dose of NO3(-) supplementation seems necessary to elicit significant changes in exercising skeletal muscle O2 delivery/utilization.


Immunocytochemically defined interneuron populations in the hippocampus of mouse strains used in transgenic technology.
Transgenic mice are overtaking the role of model animals in neuroscience. They are used in developmental, anatomical, and physiological as well as experimental neurology. However, most results on the organization of the nervous system derive from the rat. The rat hippocampus and its neuronal elements have been thoroughly investigated, revealing remarkable functional and morphological diversity and specificity among hippocampal interneurons. Our aim was to examine the properties of distinct hippocampal interneuron populations, i.e., those immunoreactive for calcium-binding proteins (parvalbumin, calbindin, and calretinin), neuropeptides (cholecystokinin, neuropeptide Y, somatostatin, vasoactive intestinal polypeptide), and certain receptors (metabotropic glutamate receptor 1alpha, cannabinoid receptor type 1) in four strains of mice widely used in transgenic technology, and to compare their properties to those in the rat. Our data indicate that the distribution as well as the dendritic and axonal arborization of mouse interneurons immunoreactive for the different markers was identical in the examined mouse strains, and in most respects are similar to the features found in the rat. The postsynaptic targets of neurons terminating in the perisomatic (parvalbumin), proximal (calbindin), and distal (somatostatin) dendritic region, as well as on other interneurons (calretinin), also matched those found in the rat. However, a few significant differences could also be observed between the two species in addition to the already described immunoreactivity of mossy cells for calretinin: the absence of spiny calretinin-immunoreactive interneurons in the CA3 region, sparse contacts between calretinin-immunoreactive interneurons, and the axon staining for somatostatin and neuropil labeling for cholecystokinin. We can conclude that the morphofunctional classification of interneurons established in the rat is largely valid for mouse strains used in transgenic procedures.


Predictors of atrial fibrillation after conventional and beating heart coronary surgery: A prospective, randomized study.
Atrial fibrillation (AF) increases the morbidity of CABG. The pathophysiology is uncertain, and its prevention remains suboptimal. This prospective, randomized study was designed to define the role of cardiopulmonary bypass (CPB) and cardioplegic arrest in the pathogenesis of this complication. Two hundred patients were prospectively randomized to (1) on-pump conventional surgery [(100 patients, 79 men, mean age 63 (40 to 77) years)] with normothermic CPB and cardioplegic arrest of the heart or (2) off-pump surgery [(100 patients, 82 men, mean age 63 (38 to 86) years)] on the beating heart. Heart rate and rhythm were continuously monitored with an automated arrhythmia detector during the first 72 hours after surgery. Thereafter, routine clinical observation was performed and continuous monitoring restarted in the case of arrhythmia. The association of perioperative factors with AF was investigated by univariate analysis. Significant variables were then included into a stepwise logistic regression model to ascertain their independent influence on the occurrence of AF. There were no significant baseline differences between groups. Thirty-nine patients in the on-pump group and 8 patients in the off-pump group had postoperative sustained AF (P:=0.001). Univariate analysis showed that CPB inclusive of cardioplegic arrest, postoperative inotropic support, intubation time, chest infection, and hospital length of stay were predictors of AF (all P:<0.05). However, stepwise multivariate regression analysis identified CPB inclusive of cardioplegic arrest as the only independent predictor of postoperative AF (OR 7.4; CI 3.4 to 17.9). CPB inclusive of cardioplegic arrest is the main independent predictor of postoperative AF in patients undergoing coronary revascularization.


Knotted nasoenteric feeding tube in an infant.
This is a case report, with review of the literature, of an infant in the neonatal intensive care unit (NICU) with the complication of a nasoenteric weighted feeding tube that assumed an overhand knot configuration. Knotting of the feeding tube is a rare occurrence related to the use of feeding tubes, and should be considered if the tube becomes nonfunctional, if there are clinical factors that might lead to displacement or malposition of the tube, or if there is excessive resistance or sudden onset of respiratory distress in association with attempted removal.


Reproductive profiles of adult Mongolian gerbils gestated as the sole fetus in a uterine horn.
We compared reproductive profiles of Mongolian gerbils gestated alone in a uterine horn (Isolate males and Isolate females) with those of gerbils gestated in intrauterine positions between two male fetuses (2M males and 2M females) and two female fetuses (2F males and 2F females). We found that, when adult, the reproductive profiles of gerbils that had been gestated as isolates resembled that of gerbils that had been gestated as 2F fetuses: 1. Isolate females gestated litters containing both the same proportion of males as the litters of 2F females and a significantly smaller proportion of males than litters of 2M females. 2. Isolate males, like 2F males, were less likely to impregnate females than were 2M males, and 3. both 2F males and isolate males exhibited disturbed patterns of copulation and reduced levels of scent-marking relative to 2M males. Our results were entirely consistent with the view that intrauterine exposure to males, but not to females, was responsible for previously described differences in the reproductive profiles of 2M and 2F Mongolian gerbils.


Brief intervention in alcohol-dependent versus nondependent individuals.
Randomized trials examining the effects of brief alcohol interventions by primary care providers have consistently excluded individuals with alcohol dependence. The purpose of this study was to examine whether a diagnosis of alcohol dependence, according to the criteria in Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, predicted differential effectiveness of a brief intervention (BI). Retrospective analyses were performed on participants (N = 326) enrolled in a randomized trial designed to examine the impact of interactive voice response following BI. All participants had received a BI from their primary care provider before enrolling in the study. Daily consumption data were collected using the Timeline Followback for the period before the BI (mean = 71 days) and for 6 months following the BI. We compared nondependent and dependent participants on a number of consumption-based outcomes. Dependent participants had significantly higher pre-BI consumption. At the index assessment (median =15 days after BI), both dependent and nondependent participants reported significant reductions in total drinks per week and drinking days per week after BI. Dependent participants significantly reduced their drinks per drinking day. After BI, dependent participants no longer differed significantly from nondependent participants on these measures. Similar decreases from before BI were observed in both groups through 6 months, although dependent participants drank on fewer days and significantly more on days on which they drank than did nondependent participants. Regression analyses showed that baseline consumption was the only significant predictor of post-BI consumption. We found no evidence that dependent participants realized less benefit on measures of alcohol consumption following a BI than nondependent participants.


Large scale enzyme based xenobiotic identification for exposomics.
Advances in genomics have revealed many of the genetic underpinnings of human disease, but exposomics methods are currently inadequate to obtain a similar level of understanding of environmental contributions to human disease. Exposomics methods are limited by low abundance of xenobiotic metabolites and lack of authentic standards, which precludes identification using solely mass spectrometry-based criteria. Here, we develop and validate a method for enzymatic generation of xenobiotic metabolites for use with high-resolution mass spectrometry (HRMS) for chemical identification. Generated xenobiotic metabolites were used to confirm identities of respective metabolites in mice and human samples based upon accurate mass, retention time and co-occurrence with related xenobiotic metabolites. The results establish a generally applicable enzyme-based identification (EBI) for mass spectrometry identification of xenobiotic metabolites and could complement existing criteria for chemical identification.