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Palm wine fermentation is a complex microbial process that evolves with tapping times. The dynamics in microbiota and metabolites throughout palm wine tapping days is still not established, which are critical for the distinctive characteristics of palm wine taste and quality, and thus the mastery of the daily quality fluctuation during tapping. We analyzed the changes in microbial community structure by amplicon sequencing of bacterial 16S rRNA gene and fungal internal transcribed spacer (ITS) region, and metabolite profile changes using mass spectrometry in palm wine collected over 25–30 days tapping of ron (Borassus aethiopum) and oil palms (Elaeis guineensis) from Côte d’Ivoire. The stage-wise collected palm wine samples showed distinct changes in microbial diversity and pH, supporting microbial community dynamics during palm wine tapping. Results highlighted the dominance ofSaccharomyces cerevisiaein early stages and the emergence of non-Saccharomycesyeasts, particularlyHanseniasporaspp. in the later stages of oil palm wine tapping, vice versa in the case of ron palm wine tapping, with a unique presence ofSaccharomycodesin the later stages (15–30 days). Fructophilic lactic acid bacteria (FLAB), mainlyFructobacillusandLeuconostoc, encountered in both types of palm wine tapping showed a decline at later stages of oil palm wine tapping. In this type of palm wine, acetic acid bacteria with generaAcetobacterandGlucanoacetobacter, by surpassingLactobacillusin the last stage become dominant, whereasLactobacillusremained dominant in ron palm wine throughout tapping days. The decline in the relative abundance of gevotroline and essential amino acids during the later stages of palm wine tapping (15–25 days) supports the difference in the health benefits of the palm wine obtained from different days of tapping, indicating that early stages of tapping is more nutritional and healthy than the later stages. The microbial dynamics may be a potential indicator of metabolite changes during palm sap fermentation, thus contributing to establish particular features of palm wines in different stages of tapping. This understanding of microbial ecology and chemical composition changes during palm wine tapping can be used as biomarkers to assess palm wine’s quality and help to design an optimum starter culture.

The Cedars ultramafic block hosts alkaline springs (pH > 11) in which calcium carbonate forms upon uptake of atmospheric CO2 and at times via mixing with surface water. These processes lead to distinct carbonate morphologies with “floes” forming at the atmosphere-water interface, “snow” of fine particles accumulating at the bottom of pools and terraced constructions of travertine. Floe material is mainly composed of aragonite needles despite CaCO3 precipitation occurring in waters with low Mg/Ca ( The calcium carbonates exhibit an extreme range and approximately 1:1 covariation in δ13C (−9 to −28‰ VPDB) and δ18O (0 to −20‰ VPDB) that is characteristic of travertine formed in high pH waters. The large isotopic fractionations have previously been attributed to kinetic isotope effects accompanying CO2 hydroxylation but the controls on the δ13C-δ18O endmembers and slope have not been fully resolved, limiting the use of travertine as a paleoenvironmental archive. The limited areal extent of the springs (∼0.5 km2) and the limited range of water sources and temperatures, combined with our sampling strategy, allow us to place tight constraints on the processes involved in generating the systematic C and O isotope variations. We develop an isotopic reaction–diffusion model and an isotopic box model for a CO2-fed solution that tracks the isotopic composition of each dissolved inorganic carbon (DIC) species and CaCO3. The box model includes four sources or sinks of DIC (atmospheric CO2, high pH spring water, fresh creek water, and CaCO3 precipitation). Model parameters are informed by new floe Δ44Ca data (−0.75 ± 0.07‰), direct mineral growth rate measurements (4.8 to 8 × 10−7 mol/m2/s) and by previously published elemental and isotopic data of local water and DIC sources. Model results suggest two processes control the extremes of the array: (1) the isotopically light end member is controlled by the isotopic composition of atmospheric CO2 and the kinetic isotope fractionation factor (KFF (‰) = (α − 1) × 1000) accompanying CO2 hydroxylation, estimated here to be −17.1 ± 0.8‰ (vs. CO2(aq)) for carbon and −7.1 ± 1.1‰ (vs. ‘CO2(aq) + H2O’) for oxygen at 17.4 ± 1.0 °C. Combining our results with revised CO2 hydroxylation KFF values based on previous work suggests consistent KFF values of −17.0 ± 0.3‰ (vs. CO2(aq)) for carbon and −6.8 ± 0.8‰ for oxygen (vs. ‘CO2(aq) + H2O’) over the 17–28 °C temperature range. (2) The isotopically heavy endmember of calcium carbonates at The Cedars reflects the composition of isotopically equilibrated DIC from creek or surface water (mostly HC O 3 - , pH = 7.8–8.7) that occasionally mixes with the high-pH spring water. The bulk carbonate δ13C and δ18O values of modern and ancient travertines therefore reflect the proportion of calcium carbonate formed by processes (1) and (2), with process (2) dominating the carbonate precipitation budget at The Cedars. These results show that recent advances in understanding kinetic isotope effects allow us to model complicated but common natural processes, and suggest ancient travertine may be used to retrieve past meteoric water δ18O and atmospheric δ13C values. There is evidence that older travertine at The Cedars recorded atmospheric δ13C that predates large-scale combustion of fossil fuels.

The present study focuses on the rheological properties of polyethylene glycol (PEG) modified, positively charged, and negatively charged superparamagnetic iron oxide nanoparticles (SPIONs) at different temperatures. We hypothesized that the surface properties of these nanoparticles in the water did not affect their rheological properties. These nanoparticles had not the same surface properties as SPIONs-PEG had not to charge on their surface whereas positively charged and negatively charged ones with amine and carboxyl groups as their surfaces had positive and negative surface charges, respectively. However, their rheological behaviors were not different from each other. The comparative rheological study of SPIONs revealed their pseudo-Newtonian behavior. The viscosity of SPIONs decreased with the increase in temperature. At low shear rates, the shear stress of SPIONs was independent of rate and increased with the increase of rate. Moreover, at high shear rates, the shear stress for PEG-SPIONs was more than those for positively charged and negatively charged SPIONs. These measurements also revealed that at high shear rates, the shear stress of samples decreased with the increase of temperature. The shear stress of samples decreased with the increase of shear strain and the temperature. We also observed that all the samples had the same amount of shear strain at each shear stress, which indicated the exact resistance of SPIONs to deformation. Furthermore, the shear modulus decreased with time for these nanoparticles. These results suggest that these nanoparticles are promising candidates with appropriate properties for fluid processing applications and drug vectors in biomedical applications.

Background Reliable data on the adult SARS-CoV-2 infection fatality rate in Germany are still scarce. We performed a federal state-wide cross-sectional seroprevalence study named SaarCoPS, that is representative for the adult population including elderly individuals and nursing home residents in the Saarland. Methods Serum was collected from 2940 adults via stationary or mobile teams during the 1st pandemic wave steady state period. We selected an antibody test system with maximal specificity, also excluding seroreversion effects due to a high longitudinal test performance. For the calculations of infection and fatality rates, we accounted for the delays of seroconversion and death after infection. Results Using a highly specific total antibody test detecting anti-SARS-CoV-2 responses over more than 180 days, we estimate an adult infection rate of 1.02% (95% CI: [0.64; 1.44]), an underreporting rate of 2.68-fold (95% CI: [1.68; 3.79]) and infection fatality rates of 2.09% (95% CI: (1.48; 3.32]) or 0.36% (95% CI: [0.25; 0.59]) in all adults including elderly individuals, or adults younger than 70 years, respectively. Conclusion The study highlights the importance of study design and test performance for seroprevalence studies, particularly when seroprevalences are low. Our results provide a valuable baseline for evaluation of future pandemic dynamics and impact of public health measures on virus spread and human health in comparison to neighbouring countries such as Luxembourg or France.

Additional file 8: Figure S5. Multi-template matching is robust to noise. (A) Original image (2048 �� 2048 pixels). (B) Image as in A corrupted with artificial noise (normally distributed random noise ��� mean:0, standard deviation:50). (C, D) Result of multi-template matching for respectively A and B. The template used is a crop of the specimen in the middle of image A (hence a correlation score of 1 for the first row of Table C). Parameters for the detection: rotation of the template: 90,180�� - score type: 0-mean normalised cross-correlation - N = 4 expected objects per image ��� score threshold: 0.3 ��� maximal overlap between bounding boxes: 0.3.

Additional file 12: Figure S9. Using multi-template matching for phenotypes classification. (A) Manually annotated templates used for the classification of phenotypes of embryonic zebrafish kidneys. (B) Example of image to classify and (C) resulting correlation-scores for the 3 classes. In this case, the image is correctly classified as cystic. (D) Confusion matrix depicting the results for the classification of 167 annotated images (50 normal, 52 cystic, 65 long). The class Cystic and Normal are particularly well predicted. A number of Long were classified as Cystic, this can be expected as those 2 morphologies show similar elongated regions.

Additional file 9: Figure S6. Multi-template matching for eye-region detection in oriented zebrafish larvae. (A) Templates (108 �� 76 pixels) and image in which the search is performed (2048 �� 2048 pixels, scale bar: 1 mm). Orange rectangle indicates optional search region (1820 �� 452 pixels), and blue dotted rectangle the head-region template used for 2-step template matching (see B and D). Parameters for the detection: Vertical flipping of the templates (only if FlipV indicated), score type: 0-mean normalized cross-correlation, N = 2 expected objects per image, score threshold: 0.5, maximal overlap between bounding boxes: 0.25. For 2-step template matching the search region in orange is used for the 1st step (head-detection with a single head template, N = 1 expected object), then a single eye template is used with flipping for the detection of the eyes within the previously detected head region. (B) Result of the detections for N = 94 images. 2 eyes/1 eye/No eyes in figure legend refer to the outcome of eye-region detection in each larva. Vertical flipping of the templates readily increases the number of genuine matches. The 2-step template matching approach (search of template within a previously identified ROI) offers the best results and is recommended for more challenging template images (see Additional file 3). (C) Montage of the eye regions detections (yellow) for the 2-step matching approach as in B and D. Specimen in well B8 and F7 are excluded from the count in B as they are not dorsally oriented. (D) Mean computation time per image (error bars show standard deviation) for the different conditions (as in B) using the same hardware as in the main text.

Additional file 7: Figure S4. Template matching for head region detection in oriented zebrafish larvae. (A) Single template (188 �� 194 pixels, no additional transformation) and image (2048 �� 2048 pixels, scale bar: 1 mm) in which the search is performed. The orange rectangle shows the optionally used restricted search region (1820 �� 452 pixels). Parameters for the detection: score type: 0-mean normalised cross-correlation ��� N = 1 expected object per image. (B) Result of the detection for N = 96 images, with and without search region (both 100% detection rate). (C) Montage of detected zebrafish larval head regions within a 96 well plate (as in Fig. 1c). (D) Mean computation time per image (error bars show standard deviation) for the different conditions as in B using the same computing hardware as in the main text. Prior information about the position of the sample within the field of view (e.g. due to standardized sample mounting) can be used to specify a search region, drastically accelerating the computation and reducing the chance of incorrect predictions.

Additional file 5: Figure S2. Implementation in Fiji. (A) Graphical user interface for the plugin ���Template Matching Image��� with: (1) Dropdown menu to select the template image of the object of interest. The template must be smaller than the image specified in 2, (2) dropdown menu to select an image (or stack of images) in which to search for the template, (3) tick-boxes to optionally generate additional templates by horizontal/vertical flipping of the initial template, (4) input field for rotation angles to generate additional templates by rotations of the initial and, if selected, flipped templates. The angles are specified in degrees with clockwise orientation and must be separated by commas, (5) dropdown menu to choose the score used for the computation of the score map (normalised square-difference, normalised cross-correlation or 0-mean normalised cross-correlation), (6) input field to specify the number of objects expected in the image, (7) input field to enter a score-threshold in the range 0���1. If the normalised square-difference is selected, only local minima with values below the threshold are returned. While for cross-correlation scores, maxima above this value are returned, (8) input field to specify the maximum value in range 0���1 for the intersection over union (IoU) between a pair of overlapping bounding boxes (Non-Maxima Suppression), (9) tick-box to select if the detected Regions Of Interest (ROI) should be added to Fiji ROI Manager, (10) tick-box to specify if the result table should be displayed at the end of the execution. Parameters 7 and 8 are only required if several objects are expected in each image. (B) Outputs of the plugin with (1) result table with each row containing the names of the image and template, the prediction score and coordinates of the top left corner and centre of the predicted bounding box, and (2) the detected ROI appended to the ROI Manager and highlighted on the image (yellow).

Additional file 4: Figure S1. Flowchart of the implemented multi-template matching. The chart illustrates the sequential execution of the tool, for correlation-based score. For difference-based score, the pipeline is identical except that a difference map is computed, minima are detected instead of maxima and the lowest minima are returned. (IoU: Intersection over Union).x

Background: Head and neck cancer is particular due to its infiltrative nature and lymphatic extension, with multidisciplinary treatment. Immunotherapy may be a brand-new therapeutic approach.Case P...

Reliable data on the adult SARS-CoV-2 infection fatality rate in Germany are still scarce. We performed a federal state-wide cross-sectional seroprevalence study named SaarCoPS, that is representative for the adult population including elderly individuals and nursing home residents in the Saarland.Serum was collected from 2940 adults via stationary or mobile teams during the 1Using a highly specific total antibody test detecting anti-SARS-CoV-2 responses over more than 180 days, we estimate an adult infection rate of 1.02% (95% CI: [0.64; 1.44]), an underreporting rate of 2.68-fold (95% CI: [1.68; 3.79]) and infection fatality rates of 2.09% (95% CI: (1.48; 3.32]) or 0.36% (95% CI: [0.25; 0.59]) in all adults including elderly individuals, or adults younger than 70 years, respectively.The study highlights the importance of study design and test performance for seroprevalence studies, particularly when seroprevalences are low. Our results provide a valuable baseline for evaluation of future pandemic dynamics and impact of public health measures on virus spread and human health in comparison to neighbouring countries such as Luxembourg or France.

The Cedars ultramafic block hosts alkaline springs (pH > 11) in which calcium carbonate forms upon uptake of atmospheric CO2 and at times via mixing with surface water. These processes lead to distinct carbonate morphologies with “floes” forming at the atmosphere-water interface, “snow” of fine particles accumulating at the bottom of pools and terraced constructions of travertine. Floe material is mainly composed of aragonite needles despite CaCO3 precipitation occurring in waters with low Mg/Ca (

Quantitative measurements and qualitative description of scientific images are both important to describe the complexity of digital image data. While various software solutions for quantitative measurements in images exist, there is a lack of simple tools for the qualitative description of images in common user-oriented image analysis software. To address this issue, we developed a set of Fiji plugins that facilitate the systematic manual annotation of images or image-regions. From a list of user-defined keywords, these plugins generate an easy-to-use graphical interface with buttons or checkboxes for the assignment of single or multiple pre-defined categories to full images or individual regions of interest. In addition to qualitative annotations, any quantitative measurement from the standard Fiji options can also be automatically reported. Besides the interactive user interface, keyboard shortcuts are available to speed-up the annotation process for larger datasets. The annotations are reported in a Fiji result table that can be exported as a pre-formatted csv file, for further analysis with common spreadsheet software or custom automated pipelines. To illustrate possible use case of the annotations, and facilitate the analysis of the generated annotations, we provide examples of such pipelines, including data-visualization solutions in Fiji and KNIME, as well as a complete workflow for training and application of a deep learning model for image classification in KNIME. Ultimately, the plugins enable standardized routine sample evaluation, classification, or ground-truth category annotation of any digital image data compatible with Fiji.

Despite widespread drug exposure, for example during gestation or in prematurely born children, organ-specific developmental toxicity of most drugs is poorly understood. Developmental and functional abnormalities are a major cause of kidney diseases during childhood; however, the potential causal relationship to exposure with nephrotoxic drugs during nephrogenesis is widely unknown. To identify developmental nephrotoxic drugs in a large scale, we established and performed an automated high-content screen to score for phenotypic renal alterations in theTg(wt1b:EGFP)zebrafish line. During early nephrogenesis, embryos were exposed to a compound library of approved drugs. After treatment, embryos were aligned within microtiter plates using 3D-printed orientation tools enabling the robust acquisition of consistent dorsal views of pronephric kidneys by automated microscopy. To qualitatively and quantitatively score and visualize phenotypes, we developed software tools for the semi-automated analysis, processing and visualization of this large image-based dataset. Using this scoring scheme, we were able to categorize compounds based on their potential developmental nephrotoxic effects. About 10% of tested drugs induced pronephric phenotypes including glomerular and tubular malformations, or overall changes in kidney morphology. Major chemical compound groups identified to cause glomerular and tubular alterations included dihydropyridine derivatives, HMG CoA reductase inhibitors, fibrates, imidazole, benzimidazole and triazole derivatives, corticosteroids, glucocorticoids, acetic acid derivatives and propionic acid derivatives. In conclusion, the presented study demonstrates the large-scale screening of kidney-specific toxicity of approved drugs in a live vertebrate embryo. The associated technology and tool-sets can be easily adapted for other organ systems providing a unique platform forin vivolarge-scale assessment of organ-specific developmental toxicity or other biomedical applications. Ultimately, the presented data and associated visualization and browsing tools provide a resource for potentially nephrotoxic drugs and for further investigations.

The kepler and TESS missions have generated over 100,000 potential transit signals that must be processed in order to create a catalog of planet candidates. During the last few years, there has been a growing interest in using machine learning to analyze these data in search of new exoplanets. Different from the existing machine learning works, ExoMiner, the proposed deep learning classifier in this work, mimics how domain experts examine diagnostic tests to vet a transit signal. ExoMiner is a highly accurate, explainable, and robust classifier that 1) allows us to validate 301 new exoplanets from the MAST Kepler Archive and 2) is general enough to be applied across missions such as the on-going TESS mission. We perform an extensive experimental study to verify that ExoMiner is more reliable and accurate than the existing transit signal classifiers in terms of different classification and ranking metrics. For example, for a fixed precision value of 99%, ExoMiner retrieves 93.6% of all exoplanets in the test set (i.e., recall=0.936) while this rate is 76.3% for the best existing classifier. Furthermore, the modular design of ExoMiner favors its explainability. We introduce a simple explainability framework that provides experts with feedback on why ExoMiner classifies a transit signal into a specific class label (e.g., planet candidate or not planet candidate)., Comment: Accepted for Publication in Astrophysical Journals, November 12, 2021

The δ18O of ostracod valves is widely used to infer water δ18O and temperature. However, ostracod δ18O appears sensitive to other environmental variables. In addition, there is species-dependent ostracod calcite 18O enrichment, relative to slowly precipitated inorganic calcite under the same conditions. Together these uncertainties complicate ostracod paleoclimate reconstructions. This study presents a new understanding of the causes of ostracod δ18O variations based on a global database of published ostracod δ18O values in lake, marine and coastal environments, and from culture experiments. The database includes associated field/experiment host water parameters including temperature (−1 to 32 °C), water δ18O (−13.2‰ to 4.3‰ VSMOW), pH (6.9–10.4), salinity (0–72 g/kg), calcite saturation states (0.6–26), and dissolved inorganic carbon concentration [DIC] (0.9–54.3 mmol/kg). The data show that: (1) the δ18O of marine and non-marine ostracods reflects the 18O/16O of the sum of host water CO32− and HCO3− ions. For example, at a given temperature, the δ18O of non-marine ostracods decreases by 4‰ to 6‰ as [CO32−]/[DIC] reaches 70%, depending on the ostracod species. In low [CO32−]/[DIC] settings (i.e. high HCO3−/CO32−), ostracod 18O/16O is close to the 18O/16O of HCO3− ions, which explains why on average ostracod δ18O is higher than the δ18O of inorganic calcite precipitated slowly under the same conditions. (2) Taxonomic offsets in ostracod δ18O vary with the host water [CO32−]/[DIC]. In environments where HCO3− ≫ CO32− (i.e. most freshwater lakes), the 18O/16O of Candonids is indistinguishable from the 18O/16O of HCO3− ions (difference of 0.10 ± 0.16‰) while the 18O/16O of Cyprids is lower than the 18O/16O of HCO3− ions by −0.77‰ to −0.32‰, Cytherids by −0.88 ± 0.29‰, and Limnocytherids by −1.12 ± 0.05‰. (3) The sensitivity of ostracod δ18O to [CO32−]/[DIC] also varies with taxonomy. For each percent increase in [CO32−]/[DIC], Candonids δ18O decreases by −0.098 ± 0.024‰, Cyprids by −0.075 ± 0.004‰, Cytherids by −0.057 ± 0.012‰, and Limnocytherids by −0.058 ± 0.005‰. (4) The similarity in 18O/16O between ostracod calcite and the sum of host water ‘HCO3− and CO32−’ (despite species-specific offsets) suggests rapid precipitation of a finite DIC pool in the ostracod calcifying fluid. We propose an ostracod δ18O model whereby the more negative the 18O/16O of a given species relative to the sum of CO32− and HCO3−, the greater the isotopic equilibrium between the DIC pool and H2O in the ostracod calcifying fluid. (5) Higher host water salinities and pH induce higher [CO32−]/[DIC], resulting in lower ostracod δ18O, which explains the variable salinity and pH effects on ostracod δ18O. Our database and ostracod δ18O model shows that ostracod δ18O records from closed basin environments likely reflect high or variable [CO32−]/[DIC], rather than water temperature and δ18O alone. Our study reconciles contradictory observations of controls on ostracod δ18O and paves the way for improved paleo-environmental interpretations and reconstructions of past water [CO32−]/[DIC].

Trabectedin (ET-743) is a marine alkaloid isolated from the Caribbean tunicate Ecteinascidia turbinata, with a chemical structure characterized by three fused tetrahydroisoquinoline rings. In the present case report, two patients with advanced and metastatic uterine leiomyosarcomas (ULMS) with significant progression-free survival (PFS) and overall survival (OS) administered Trabectedin as second and third line treatment are reported. The first case received third line Trabectedin with a PFS of 24 months and an OS of 35 months. The second case received second line Trabectedin with a PFS of 24 months and an OS of 30 months. In addition, a good safety record was obtained in the long-term administration of Trabectedin (more so in case 1 than case 2), with a good quality of life.

Following recent experiments on ultracold dual superflows, we model in this work the dynamics of two harmonically trapped counterflowing superfluids. Using complementary analytical and numerical approaches, we study the shedding of elementary excitations triggered by the relative motion of the two species. We exhibit two different excitation mechanisms leading to distinct threshold velocities for the onset of dissipation: in addition to the parametric pair production present in homogeneous , galilean-invariant systems, we show that non-uniform motion and density inhomogeneities allow for a Landau-like decay mechanism where single excitations are produced.

Despite concerns over toxicity, carbon nanotubes have been extensively investigated for potential applications in nanomedicine because of their small size, unique properties, and ability to carry cargo such as small molecules and nucleic acids. Herein, we show that polymer nanotubes can be synthesized quickly and easily from a homopolymer of ethylene glycol dimethacrylate (EGDMA). The nanotubes formed via photo‐initiated polymerization of the highly functional prepolymer, inside an anodized aluminium oxide template, have a regular structure and large internal pore and can be loaded with a fluorescent dye within minutes representing a simple alternative to multi‐walled carbon nanotubes for biomedical applications.

Metastasis of non-gynaecological tumours to the cervix is a rare event, and metastasis from breast cancer is even rarer, with only a limited number of such cases reported in the literature to date. We herein report the case of an 86-year-old female patient who had undergone mastectomy and axillary lymphadenectomy for invasive ductal cell breast carcinoma 2 years prior, followed by adjuvant hormonal therapy with letrozole. During hospitalization for anemia associated with an inflammatory syndrome and abdominal pain with menorrhagia, an abdominal ultrasound examination revealed a suspicious uterine mass with irregular contours and abnormal vascularization with associated increase of the blood level of cancer antigen 15-3 to 34 kU/l. The histological and immunohistochemical analysis of a cervical biopsy sample discover a secondary lesion metastatic from the primary ductal cell breast carcinoma. The metastatic tissue was hormone-negative, which was compatible with disease progression during hormonal therapy. Considering the multiple metastasis, comorbidities, unfavourable performance status and the quick deterioration of the patient's clinical condition, only best supportive care was administered.

Summary Listeria monocytogenes is a bacterial pathogen classified into distinct serovars (SVs) based on somatic and flagellar antigens. To correlate phenotype with genetic variation, we analyzed the wall teichoic acid (WTA) glycosylation genes of SV 1/2, 3 and 7 strains, which differ in decoration of the ribitol-phosphate backbone with N-acetylglucosamine (GlcNAc) and/or rhamnose. Inactivation of lmo1080 or the dTDP-l-rhamnose biosynthesis genes rmlACBD (lmo1081–1084) resulted in loss of rhamnose, whereas disruption of lmo1079 led to GlcNAc deficiency. We found that all SV 3 and 7 strains actually originate from a SV 1/2 background, as a result of small mutations in WTA rhamnosylation and/or GlcNAcylation genes. Genetic complementation of different SV 3 and 7 isolates using intact alleles fully restored a characteristic SV 1/2 WTA carbohydrate pattern, including antisera reactions and phage adsorption. Intriguingly, phage-resistant L. monocytogenes EGDe (SV 1/2a) isolates featured the same glycosylation gene mutations and were serotyped as SV 3 or 7 respectively. Again, genetic complementation restored both carbohydrate antigens and phage susceptibility. Taken together, our data demonstrate that L. monocytogenes SV 3 and 7 originate from point mutations in glycosylation genes, and we show that phage predation represents a major driving force for serovar diversification and evolution of L. monocytogenes.

Specific clusters of metabolic syndrome (MetS) components impact differentially on arterial stiffness, indexed as pulse wave velocity (PWV). Of note, in several population-based studies participating in the MARE (Metabolic syndrome and Arteries REsearch) Consortium the occurrence of specific clusters of MetS differed markedly across Europe and the US. The aim of the present study was to investigate whether specific clusters of MetS are consistently associated with stiffer arteries in different populations. We studied 20,570 subjects from 9 cohorts representing 8 different European countries and the US participating in the MARE Consortium. MetS was defined in accordance with NCEP ATPIII criteria as the simultaneous alteration in >= 3 of the 5 components: abdominal obesity (W), high triglycerides (T), low HDL cholesterol (H), elevated blood pressure (B), and elevated fasting glucose (G). PWV measured in each cohort was "normalized" to account for different acquisition methods. MetS had an overall prevalence of 24.2% (4985 subjects). MetS accelerated the age-associated increase in PWV levels at any age, and similarly in men and women. MetS clusters TBW, GBW, and GTBW are consistently associated with significantly stiffer arteries to an extent similar or greater than observed in subjects with alteration in all the five MetS components - even after controlling for age, sex, smoking, cholesterol levels, and diabetes mellitus - in all the MARE cohorts. In conclusion, different component clusters of MetS showed varying associations with arterial stiffness (PWV). (C) 2014 Elsevier Ireland Ltd. All rights reserved., NIH -National Institutes of Health

Journal based metrics is known not to be ideal for the measurement of the quality of individual researcher's scientific output. In the current report 16 contributors from Hong Kong SAR, India, Korea, Taiwan, Russia, Germany, Japan, Turkey, Belgium, France, Italy, UK, The Netherlands, Malaysia, and USA are invited. The following six questions were asked: (I) is Web of Sciences journal impact factor (IF) and Institute for Scientific Information (ISI) citation the main academic output performance evaluation tool in your institution? and your country? (II) How does Google citation count in your institution? and your country? (III) If paper is published in a non-SCI journal but it is included in PubMed and searchable by Google scholar, how it is valued when compared with a paper published in a journal with an IF? (IV) Do you value to publish a piece of your work in a non-SCI journal as much as a paper published in a journal with an IF? (V) What is your personal view on the metric measurement of scientific output? (VI) Overall, do you think Web of Sciences journal IF is beneficial, or actually it is doing more harm? The results show that IF and ISI citation is heavily affecting the academic life in most of the institutions. Google citation and evaluation, while is being used and convenient and speedy, has not gain wide 'official' recognition as a tool for scientific output evaluation.

The addition of ezetimibe, an intestinal cholesterol absorption inhibitor, to statin therapy has recently shown clinical benefits in the Improved Reduction of Outcomes: Vytorin Efficacy International Trial by reducing low-density-lipoprotein (LDL) cholesterol levels more than statin therapy alone. Here, we investigated the mechanisms by which inhibition of intestinal cholesterol absorption might contribute to the clinically observed reduction in cardiovascular events by evaluating its effect on inflammatory plaque development in apolipoprotein E

Background Since patients with diabetes mellitus have increased rates of cardiovascular morbidity and mortality, it is critical to evaluate cardiovascular risk perioperatively. Although several preoperative risk indices have been developed, including the Goldman and Detsky indices, none have been designed specifically for diabetic patients. In this study, we attempted to identify predictors of postoperative cardiac and noncardiac complications in diabetic patients undergoing elective general surgery. Study Design A cohort of 107 diabetic patients undergoing elective surgery was assembled. Basic demographic and clinical data were recorded perioperatively, and all patients were followed up prospectively daily for 7 days postoperatively. Patients were interviewed at 5 years postoperatively. Univariate and multivariate analyses were performed to identify predictors of postoperative complications and long-term cardiac and vascular morbidity and mortality. Results Total cardiac complications and myocardial infarction were predicted by the Goldman index. Wound complications were not predicted by any of the variables studied. At 5 years postoperatively, cardiac and vascular deaths and events were predicted by age, history of myocardial infarction or stroke, presence of vascular disease, Goldman index, duration of diabetes or hypertension, Charlson comorbidity score, and postoperative myocardial infarction or cardiac arrest. Blood sugar control during surgery was not predictive of any short- or long-term cardiovascular complications. Conclusions Total cardiac complications had a significant preoperative predictor: the Goldman index. Both preoperative and postoperative variables and indices predicted long-term cardiac and vascular complications. Further study is necessary to investigate these relationships to better assess and manage the diabetic patient in the perioperative setting.

PubMed ID: 25112663, Aims: Estimated central systolic blood pressure (cSBP) and amplification (Brachial SBP-cSBP) are non-invasive measures potentially prognostic of cardiovascular (CV) disease. No worldwide, multiple-device reference values are available. We aimed to establish reference values for a worldwide general population standardizing between the different available methods of measurement. How these values were significantly altered by cardiovascular risk factors (CVRFs) was then investigated. Methods and results: Existing data from population surveys and clinical trials were combined, whether published or not. Reference values of cSBP and amplification were calculatedas percentiles for 'Normal' (no CVRFs) and 'Reference' (any CVRFs) populations. We included 45 436 subjects out of 82 930 that were gathered from 77 studies of 53 centres. Included subjects were apparently healthy, not treated for hypertension or dyslipidaemia, and free from overt CV disease and diabetes. Values of cSBP and amplification were stratified by brachial blood pressure categories and age decade in turn, both being stratifiedbysex. Amplification decreased with age and more so in males than in females. Sex was the most powerful factor associated with amplification with 6.6 mmHg (5.8-7.4) higher amplification in males than in females. Amplification was marginally but significantly influenced by CVRFs, with smoking and dyslipidaemia decreasing amplification, but increased with increasing levels of blood glucose. Conclusion: Typical values of cSBP and amplification in a healthy population and a population free of traditional CVRFs are now available according to age, sex, and brachial BP, providing values included from different devices with a wide geographical representation. Amplification is significantly influenced by CVRFs, but differently in men and women. © 2014 Published on behalf of the European Society of Cardiology.

To more clearly define the scope and impact of violence in health care facilities, national data on assaults in VA medical centers and freestanding clinics were examined.A survey was distributed to all VA medical centers and freestanding clinics asking for cumulative data for one fiscal year (October 1990 through September 1991). Data were obtained on number, types, and locations of physical assaults and other assaultive behavior; the types of staff assaulted and number of workdays lost due to injuries; diagnoses of perpetrators; recommendations made after the incidents were reviewed; training in prevention and management of assaultive behavior; and the impact of training on rates of assaultive behavior.During the survey year, 24,219 incidents of assaultive behavior were reported by 166 VA facilities; 8,552 incidents involved battery or physical assault. Weapon possession by perpetrators was common (8.5 percent of incidents), and weapons were used in 130 assaults (1.5 percent of assaults). Assaults occurred most frequently in psychiatric units (43.1 percent), followed by long-term-care units (18.5 percent) and admitting or triage areas (13.4 percent). Assault-related injuries were most common among nursing personnel. Perpetrators of assaults were most typically diagnosed as having psychoses, substance use disorders, or dementia. On inpatient psychiatry units, an inverse correlation was found between expenditures on staffing and the frequency of assaultive incidents. Staff training on management of assaultive behavior varied widely.Assaultive behavior is a significant problem for health care workers. Staff in all clinical areas need to be prepared to deal with assaultive patients. More research is needed on staff training and interventions for preventing and limiting assaults.

The study aim was to determine the prevalence of repeated assaults on staff and other patients and characteristics of patients who commit repeated assaults in the Veterans Health Administration of the Department of Veterans Affairs.Patients in VA medical centers and freestanding outpatient clinics who committed two or more assaults in fiscal years 1995 and 1996 were identified through a survey of facility quality or risk managers. For each repeatedly assaultive patient, structured information, including incident reports, was obtained for all assault occasions.A total of 153 VA facilities responded, for a response rate of 99 percent. The survey identified 8,968 incidents of repeated assault by 2,233 patients, for a mean of 4.02 assaults per patient in the two-year study period. In 92 percent of the incidents, the assaultive patient had a primary or secondary psychiatric diagnosis. The mean age of the repeat assaulters was 62 years. Ninety-eight percent of the repeat assaulters were male, and 76.6 percent were Caucasian. At least 16 percent of the assaulters, 22 percent of the patients assaulted, and 20 percent of the staff assaulted required medical attention for injuries, which, along with the number of lost work days, indicates that repeated assaults are costly.Repeatedly assaultive patients represent major challenges to their own safety as well as to that of other patients and staff. Identifying patients at risk for repeated assaults and developing intervention strategies is critically important for ensuring the provision of health care to the vulnerable population of assaultive patients.

SummaryStaphylokinase (Sak), a 15.5-kDa bacterial protein, forms a complex with human plasmin, which in turn activates other plasminogen molecules to plasmin. Three recombinant DNA-based approaches, (i) site directed substitution with alanine, (ii) search for proximity relationships at the complex interface, and (iii) active-site accessibility to protease inhibitors have been used to deduce a coherent docking model of the crystal structure of Sak on the homology-based model of micro-plasmin (μPli), the serine protease domain of plasmin. Sak binding on μPli is primarily mediated by two surface-exposed loops, loops 174 and 215, at the rim of the active-site cleft, while the binding epitope of Sak on μPli involves several residues located in the flexible NH2-terminal arm and in the five-stranded mixed β-sheet. Several Sak residues located within the unique μ-helix and the β2 strand do not contribute to the binding epitope but are essential to induce plasminogen activating potential in the Sak:μPli complex. These residues form a topologically distinct activation epitope, which, upon binding of Sak to the catalytic domain of μPli, protrudes into a broad groove near the catalytic triad of μPli, thereby generating a competent binding pocket for micro-plasminogen (μPlg), which buries approximately 2500 Å of the Sak:μPli complex upon binding. This structural and functional model may serve as a template for the design of improved Sak-derived thrombolytic agents. Following the completion and presentation of the present study, the deduced Sak:μPli:μPlg complex was fully confirmed by X-ray crystallography, which further illustrates the power and potential of the present approach.

Staphylokinase (Sak), a 16-kDa bacterial protein, forms a 1:1 stoichiometric complex with the serine proteinase domain of human plasmin, which in turn converts other plasminogen molecules into plasmin. To identify amino acid residues critical for generating the Sak:plasmin activator complex, alanine-scanning mutagenesis was performed on phage-displayed micro-plasminogen (microPlg). Substitution of Arg719 with Ala [microPlg(R719A)] disrupted complex formation, although the sensitivity of phage-displayed microPlg(R719A) to activation by urokinase and the amidolytic activity of the micro-plasmin derivative [microPli(R719A)] remained unaffected. Likewise, the soluble microPlg(R719A) molecule did not generate a functional activator complex with Sak, whereas quantitative activation into plasmin was obtained upon incubation with either urokinase or the Sak:plasmin complex. Real-time biospecific affinity measurements revealed that the Arg --> Ala substitution at position 719 increased the equilibrium dissociation constant between microPlg(R719A) and Sak from 46 nM to 1 microM, primarily by reducing the association rate constant. Arg719 has recently also been implied in the functional complex formation between human plasmin and streptokinase [Dawson, K. M., Marshall, J. M., Raper, R. H., Gilbert, R. J., and Ponting, C. P. (1994) Biochemistry 33, 12042-12047.], suggesting that both bacterial cofactors may share common structural and/or mechanistic aspects for plasminogen activation.

The effectiveness of clozapine treatment in a treatment-refractory sample of older adult veterans with primary psychosis was examined.Data were collected over a five-year period for patients age 55 and older who were given clozapine because of a history of treatment-refractory or treatment-intolerant psychosis. At initiation of clozapine therapy, baseline demographic, clinical, and psychopathology data were collected. At baseline and quarterly, patients' psychopathology was rated with the Brief Psychiatric Rating Scale (BPRS), and involuntary movements were rated with the Abnormal Involuntary Movement Scale (AIMS).The 329 patients age 55 or older who received clozapine during the study period represented 10 percent of all patients on clozapine therapy in the VA system. Of the 312 patients for whom demographic information was available, 294 were men and 18 were women. Overall, patients improved on clozapine therapy, although wide variation in drug response was observed. Complete BPRS and AIMS data were available for 97 patients. The 55- to 64-year-old group had a mean improvement in total BPRS score of 19.8 percent, with 42.6 percent showing more than a 20 percent improvement; those age 65 and older had a mean improvement of 5.7 percent, with 17.2 percent showing an improvement greater than 20 percent. The 97 patients with complete AIMS data showed a mean improvement of 16.6 percent in total score.Clozapine is an important therapeutic agent for older adults with treatment-refractory psychosis. Patients between the ages of 55 and 64 may have a better response than those age 65 and older.

Since most antibodies directed against protein antigens recognize epitopes composed of several discontinuous segments of the polypeptide chain, attempts to delineate the amino acids constituting these epitopes with the use of linear peptides have generally been unsuccessful. Here, a method is described based on error-prone PCR, phage display and negative selection, whereby amino acid residues constituting the functional epitope are identified in the context of the native protein. First a library of randomized antigen variants containing most single, double and triple amino acid mutants generated by single nucleotide substitutions is produced by error-prone PCR amplification of the DNA sequence encoding the protein antigen. The phage-displayed library is then negatively selected for epitope loss mutants by passing through an affinity matrix derivatized with a specific antibody and positively selected for retention of function. This method was applied to the mapping of the epitopes of two murine monoclonal antibodies (MA-7H11 and MA-3G10) on staphylokinase, a 136 amino acid plasminogen activator secreted by some strains of Staphylococcus aureus. After two negative/positive selection cycles, DNA sequencing of several clones revealed preferential amino acid mutations at positions 35 and 130 (with MA-7H11), and at positions 62, 66 and 136 (with MA-3G10). Affinity measurements of staphylokinase variants carrying single amino acid mutations at these positions confirmed their contribution to the free energy of binding to MA-7H11 and MA-3G10. This approach may be useful for isolating mutants with altered antigenic or functional properties and in general to map critical regions for protein-protein interactions.

Background The “charged cluster–to-alanine” substitution variants SakSTAR(K35A,E38A,K74A,E75A,R77A) and SakSTAR(K74A,E75A,R77A,E80A,D82A), previously identified as SakSTAR.M38 and SakSTAR.M89, respectively, induce less antibody formation in patients than wild-type recombinant staphylokinase (SakSTAR), but their specific activities are reduced by 50%. Therefore, the effect of the reversal of one or more of these substituted amino acids on the ratio of activity to antigenicity was studied. Methods and Results Fourteen mutants with one to four “alanine-to–wild-type” reversals were expressed in Escherichia coli and highly purified (>95%). In rabbits immunized with wild-type SakSTAR, the combined K35,E38,K74,E75,R77 or K74,E75,R77,E80,E82 epitope accounted for only 30% of antibody absorption from plasma, and no clear immunodominant residue could be identified. In baboons immunized with SakSTAR, the K35,E38 and K74,E75,R77 epitopes or the K74,E75,R77 and E80,D82 epitopes contributed equally to account for 50% of total antibody binding, but no immunodominant residues were apparent. In pooled plasma from patients with peripheral arterial occlusion treated with wild-type SakSTAR, about 40% of the antibodies depended on K74 of epitope K74,E75,R77 for binding, whereas epitopes K35,E38 and E80,D82 had a negligible contribution toward antibody recognition. Conclusions The recognition pattern by SakSTAR variants of antibodies induced with wild-type SakSTAR differs markedly among species. This implies that a systematic evaluation of reduced antigen recognition and antibody induction in humans will require the development of human or humanized systems. Surprisingly, SakSTAR(K 74 ), with a single substitution of Lys74 with Ala, had an intact specific activity but did not absorb 40% of the antibodies induced in patients by treatment with wild-type SakSTAR.

This article describes an economically designed microclimate vitrine for panel paintings, which can be easily produced in-house with minimal aesthetic compromise to the paintings. The design uses the picture's frame as the primary housing in combination with museum glazing and a backplate to create a sealed microclimate vitrine. It is also adaptable to other framed artworks (canvas, paper, parchment) in need of microclimate protection. A summary is given of tests carried out at the Central Research Laboratory in Amsterdam using this design and another more costly commonly used design. The latter consisted of a closed box made of inert materials containing the painting, which in turn is set into the picture's frame. Testing showed that the two designs perform with similar efficiency. Further, the use of silica gel in a closed system with minimized air space is reevaluated. A design plan and materials are included.

To (1) synthesize the terminology used to classify extramedullary spinal cysts in dogs to clarify some of the commonly reported misconceptions, and (2) propose a classification scheme to limit confusion with terminology.Literature review.An online bibliographic search was performed in January 2013 for articles relating to extramedullary spinal cysts in dogs using PubMed (http://www.pubmed.gov/) and Google Scholar (http://scholar.google.com/) databases. Only peer-reviewed clinical literature describing cystic lesions pertaining to the spinal cord and associated structures was included.From 1962 to 2013, 42 articles were identified; 25 (95 dogs) reported meningeal cysts, 10 (24 dogs) described 60 extradural cysts, 3 reports (18 dogs) described discal cysts or acute compressive hydrated nucleus pulposus extrusions (HNPE). Spinal cysts were categorized by location based on cross-sectional imaging as meningeal or extradural non-meningeal. Sub-classification was then performed based on surgical findings and pathology. Meningeal cysts included arachnoid diverticulae and Tarlov (perineural) cysts. Extradural non-meningeal cysts included intraspinal cysts of the vertebral joints, ligaments and discs. Discal cysts also fit this category and have been reported extensively in humans but appear rare in dogs.Extramedullary spinal cysts should be first classified according to location with a sub-classification according to pathologic and surgical findings. Previous canine cases of discal cysts appear to represent a different disease entity and the term acute compressive HNPE is therefore preferred.

We describe a novel phage display system that affords the surface expression and hence affinity selection of cDNAs. The strategy is based on a new approach to functionally display proteins on filamentous phage through the attachment to the C-terminus of the minor coat protein VI. The utility of the method was evaluated using a cDNA library derived from the parasite Ancylostoma caninum. cDNA sequences were fused in each of the three reading frames to the 3'-end of the M13 gene VI expressed by a phagemid vector. Phages rescued from this cDNA expression library were subjected to biopanning against two serine proteases, trypsin and the human coagulation factor Xa. This led to the identification of cDNAs encoding novel members of two different families of serine protease inhibitors. The authenticity of the cDNA selected with trypsin as the target was demonstrated by purifying the encoded potent Kunitz-type inhibitor from an Ancylostoma caninum extract. The rapid isolation of specific cDNAs with the protein VI monovalent display system should facilitate the search for novel biologically important ligands.