bluuebunny/crossref_metadata_2025 huggingface dataset:
Relations of Ruminal Fermentation Parameters and Microbial Matters to Odd- and Branched-Chain Fatty Acids in Rumen Fluid of Dairy Cows at Different Milk Stages
Authors: Liu Keyuan , Li Yang , Luo Guobin , Xin Hangshu , Zhang Yonggen , Li Guangyu
Published: November 2019

The purpose of this research was to evaluate whether relationships exist between odd- and branched-chain fatty acids (OBCFAs) originating from milk fat and the corresponding data of ruminal fermentation parameters, microbial populations, and base contents that were used to mark microbial protein in rumen. Nine lactating Holstein dairy cows with similar body weights and parity were selected in this study, and the samples of rumen and milk were collected at the early, middle, and late stages, respectively. The rumen and milk samples were collected over three consecutive days from each cow, and the ruminal and milk OBCFA profiles, ruminal fermentation parameters, bacterial populations, and base contents were measured. The results showed that the concentrations of OBCFAs, with the exception of C11:0 and C15:0, were significantly different between milk and rumen (p < 0.05). The concentrations of anteiso-fatty acids in milk were higher than those in rumen, and the contents of linear odd-chain fatty acids were higher than those of branched-chain fatty acids in both milk and rumen. Significant relationships that existed between the concentrations of C11:0, iso-C15:0, anteiso-C15:0, C15:0, and anteiso-C17:0 in rumen and milk (p < 0.05). The total OBCFA content in milk was positively related to the acetate molar proportion but negatively correlated with isoacid contents (p < 0.05). The populations of Ruminococcus albus, R. flavefacients, and Eubacterium ruminantium were significantly related to milk C13:0 contents (p < 0.05). The adenine/N ratio was negatively related to milk OBCFA content (p < 0.05) but positively associated with the iso-C15:0/iso-C17:0 ratio (p < 0.05). Milk OBCFAs were significantly correlated with ruminal fermentation parameters, ruminal bacterial populations, and base contents. Milk OBCFAs had the potential to predict microbial nitrogen flow, and the prediction equations for ruminal microbial nitrogen flow were established for OBCFAs in dairy milk.
Dietary Stevioside Supplementation Alleviates Lipopolysaccharide-Induced Intestinal Mucosal Damage through Anti-Inflammatory and Antioxidant Effects in Broiler Chickens
Authors: Jiang Jingle , Qi Lina , Lv Zengpeng , Jin Song , Wei Xihui , Shi Fangxiong
Published: November 2019

The study was conducted to investigate the effects of dietary stevioside (STE) supplementation on the lipopolysaccharide (LPS)-induced intestinal mucosal damage of broiler chickens. A total of 192 one-day-old male Ross 308 broiler chicks were randomly divided into four treatments: (1) basal diet (CON); (2) basal diet supplemented with 250 mg/kg stevioside (STE); (3) basal diet + LPS-challenge (LPS); (4) basal diet supplemented with 250 mg/kg stevioside + LPS-challenge (LPS + STE). LPS-challenged groups received an intraperitoneal injection of LPS at 17, 19 and 21 d, whereas the CON and STE groups received a saline injection. The results showed that dietary STE supplementation normalized LPS-induced changes in protein expression of p-NF-κB and p-IκBα, mRNA expression of inflammatory genes (TLR4, NF-κB, and IFN-γ), tight junction-related genes (CLDN2, OCLN, and ZO-1), and antioxidant genes (Nrf2 and HO-1). LPS-induced decreases in serum diamine oxidase (DAO) level, villus height-to-crypt depth ratio, apoptotic index, and protein expression of proliferating cell nuclear antigen (PCNA) were reversed with dietary STE supplementation. Additionally, STE supplementation ameliorated the redox damage by reducing malondialdehyde (MDA) content and increasing total antioxidant capacity (T-AOC) and antioxidant enzyme activity. In conclusion, dietary stevioside supplementation could alleviate LPS-induced intestinal mucosal damage through anti-inflammatory and antioxidant effects in broiler chickens.
Experimental Investigation of Concrete with Recycled Aggregates for Suitability in Concrete Structures
Authors: Denisiewicz Arkadiusz , Śliwa Małgorzata , Kula Krzysztof , Socha Tomasz
Published: November 2019

This paper presents the experimental tests of concrete made on the recycled aggregates basis. Tests were carried out to determine the concrete suitability for construction purposes. The physical and strength properties were determined for three types of recycling aggregates. The aggregates were obtained from sanitary ceramics ‘SC’ (washbasins and toilet bowls), building ceramics ‘BC’ (solid bricks), and concrete rubble ‘CR’. The results obtained in tests of compressive strength, bending tensile strength, water absorption, total shrinkage, watertightness, and frost resistance of concrete made of SC and CR aggregates gave grounds for stating its suitability for structural purposes. Concrete based on the BC aggregates is not recommended for structural applications.
Green Production and Biotechnological Applications of Cell Wall Lytic Enzymes
Authors: Benedetti Manuel , Locci Federica , Gramegna Giovanna , Sestili Francesco , Savatin Daniel V.
Published: November 2019

Energy demand is constantly growing, and, nowadays, fossil fuels still play a dominant role in global energy production, despite their negative effects on air pollution and the emission of greenhouse gases, which are the main contributors to global warming. An alternative clean source of energy is represented by the lignocellulose fraction of plant cell walls, the most abundant carbon source on Earth. To obtain biofuels, lignocellulose must be efficiently converted into fermentable sugars. In this regard, the exploitation of cell wall lytic enzymes (CWLEs) produced by lignocellulolytic fungi and bacteria may be considered as an eco-friendly alternative. These organisms evolved to produce a variety of highly specific CWLEs, even if in low amounts. For an industrial use, both the identification of novel CWLEs and the optimization of sustainable CWLE-expressing biofactories are crucial. In this review, we focus on recently reported advances in the heterologous expression of CWLEs from microbial and plant expression systems as well as some of their industrial applications, including the production of biofuels from agricultural feedstock and of value-added compounds from waste materials. Moreover, since heterologous expression of CWLEs may be toxic to plant hosts, genetic strategies aimed in converting such a deleterious effect into a beneficial trait are discussed.
Measuring the Reduction of the Confinement along the Evolution of a Burn Cut
Authors: Godio Alberto , Seccatore Jacopo
Published: November 2019

We provide an analysis of a tunnel opening cut in a tunnel face. In particular, we focus on the effect of a “burn cut,” which is a typical parallel hole cut. As the evolution of the opening consists of an increase in the volume of the cut prism and the consequent reduction of confinement of the holes along the initiation sequence, we analyze such an evolution using photographic documentation, as well as evaluating the reduction of confinement by measuring the energy transferred from the explosive charge to the rock mass. The energy was estimated by monitoring and analyzing the vibrations induced by the blasting at three different points. We adopted low-cost shock sensors for monitoring the effect of the detonation times at short distance from the blast (about 10 m). The results show an evident reduction of the shock transfer to the rock mass (i.e., a reduction of confinement) with the increase of the cavity opening. Nevertheless, when only the response of the piezometric sensors was considered, a disparity in behavior among the sensors was noted. On the other hand, when the sensor response was integrated over time, the behavior of all sensors normalized, showing the need to focus both on the oscillation and the duration to properly address the shock effect.
Modeling the Effect of Active Modified Atmosphere Packaging on the Microbial Stability and Shelf Life of Gutted Sea Bass
Authors: Tsironi Theofania , Ntzimani Athina , Gogou Eleni , Tsevdou Maria , Semenoglou Ioanna , Dermesonlouoglou Efimia , Taoukis Petros
Published: November 2019

The aim of the study was the evaluation and mathematical modeling of the effect of active modified atmosphere packaging (MAP), by the incorporation of CO2 emitters in the package, on the microbial stability and shelf life of gutted sea bass during refrigerated storage. Gutted sea bass samples were packaged in modified atmosphere (50% CO2–40% N2–10% O2) with and without CO2 emitters (ACT-MAP, MAP) (gas/product volume ratio 3:1) and stored at isothermal conditions: 0 °C, 5 °C, and 10 °C. The gas concentration in the package headspace (%CO2, %O2) and microbial growth (total viable count, TVC, Pseudomonas spp., Enterobacteriaceae spp., lactic acid bacteria) were monitored during storage. The microbial growth was modeled using the Baranyi growth model, and the kinetic parameters (microbial growth rate, lag phase) were estimated at the tested temperature and packaging conditions. The results showed that the ACT-MAP samples presented significantly lower microbial growth compared to the MAP samples. The growth rate of the total viable count at 0 °C was 0.175 and 0.138 d−1 for the MAP and ACT-MAP sea bass, respectively (p < 0.05). The shelf life of the MAP sea bass at 0–10 °C (based on a final TVC value: 7 log CFU g−1) was extended 4–7 days with the addition of a CO2 emitter in the package. The CO2 concentration in the ACT-MAP samples was stabilized at approximately 60%, while the CO2 in the MAP samples was approximately 40% at the end of the shelf life.
A Distributed File-Based Storage System for Improving High Availability of Space Weather Data
Authors: Andrian Yoga , Kim Hyeonwoo , Ju Hongtaek
Published: November 2019

In space science research, the Indonesia National Institute of Aeronautics and Space (LAPAN) is concerned with the development of a system that provides actual information and predictions called the Space Weather Information and Forecast Services (SWIFtS). SWIFtS is supported by a data storage system that serves data, implementing a centralized storage model. This has some problems that impact to researchers as the primary users. The single point of failure and also the delay in data updating on the server is a significant issue when researchers need the latest data, but the server is unable to provide it. To overcome these problems, we proposed a new system that utilized a decentralized model for storing data, leveraging the InterPlanetary File System (IPFS) file system. Our proposed method focused on the automated background process, and its scheme would increase the data availability and throughput by spreading it into nodes through a peer-to-peer connection. Moreover, we also included system monitoring for real-time data flow from each node and information of node status that combines active and passive approaches. For system evaluation, the experiment was performed to determine the performance of the proposed system compared to the existing system by calculating mean replication time and the mean throughput of a node. As expected, performance evaluations showed that our proposed scheme had faster file replication time and supported high throughput.
Construction of Human Behavior Cognitive Map for Robots
Authors: Lin Wei-Zhi , Wang Sui-Hsien , Huang Han-Pang
Published: November 2019

With the advancement of robotics, the importance of service robots in society is increasing. It is crucial for service robots to understand their environment so that they can offer suitable responses to humans. To realize the use of space, robots primarily use an environment model. This paper is focused on the development of an environment model based on human behaviors. In this model, a new neural network structure called dynamic highway networks is applied to recognize humans’ behaviors. In addition, a two-dimensional pose estimator, Laban movement analysis, and the fuzzy integral are employed. With these methods, two new behavior-recognition algorithms are developed, and a method to record the relationship between behavior and environment is proposed. Based on the proposed environmental model, robots can identify abnormal behavior, provide an appropriate response and guide a person toward the desired normal behavior by identifying abnormal behavior. Simulations and experiments justify the proposed method with satisfactory results.
Developing Innovative Crutch Using IDeS (Industrial Design Structure) Methodology
Authors: Frizziero Leonardo , Donnici Giampiero , Liverani Alfredo , Alessandri Giulia , Menozzi Grazia Chiara , Varotti Emma
Published: November 2019

The present study wants to bring to light a new type of crutch designed for a chronic patient with perennial limited mobility, who must use this support every time a move is needed. The main purpose of the project consists in recommending a correct use of the crutch through technology, limiting the damage normally caused by a bad use of crutches and giving a support both for the patient and for the doctor. All of the features of the crutches were defined through relationship matrices and a benchmarking, which helped us for defining the requirements; other important features were defined, taking a look to the technological progresses applied to new, patented crutches. The result is a sensorized crutch, functional and oriented to meet the user’s needs in order to prevent an incorrect use of the support avoiding the growth of other pains.
Gray Relational Analysis Optimization for Coalbed Methane Blocks in Complex Conditions Based on a Best Worst and Entropy Method
Authors: Zhao Guofei , Kang Tianhe , Guo Junqing , Zhang Runxu , Li Ligong
Published: November 2019

The uncertainty and complexity of the geological parameters of coalbed methane (CBM) reservoirs may result in the assignment of atypical exact values to a whole block. Therefore, the influence of different resources and productivity characterization indices on the development of CBM blocks was analyzed systematically in this research. Furthermore, this research reported an interval-number index evaluation system and a gray relational analysis optimization model for CBM development. The weights of the evaluation indices were determined by combining the entropy method and the best worst method (BWM) with the objective deviation law of the data and the subjective experience of experts. Based on the gray system theory, a multi-index gray relational analysis optimization model was established to evaluate the interval values and optimize the development potential of CBM blocks with complex geological conditions. The results show that the order of importance of the eight resources and reservoir evaluation indices, from highest to lowest, is permeability, reservoir pressure gradient, gas content, reservoir thickness, reservoir depth, critical reservoir ratio, Young’s modulus, and reservoir temperature. The order of the target blocks, from most to least optimal, is the Sanjiao block, Fanzhuang-Zhengzhuang block, Daning-Jixian block, Gujiao block, and Fengrun block. The evaluation results are consistent with actual conditions. Thus, the multi-index gray relation analysis optimization model, on account of the gray system theory, is feasible for the development evaluation of CBM blocks under complex conditions. This model is suitable for the CBM blocks in the range of 547 km2~675 km2. When using this model, the index values must be representative and accurate.
Ni3Se4@MoSe2 Composites for Hydrogen Evolution Reaction
Authors: Guo Wenwu , Le Quyet Van , Do Ha Huu , Hasani Amirhossein , Tekalgne Mahider , Bae Sa-Rang , Lee Tae Hyung , Jang Ho Won , Ahn Sang Hyun , Kim Soo Young
Published: November 2019

Transition metal dichalcogenides (TMDs) have been considered as one of the most promising electrocatalysts for the hydrogen evolution reaction (HER). Many studies have demonstrated the feasibility of significant HER performance improvement of TMDs by constructing composite materials with Ni-based compounds. In this work, we prepared Ni3Se4@MoSe2 composites as electrocatalysts for the HER by growing in situ MoSe2 on the surface of Ni3Se4 nanosheets. Electrochemical measurements revealed that Ni3Se4@MoSe2 nanohybrids are highly active and durable during the HER process, which exhibits a low onset overpotential (145 mV) and Tafel slope (65 mV/dec), resulting in enhanced HER performance compared to pristine MoSe2 nanosheets. The enhanced HER catalytic activity is ascribed to the high surface area of Ni3Se4 nanosheets, which can both efficiently prevent the agglomeration issue of MoSe2 nanosheets and create more catalytic edge sites, hence accelerate electron transfer between MoSe2 and the working electrode in the HER. This approach provides an effective pathway for catalytic enhancement of MoSe2 electrocatalysts and can be applied for other TMD electrocatalysts.
Analysis and Development of Hybrid Earphone Combining Balanced-Armature and Dynamic Receivers
Authors: Jiang Yuan-Wu , Xu Dan-Ping , Jiang Zhi-Xiong , Kim Jun-Hyung , Hwang Sang-Moon
Published: November 2019

With the rapid progress in the development of multimedia devices, earphones have become increasingly important as audio output tools. Hybrid earphones combining balanced-armature (BA) and dynamic receivers can produce better performance over a wider range when compared to the earphones with BA receiver alone (BA earphones) or dynamic receiver alone (dynamic earphones). BA and dynamic earphones are multi-physics products that exhibit coupling between the electromagnetic, mechanical, and acoustic domains. In this study, an analysis tool is developed to design a hybrid earphone based on the conventional BA and dynamic earphones. Using the developed analysis tool, an acoustic tube is optimized to match the earphone target curve and obtain improved sound quality. A prototype is manufactured and tested, and the experimental results verify the feasibility and effectiveness of the developed analysis tool. The root-mean-square value of the sound pressure level (SPL) deviation of the hybrid earphone with the optimized acoustic tube is 4.60, whereas those for the dynamic and BA earphones are 8.94 and 6.04, respectively. Thus, it is verified that the frequency response is improved using the hybrid earphone developed herein.
Parameterization of The Single-Scattering Properties of Dust Aerosols in Radiative Flux Calculations
Authors: Wang Meihua , Su Jing , Li Xugang , Wang Chen , Ge Jinming
Published: November 2019

In this study, we present parameterization schemes of dust single-scattering properties (SSPs) in order to establish a fast and accurate way to obtain the SSPs for dust shortwave radiative flux calculation. Based on the assumption that dust particles are spheroids, we represent a single nonspherical particle with a collection of monodisperse spheres that contain the same total surface area and volume as the original particle to convert the spheroid to a sphere. The SSPs of dust particles were parameterized in terms of the effective radius ( R e ) and imaginary part of the refractive index ( M i ). The averaged relative errors of the parameterized to the “exact” single-scattering properties, which refer to the results from the Mie theory program, are below 1.5%. To further quantify the impact of parametrization on the radiative flux simulation, we computed the radiative fluxes at both the top of the atmosphere (TOA) and the surface by using SSPs from the parameterization and the “exact”, respectively. The maximum relative errors were below 1% at both the TOA and the surface, proving that the SSPs of dust calculated by our parameterization schemes are well suited for radiative flux calculations. This parameterization differs from previous works by being formulated not only with R e but also with M i . We also investigated the sensitivity of dust-aerosol forcing to R e , M i , optical depth (τ), and solar zenith angle (SZA). The results show that the value of shortwave (SW) radiative forcing (RF) at the TOA changes from negative to positive as the M i is increasing, which means that, as the absorption of dust particles becomes stronger, more energy is kept in the atmosphere to heat the earth–atmosphere system. The SW RF gradually becomes less negative at the TOA and more negative at the surface with increasing R e , due to the decreases of reflection and transmission along with the single-scattering albedo decreasing. As the optical depth increases, the values of the SW RF decrease because of the strong attenuation for heavy loading. When SZA increases, the SW RF becomes more negative at both the TOA and the surface due to the long optical path at a large SZA. The errors induced from the parameterized SSPs of dust in the SW RF calculation are very small, which are less than 2.1%, demonstrating the accuracy of the parameterization and its reliability for climate model applications.
Urban Areas and Urban–Rural Contrasts under Climate Change: What Does the EURO-CORDEX Ensemble Tell Us?—Investigating near Surface Humidity in Berlin and Its Surroundings
Authors: Langendijk Gaby S. , Rechid Diana , Jacob Daniela
Published: November 2019

Climate change will impact urban areas. Decision makers need useful climate information to adapt adequately. This research aims to improve understanding of changes in moisture and temperature projected under climate change in Berlin compared to its surroundings. Simulations for the Representative Concentration Pathway (RCP) 8.5 scenario from the European Coordinated Regional Climate Downscaling Experiment (EURO-CORDEX) 0.11° are analyzed, showing a difference in moisture and temperature variables between Berlin and its surroundings. The running mean over 30 years shows a divergence throughout the twenty-first century for relative humidity between Berlin and its surroundings. Under this scenario, Berlin gets drier over time. The Mann-Kendall test quantifies a robust decreasing trend in relative humidity for the multi-model ensemble throughout the twenty-first century. The Mann-Whitney-Wilcoxon test for relative humidity indicates a robust climate change signal in Berlin. It is drier and warmer in Berlin compared to its surroundings for all months with the largest difference existing in summer. Additionally, the change in humidity for the period 2070–2099 compared to 1971–2000 is larger in the summer months. This study presents results to better understand near surface moisture change and related variables under long-term climate change in urban areas compared to their rural surroundings using a regional climate multi-model ensemble.
The Influence of Tornado Activity, Impact, Memory, and Sentiment on Tornado Perception Accuracy among College Students
Authors: Senkbeil Jason C. , Ellis Kelsey N. , Reed Jacob R.
Published: November 2019

A survey consisting of open-ended and closed responses was administered at three universities in the eastern USA. The home counties of survey participants represented climatological tornado risks spanning from rarely impacted to frequently impacted. The first objective of this research was to classify climatological tornado risk for each county so that analyses of tornado perception accuracy could be evaluated. Perception accuracy was defined as the difference between what each participant perceived minus what actually happened. A manual classification scheme was created that uses the Storm Prediction Center’s Convective Outlook framework as county climatological risk categories. Participants from high-risk counties statistically significantly overestimated the numbers of violent tornadoes compared to participants from every risk category but moderate. Furthermore, participants from high-risk counties had significantly greater tornado impacts, thus validating the classification of high-risk. Participants from high, moderate, and slight-risk counties significantly overestimated the number of strong tornadoes compared to participants from enhanced-risk counties. There appeared to be no relationships between tornado memory and tornado sentiment with tornado perception accuracy. Possible explanations for the overestimation of the numbers of violent tornadoes in high-risk counties are discussed.
Projections of Temperature-Attributable Deaths in Portuguese Metropolitan Areas: A Time-Series Modelling Approach
Authors: Rodrigues Mónica , Santana Paula , Rocha Alfredo
Published: November 2019

Climate change is now widely recognised as the greatest global threat over the coming decades. This study aimed to quantify and project the effects of climate change on future temperature-attributable mortality due to circulatory system diseases (CSD) in Lisbon metropolitan area (LMA) and in Porto metropolitan area (PMA). The future time slices of Representative Concentration Pathway (RCP 8.5), mid-term (2046–2065) and long-term (2080–2099) were compared with the reference period (1986–2005). There is a significant decreasing trend in proportion to the overall extreme cold temperature-attributable mortality due to CSD in the future periods (2045–2065 and 2081–2099) in LMA, −0.63% and −0.73%, respectively, and in PMA, −0.62% for 2045–2065 and −0.69% for 2081–2099, compared to the historical period. The fraction attributable to extreme hot temperature in the summer months increased by 0.08% and 0.23%, from 0.04% in the historical period to 0.11% during 2046–2065, and to 0.27% during 2081–2099 in LMA. While there were no noticeable changes due to extreme hot temperature during the summer in PMA, significant increases were observed with warmer winter temperatures: 1.27% and 2.80%. The projections of future temperature-attributable mortality may provide valuable information to support climate policy decision making and temperature-related risk management.
Evolutionary Conservation and Expression Patterns of Neutral/Alkaline Invertases in Solanum
Authors: Pan Luzhao , Guo Qinwei , Chai Songlin , Cheng Yuan , Ruan Meiying , Ye Qingjing , Wang Rongqing , Yao Zhuping , Zhou Guozhi , Li Zhimiao , Deng Minghua , Jin Fengmei , Liu Lecheng , Wan Hongjian
Published: November 2019

The invertase gene family in plants is composed of two subfamilies of enzymes, namely, acid- and neutral/alkaline invertases (cytosolic invertase, CIN). Both can irreversibly cleave sucrose into fructose and glucose, which are thought to play key roles in carbon metabolism and plant growth. CINs are widely found in plants, but little is reported about this family. In this paper, a comparative genomic approach was used to analyze the CIN gene family in Solanum, including Solanum tuberosum, Solanum lycopersicum, Solanum pennellii, Solanum pimpinellifolium, and Solanum melongena. A total of 40 CINs were identified in five Solanum plants, and sequence features, phylogenetic relationships, motif compositions, gene structure, collinear relationship, and expression profile were further analyzed. Sequence analysis revealed a remarkable conservation of CINs in sequence length, gene number, and molecular weight. The previously verified four amino acid residues (D188, E414, Arg430, and Ser547) were also observed in 39 out of 40 CINs in our study, showing to be deeply conserved. The CIN gene family could be distinguished into groups α and β, and α is further subdivided into subgroups α1 and α2 in our phylogenetic tree. More remarkably, each species has an average of four CINs in the α and β groups. Marked interspecies conservation and collinearity of CINs were also further revealed by chromosome mapping. Exon–intron configuration and conserved motifs were consistent in each of these α and β groups on the basis of in silico analysis. Expression analysis indicated that CINs were constitutively expressed and share similar expression profiles in all tested samples from S. tuberosum and S. lycopersicum. In addition, in CIN genes of the tomato and potato in response to abiotic and biotic stresses, phytohormones also performed. Overall, CINs in Solanum were encoded by a small and highly conserved gene family, possibly reflecting structural and functional conservation in Solanum. These results lay the foundation for further expounding the functional characterization of CIN genes and are also significant for understanding the evolutionary profiling of the CIN gene family in Solanum.
The Plasma Factor XIII Heterotetrameric Complex Structure: Unexpected Unequal Pairing within a Symmetric Complex
Authors: Singh Sneha , Nazabal Alexis , Kaniyappan Senthilvelrajan , Pellequer Jean-Luc , Wolberg Alisa S. , Imhof Diana , Oldenburg Johannes , Biswas Arijit
Published: November 2019

Factor XIII (FXIII) is a predominant determinant of clot stability, strength, and composition. Plasma FXIII circulates as a pro-transglutaminase with two catalytic A subunits and two carrier-protective B subunits in a heterotetramer (FXIII-A2B2). FXIII-A2 and -B2 subunits are synthesized separately and then assembled in plasma. Following proteolytic activation by thrombin and calcium-mediated dissociation of the B subunits, activated FXIII (FXIIIa) covalently cross links fibrin, promoting clot stability. The zymogen and active states of the FXIII-A subunits have been structurally characterized; however, the structure of FXIII-B subunits and the FXIII-A2B2 complex have remained elusive. Using integrative hybrid approaches including atomic force microscopy, cross-linking mass spectrometry, and computational approaches, we have constructed the first all-atom model of the FXIII-A2B2 complex. We also used molecular dynamics simulations in combination with isothermal titration calorimetry to characterize FXIII-A2B2 assembly, activation, and dissociation. Our data reveal unequal pairing of individual subunit monomers in an otherwise symmetric complex, and suggest this unusual structure is critical for both assembly and activation of this complex. Our findings enhance understanding of mechanisms associating FXIII-A2B2 mutations with disease and have important implications for the rational design of molecules to alter FXIII assembly or activity to reduce bleeding and thrombotic complications.
Salivary Protein Panel to Diagnose Systolic Heart Failure
Authors: Zhang Xi , Broszczak Daniel , Kostner Karam , Guppy-Coles Kristyan B , Atherton John J , Punyadeera Chamindie
Published: November 2019

Screening for systolic heart failure (SHF) has been problematic. Heart failure management guidelines suggest screening for structural heart disease and SHF prevention strategies should be a top priority. We developed a multi-protein biomarker panel using saliva as a diagnostic medium to discriminate SHF patients and healthy controls. We collected saliva samples from healthy controls (n = 88) and from SHF patients (n = 100). We developed enzyme linked immunosorbent assays to quantify three specific proteins/peptide (Kallikrein-1, Protein S100-A7, and Cathelicidin antimicrobial peptide) in saliva samples. The analytical and clinical performances and predictive value of the proteins were evaluated. The analytical performances of the immunoassays were all within acceptable analytical ranges. The multi-protein panel was able to significantly (p < 0.001) discriminate saliva samples collected from patients with SHF from controls. The multi-protein panel demonstrated good performance with an overall diagnostic accuracy of 81.6% (sensitivity of 79.2% and specificity of 85.7%) when distinguishing SHF patients from healthy individuals. In conclusion, we have developed immunoassays to measure the salivary concentrations of three proteins combined as a panel to accurately distinguish SHF patients from healthy controls. While this requires confirmation in larger cohorts, our findings suggest that this three-protein panel has the potential to be used as a biomarker for early detection of SHF.
Identification, Classification and Screening for γ-Amino-butyric Acid Production in Lactic Acid Bacteria from Cambodian Fermented Foods
Authors: Ly Dalin , Mayrhofer Sigrid , Agung Yogeswara I. , Nguyen Thu-Ha , Domig Konrad
Published: November 2019

Screening for various types of lactic acid bacteria (LAB) that form the biological agent γ-amino-butyric acid (GABA) is important to produce different kinds of GABA-containing fermented foods. So far, no GABA-producing LAB have been reported from Cambodian fermented foods. Most small-scale fermentations and even some industrial processes in this country still rely on indigenous LAB. The application of GABA-producing autochthonous starters would allow the production of Cambodian fermented foods with an additional nutritional value that meet the population’s dietary habits and that are also more attractive for the international food market. Matrix-assisted laser desorption/ionizing time-of-flight mass spectrometry (MALDI-TOF MS) and partial 16S rDNA sequencing were used to identify 68 LAB isolates from Cambodian fermented foods. These isolates were classified and grouped with (GTG)5 rep-PCR, resulting in 50 strains. Subsequently, all strains were investigated for their ability to produce GABA by thin layer chromatography. GABA-positive strains were further analyzed by the GABase assay. Of the six GABA-positive LAB strains—one Lactobacillus futsaii, two Lactobacillus namurensis, and three Lactobacillus plantarum strains—two Lactobacillus plantarum strains produced high amounts of GABA (20.34 mM, 16.47 mM). These strains should be further investigated for their potential application as GABA-producing starter cultures in the food applications.
The Role of 18F-FDG PET/CT in Staging and Prognostication of Mantle Cell Lymphoma: An Italian Multicentric Study
Authors: Albano Domenico , Laudicella Riccardo , Ferro Paola , Allocca Michela , Abenavoli Elisabetta , Buschiazzo Ambra , Castellino Alessia , Chiaravalloti Agostino , Cuccaro Annarosa , Cuppari Lea , Durmo Rexhep , Evangelista Laura , Frantellizzi Viviana , Kovalchuk Sofya , Linguanti Flavia , Santo Giulia , Bauckneht Matteo , Annunziata Salvatore
Published: November 2019

Mantle cell lymphoma (MCL) is an aggressive lymphoma subtype with poor prognosis in which 18F-FDG-PET/CT role in treatment response evaluation and prediction of outcome is still unclear. The aim of this multicentric study was to investigate the role of 18F-FDG-PET/CT in staging MCL and the prognostic role of Deauville criteria (DC) in terms of progression-free survival (PFS) and overall survival (OS). We retrospectively enrolled 229 patients who underwent baseline and end-of-treatment (eot) 18F-FDG-PET/CT after first-line therapy. EotPET/CT scans were visually interpreted according to DC. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of PET/CT for evaluation of bone marrow (BM) were 27%, 100%, 100%, 48% and 57%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of PET/CT for evaluation of the gastrointestinal (GI) tract were 60%, 99%, 93%, 90% and 91%, respectively. At a median follow-up of 40 months, relapse occurred in 104 cases and death in 49. EotPET/CT results using DC significantly correlated with PFS, not with OS. Instead, considering OS, only MIPI score was significantly correlated. In conclusion, we demonstrated that MCL is an FDG-avid lymphoma and 18F-FDG-PET/CT is a useful tool for staging purpose, showing good specificity for BM and GI evaluation, but suboptimal sensitivity. EotPET/CT result was the only independent significant prognostic factor that correlated with PFS.
JAK/STAT-Activating Genomic Alterations Are a Hallmark of T-PLL
Authors: Wahnschaffe Linus , Braun Till , Timonen Sanna , Giri Anil K. , Schrader Alexandra , Wagle Prerana , Almusa Henrikki , Johansson Patricia , Bellanger Dorine , López Cristina , Haferlach Claudia , Stern Marc-Henri , Dürig Jan , Siebert Reiner , Mustjoki Satu , Aittokallio Tero , Herling Marco
Published: November 2019

T-cell prolymphocytic leukemia (T-PLL) is a rare and poor-prognostic mature T-cell leukemia. Recent studies detected genomic aberrations affecting JAK and STAT genes in T-PLL. Due to the limited number of primary patient samples available, genomic analyses of the JAK/STAT pathway have been performed in rather small cohorts. Therefore, we conducted—via a primary-data based pipeline—a meta-analysis that re-evaluated the genomic landscape of T-PLL. It included all available data sets with sequence information on JAK or STAT gene loci in 275 T-PLL. We eliminated overlapping cases and determined a cumulative rate of 62.1% of cases with mutated JAK or STAT genes. Most frequently, JAK1 (6.3%), JAK3 (36.4%), and STAT5B (18.8%) carried somatic single-nucleotide variants (SNVs), with missense mutations in the SH2 or pseudokinase domains as most prevalent. Importantly, these lesions were predominantly subclonal. We did not detect any strong association between mutations of a JAK or STAT gene with clinical characteristics. Irrespective of the presence of gain-of-function (GOF) SNVs, basal phosphorylation of STAT5B was elevated in all analyzed T-PLL. Fittingly, a significant proportion of genes encoding for potential negative regulators of STAT5B showed genomic losses (in 71.4% of T-PLL in total, in 68.4% of T-PLL without any JAK or STAT mutations). They included DUSP4, CD45, TCPTP, SHP1, SOCS1, SOCS3, and HDAC9. Overall, considering such losses of negative regulators and the GOF mutations in JAK and STAT genes, a total of 89.8% of T-PLL revealed a genomic aberration potentially explaining enhanced STAT5B activity. In essence, we present a comprehensive meta-analysis on the highly prevalent genomic lesions that affect genes encoding JAK/STAT signaling components. This provides an overview of possible modes of activation of this pathway in a large cohort of T-PLL. In light of new advances in JAK/STAT inhibitor development, we also outline translational contexts for harnessing active JAK/STAT signaling, which has emerged as a ‘secondary’ hallmark of T-PLL.
Changes of O6-Methylguanine DNA Methyltransferase (MGMT) Promoter Methylation in Glioblastoma Relapse—A Meta-Analysis Type Literature Review
Authors: Feldheim Jonas , Kessler Almuth F. , Monoranu Camelia M. , Ernestus Ralf-Ingo , Löhr Mario , Hagemann Carsten
Published: November 2019

Methylation of the O6-methylguanine DNA methyltransferase (MGMT) promoter has emerged as strong prognostic factor in the therapy of glioblastoma multiforme. It is associated with an improved response to chemotherapy with temozolomide and longer overall survival. MGMT promoter methylation has implications for the clinical course of patients. In recent years, there have been observations of patients changing their MGMT promoter methylation from primary tumor to relapse. Still, data on this topic are scarce. Studies often consist of only few patients and provide rather contrasting results, making it hard to draw a clear conclusion on clinical implications. Here, we summarize the previous publications on this topic, add new cases of changing MGMT status in relapse and finally combine all reports of more than ten patients in a statistical analysis based on the Wilson score interval. MGMT promoter methylation changes are seen in 115 of 476 analyzed patients (24%; CI: 0.21–0.28). We discuss potential reasons like technical issues, intratumoral heterogeneity and selective pressure of therapy. The clinical implications are still ambiguous and do not yet support a change in clinical practice. However, retesting MGMT methylation might be useful for future treatment decisions and we encourage clinical studies to address this topic.
Reversal of Epithelial–Mesenchymal Transition by Natural Anti-Inflammatory and Pro-Resolving Lipids
Authors: Lee Chang Hoon
Published: November 2019

Epithelial mesenchymal transition (EMT) is a key process in the progression of malignant cancer. Therefore, blocking the EMT can be a critical fast track for the development of anticancer drugs. In this paper, we update recent research output of EMT and we explore suppression of EMT by natural anti-inflammatory compounds and pro-resolving lipids.
Mutations in Splicing Factor Genes in Myeloid Malignancies: Significance and Impact on Clinical Features
Authors: Visconte Valeria , O. Nakashima Megan , J. Rogers Heesun
Published: November 2019

Components of the pre-messenger RNA splicing machinery are frequently mutated in myeloid malignancies. Mutations in LUC7L2, PRPF8, SF3B1, SRSF2, U2AF1, and ZRSR2 genes occur at various frequencies ranging between 40% and 85% in different subtypes of myelodysplastic syndrome (MDS) and 5% and 10% of acute myeloid leukemia (AML) and myeloproliferative neoplasms (MPNs). In some instances, splicing factor (SF) mutations have provided diagnostic utility and information on clinical outcomes as exemplified by SF3B1 mutations associated with increased ring sideroblasts (RS) in MDS-RS or MDS/MPN-RS with thrombocytosis. SF3B1 mutations are associated with better survival outcomes, while SRSF2 mutations are associated with a shorter survival time and increased AML progression, and U2AF1 mutations with a lower remission rate and shorter survival time. Beside the presence of mutations, transcriptomics technologies have shown that one third of genes in AML patients are differentially expressed, leading to altered transcript stability, interruption of protein function, and improper translation compared to those of healthy individuals. The detection of SF mutations demonstrates the importance of splicing abnormalities in the hematopoiesis of MDS and AML patients given the fact that abnormal splicing regulates the function of several transcriptional factors (PU.1, RUNX1, etc.) crucial in hematopoietic function. This review provides a summary of the significance of the most frequently mutated SF genes in myeloid malignancies and an update on novel targeted therapies in experimental and clinical trial stages.
Targeting Angiogenesis by Blocking the ATM–SerRS–VEGFA Pathway for UV-Induced Skin Photodamage and Melanoma Growth
Authors: Song Yadong , Lu Hongyan , Wang Qiong , Xiang Rong
Published: November 2019

Retinoic acid (RA) has been widely used to protect skin from photo damage and skin carcinomas caused by solar ultraviolet (UV) irradiation, yet the mechanism remains elusive. Here, we report that all-trans retinoic acid (tRA) can directly induce the expression of a newly identified potent anti-angiogenic factor, seryl tRNA synthetase (SerRS), whose angiostatic role can, however, be inhibited by UV-activated ataxia telangiectasia mutated (ATM) kinase. In both a human epidermal cell line, HaCaT, and a mouse melanoma B16F10 cell line, we found that tRA could activate SerRS transcription through binding with the SerRS promoter. However, UV irradiation induced activation of ATM-phosphorylated SerRS, leading to the inactivation of SerRS as a transcriptional repressor of vascular endothelial growth factor A (VEGFA), which dampened the effect of tRA. When combined with ATM inhibitor KU-55933, tRA showed a greatly enhanced efficiency in inhibiting VEGFA expression and a much better protection of mouse skin from photo damage. Also, we found the combination greatly inhibited tumor angiogenesis and growth in mouse melanoma xenograft in vivo. Taken together, tRA combined with an ATM inhibitor can greatly enhance the anti-angiogenic activity of SerRS under UV irradiation and could be a better strategy in protecting skin from angiogenesis-associated skin damage and melanoma caused by UV radiation.
The Potential Impact of Connexin 43 Expression on Bcl-2 Protein Level and Taxane Sensitivity in Head and Neck Cancers–In Vitro Studies
Authors: Gurbi Bianka , Brauswetter Diána , Varga Attila , Gyulavári Pál , Pénzes Kinga , Murányi József , Zámbó Veronika , Birtalan Ede , Krenács Tibor , Becker David Laurence , Csala Miklós , Vályi-Nagy István , Peták István , Dános Kornél
Published: November 2019

The poor prognosis of head and neck squamous cell carcinoma (HNSCC) is partly due to the lack of reliable predictive markers. Connexin 43 (Cx43) protein and its cell-communication channels have been assigned tumor suppressor functions while the anti-apoptotic Bcl-2 (B-cell lymphoma-2) protein has been associated with negative prognostic significance in cancer. This study aimed to test the role of Cx43 protein on Bcl-2 expression, tumor progression and response to taxane-based treatment in HNSCC. Human papillomavirus (HPV) negative HNSCC cell lines were tested for paclitaxel sensitivity through measuring apoptosis induction, cell viability and changes in Cx43 and Bcl-2 levels using flow cytometry, cell viability assay, immunocytochemistry and western blot. Inhibition of Cx43 expression using siRNA increased Bcl-2 protein levels in SCC25 (tongue squamous cell carcinoma) cells, while forced Cx43 expression reduced Bcl-2 levels and supported paclitaxel cytotoxicity in FaDu (hypopharynx squamous cell carcinoma) cells. In vitro results were in line with protein expression and clinicopathological features tested in tissue microarray samples of HNSCC patients. Our data demonstrate that elevated Cx43 and reduced Bcl-2 levels may indicate HNSCC sensitivity to taxane-based treatments. On the contrary, silencing of the Cx43 gene GJA1 (gap junction protein alpha-1) can result in increased Bcl-2 expression and reduced paclitaxel efficiency. Clinical tumor-based analysis also confirmed the inverse correlation between Cx43 and Bcl-2 expression.
The Protein Neddylation Inhibitor MLN4924 Suppresses Patient-Derived Glioblastoma Cells via Inhibition of ERK and AKT Signaling
Authors: Han Suji , Shin Hyemi , Oh Jeong-Woo , Oh Yun Jeong , Her Nam-Gu , Nam Do-Hyun
Published: November 2019

Glioblastoma is a highly aggressive and lethal brain tumor, with limited treatment options. Abnormal activation of the neddylation pathway is observed in glioblastoma, and the NEDD8-activating enzyme (NAE) inhibitor, MLN4924, was previously shown to be effective in glioblastoma cell line models. However, its effect has not been tested in patient-derived glioblastoma stem cells. We first analyzed public data to determine whether NEDD8 pathway proteins are important in glioblastoma development and patient survival. NAE1 and UBA3 levels increased in glioblastoma patients; high NEDD8 levels were associated with poor clinical outcomes. Immunohistochemistry results also supported this result. The effects of MLN4924 were evaluated in 4 glioblastoma cell lines and 15 patient-derived glioblastoma stem cells using high content analysis. Glioblastoma cell lines and patient-derived stem cells were highly susceptible to MLN4924, while normal human astrocytes were resistant. In addition, there were various responses in 15 patient-derived glioblastoma stem cells upon MLN4924 treatment. Genomic analyses indicated that MLN4924 sensitive cells exhibited enrichment of Extracellular Signal Regulated Kinase (ERK) and Protein kinase B (AKT, also known as PKB) signaling. We verified that MLN4924 inhibits ERK and AKT phosphorylation in MLN4924 sensitive cells. Our findings suggest that patient-derived glioblastoma stem cells in the context of ERK and AKT activation are sensitive and highly regulated by neddylation inhibition.