bluuebunny/crossref_metadata_2025 huggingface dataset:
Components from the Human c-myb Transcriptional Regulation System Reactivate Epigenetically Repressed Transgenes
Authors: Barrett Cassandra M. , McCracken Reilly , Elmer Jacob , Haynes Karmella A.
Published: January 2020

A persistent challenge for mammalian cell engineering is the undesirable epigenetic silencing of transgenes. Foreign DNA can be incorporated into closed chromatin before and after it has been integrated into a host cell’s genome. To identify elements that mitigate epigenetic silencing, we tested components from the c-myb and NF-kB transcriptional regulation systems in transiently transfected DNA and at chromosomally integrated transgenes in PC-3 and HEK 293 cells. DNA binding sites for MYB (c-myb) placed upstream of a minimal promoter enhanced expression from transiently transfected plasmid DNA. We targeted p65 and MYB fusion proteins to a chromosomal transgene, UAS-Tk-luciferase, that was silenced by ectopic Polycomb chromatin complexes. Transient expression of Gal4-MYB induced an activated state that resisted complete re-silencing. We used custom guide RNAs and dCas9-MYB to target MYB to different positions relative to the promoter and observed that transgene activation within ectopic Polycomb chromatin required proximity of dCas9-MYB to the transcriptional start site. Our report demonstrates the use of MYB in the context of the CRISPR-activation system, showing that DNA elements and fusion proteins derived from c-myb can mitigate epigenetic silencing to improve transgene expression in engineered cell lines.
Inhibitory Effects of Peptide Lunasin in Colorectal Cancer HCT-116 Cells and Their Tumorsphere-Derived Subpopulation
Authors: Fernández-Tomé Samuel , Xu Fei , Han Yanhui , Hernández-Ledesma Blanca , Xiao Hang
Published: January 2020

The involvement of cancer stem-like cells (CSC) in the tumor pathogenesis has profound implications for cancer therapy and chemoprevention. Lunasin is a bioactive peptide from soybean and other vegetal sources with proven protective activities against cancer and other chronic diseases. The present study focused on the cytotoxic effect of peptide lunasin in colorectal cancer HCT-116 cells, both the bulk tumor and the CSC subpopulations. Lunasin inhibited the proliferation and the tumorsphere-forming capacity of HCT-116 cells. Flow cytometry results demonstrated that the inhibitory effects were related to apoptosis induction and cell cycle-arrest at G1 phase. Moreover, lunasin caused an increase in the sub-GO/G1 phase of bulk tumor cells, linked to the apoptotic events found. Immunoblotting analysis further showed that lunasin induced apoptosis through activation of caspase-3 and cleavage of PARP, and could modulate cell cycle progress through the cyclin-dependent kinase inhibitor p21. Together, these results provide new evidence on the chemopreventive activity of peptide lunasin on colorectal cancer by modulating both the parental and the tumorsphere-derived subsets of HCT-116 cells.
A Sub-Clone of RAW264.7-Cells Form Osteoclast-Like Cells Capable of Bone Resorption Faster than Parental RAW264.7 through Increased De Novo Expression and Nuclear Translocation of NFATc1
Authors: Mira-Pascual Laia , Tran Anh N. , Andersson Göran , Näreoja Tuomas , Lång Pernilla
Published: January 2020

The murine macrophage cell line RAW264.7 is extensively used as a progenitor to study osteoclast (OC) differentiation. RAW264.7 is a heterogeneous cell line, containing sub-clones with different abilities to form OCs. The aim of this study was to identify characteristics within the heterogeneous RAW264.7 cells that define sub-clones with an augmented ability to form bone-resorbing OCs (H9), as well as sub-clones representing non-OCs (J8). RAW264.7 sub-clones were isolated by single cell cloning. Selection was based on TRAP/cathepsin K expression in sub-clone cultures without added RANKL. Sub-clones before and after differentiation with RANKL were assayed for multiple OC-characteristics. Sub-clone H9 cells presented a higher expression of OC-markers in cultures without added RANKL compared to the parental RAW264.7. After 6 days of RANKL stimulation, sub-clone H9 cells had equal expression levels of OC-markers with RAW264.7 and formed OCs able to demineralize hydroxyapatite. However, sub-clone H9 cells displayed rapid differentiation of OC already at Day 2 compared to Day 4 from parental RAW264.7, and when cultured on plastic and on bone they were more efficient in resorption. This rapid differentiation was likely due to high initial expression/nuclear translocation of OC master transcription factor, NFATc1. In contrast to H9, J8 cells expressed initially very low levels of OC-markers, and they did not respond to RANKL-stimulation by developing OC-characteristics/OC-marker expression. Hence, H9 is an additional clone suitable for experimental setup requiring rapid differentiation of large numbers of OCs.
Genetic Dissection of Grain Yield and Agronomic Traits in Maize under Optimum and Low-Nitrogen Stressed Environments
Authors: Tadesse Ertiro Berhanu , Olsen Michael , Das Biswanath , Gowda Manje , Labuschagne Maryke
Published: January 2020

Understanding the genetic basis of maize grain yield and other traits under low-nitrogen (N) stressed environments could improve selection efficiency. In this study, five doubled haploid (DH) populations were evaluated under optimum and N-stressed conditions, during the main rainy season and off-season in Kenya and Rwanda, from 2014 to 2015. Identifying the genomic regions associated with grain yield (GY), anthesis date (AD), anthesis-silking interval (ASI), plant height (PH), ear height (EH), ear position (EPO), and leaf senescence (SEN) under optimum and N-stressed environments could facilitate the use of marker-assisted selection to develop N-use-efficient maize varieties. DH lines were genotyped with genotyping by sequencing. A total of 13, 43, 13, 25, 30, 21, and 10 QTL were identified for GY, AD ASI, PH, EH, EPO, and SEN, respectively. For GY, PH, EH, and SEN, the highest number of QTL was found under low-N environments. No common QTL between optimum and low-N stressed conditions were identified for GY and ASI. For secondary traits, there were some common QTL for optimum and low-N conditions. Most QTL conferring tolerance to N stress was on a different chromosome position under optimum conditions.
Recursive Matrix Calculation Paradigm by the Example of Structured Matrix
Authors: Respondek Jerzy S.
Published: January 2020

In this paper, we derive recursive algorithms for calculating the determinant and inverse of the generalized Vandermonde matrix. The main advantage of the recursive algorithms is the fact that the computational complexity of the presented algorithm is better than calculating the determinant and the inverse by means of classical methods, developed for the general matrices. The results of this article do not require any symbolic calculations and, therefore, can be performed by a numerical algorithm implemented in a specialized (like Matlab or Mathematica) or general-purpose programming language (C, C++, Java, Pascal, Fortran, etc.).
QFR Predicts the Incidence of Long-Term Adverse Events in Patients with Suspected CAD: Feasibility and Reproducibility of the Method
Authors: Buono Andrea , Mühlenhaus Annika , Schäfer Tabitha , Trieb Ann-Kristin , Schmeißer Julian , Koppe Franziska , Münzel Thomas , Anadol Remzi , Gori Tommaso
Published: January 2020

AIMS: We evaluate feasibility and reproducibility of post hoc quantitative flow ratio (QFR) measurements and their prognostic predictive power during long-term follow-up. METHODS AND RESULTS: Between 2010 and 2012, 167 patients without angiographic evidence of significant stenoses were enrolled in a prospective registry. Of these patients, 96% presented 7 years follow-up data. QFR was measured post hoc by three certified investigators. QFR analysis was feasible in 71% of left anterior descending (LAD), 72% of left circumflex (LCX), and 61% of right (RCA) coronaries for a total of 350 measurements repeated in triplicate. Coefficients of variation were 2.1% for RCA and LCX, and 2.8% for the LAD (quartile coefficients of dispersion respectively 1.5, 1.4, and 1.3). QFR ≤0.80 was recorded in 25 patients (27 vessels, in 74% of the cases LAD). A total of 86 major adverse cardiovascular and cerebrovascular events were observed in 76 patients. QFR ≤0.80 in at least one of the three vessels was the strongest predictor of events (HR 3.14, 95%CI 1.78–5.54, p = 0.0001). This association was maintained in several sensitivity analyses. CONCLUSIONS: QFR reproducibility is acceptable, even when analysis is performed post hoc. A pathological QFR is not rare in patients without angiographic evidence of significant stenosis and is a predictor of incident events during long-term follow-up. Condensed Abstract: In a post hoc analysis of 167 patients without evidence of angiographic significant stenosis, the presence of QFR value ≤0.80 in at least one of the three coronary vessels showed to be the strongest predictor of major adverse cardiovascular and cerebrovascular events during long-term follow-up. QFR reproducibility have been shown to be acceptable among experienced operators.
Acknowledgement to Reviewers of Journal of Risk and Financial Management in 2019
Authors:   JRFM Editorial Office
Published: January 2020

The editorial team greatly appreciates the reviewers who have dedicated their considerable time and expertise to the journal’s rigorous editorial process over the past 12 months, regardless of whether the papers are finally published or not [...]
Navigating between Tea and Rubber in Xishuangbanna, China: When New Crops Fail and Old Ones Work
Authors: Slothuus Charlotte Filt , Schmidt-Vogt Dietrich , Mertz Ole
Published: January 2020

Following the massive expansion of rubber plantations in China, considerable research has been conducted on the impact of these landscape changes. The general consensus is that there have been negative impacts on the environment and positive impacts on local economies. However, since rubber prices dropped after 2011, the economic benefit to the local people is challenged and the impact on the local people and communities remains unclear. Using a mix of qualitative and quantitative methods, this longitudinal study investigates how the drop in rubber prices has affected a local community and the local people in Manlin, Xishuangbanna, China. It investigates local coping strategies and the importance of alternative income sources and shows how differentiated access to alternative lands creates increased economic inequality within the village when prices fluctuate. Three general coping strategies were identified amongst local rubber farmers: doing business as usual, changing rubber management practices, and stopping- or decreasing tapping frequency. Differences in coping strategies are linked to factors including access to alternative income sources and rubber perceptions. Moreover, households with access to tea land were found to have experienced negligible impacts of decreasing rubber prices as income from tea has increased more than income from rubber has decreased, leading to increasing intra-village economic inequality. We conclude that while this is a clear case of how income diversification is important for reducing livelihood vulnerability, it also shows that the large focus on rubber farming has created benefits in terms of improved infrastructure and connectivity that has helped expand the market for tea as well.
Novel Dental Poly (Methyl Methacrylate) Containing Phytoncide for Antifungal Effect and Inhibition of Oral Multispecies Biofilm
Authors: Lee Myung-Jin , Kim Min-Ji , Oh Sang-Hwan , Kwon Jae-Sung
Published: January 2020

Despite the many advantages of poly (methyl methacrylate) (PMMA) as a dental polymer, its antifungal and antibacterial effects remain limited. Here, phytoncide was incorporated into PMMA to inhibit fungal and biofilm accumulation without impairing the basic and biological properties of PMMA. A variable amount of phytoncide (0 wt % to 5 wt %) was incorporated into PMMA, and the basic material properties of microhardness, flexural strength and gloss were evaluated. In addition, cell viability was confirmed by MTT assay. This MTT assay measures cell viability via metabolic activity, and the color intensity of the formazan correlates viable cells. The fungal adhesion and viability on the PMMA surfaces were evaluated using Candida albicans (a pathogenic yeast). Finally, the thickness of saliva-derived biofilm was estimated. The flexural strength of PMMA decreased with increasing phytoncide contents, whereas there were no significant differences in the microhardness and gloss (p > 0.05) and the cell viability (p > 0.05) between the control and the phytoncide-incorporated PMMA samples. The amounts of adherent Candida albicans colony-forming unit (CFU) counts, and saliva-derived biofilm thickness were significantly lower in the phytoncide-incorporated PMMA compared to the control (p < 0.05). Hence, it was concluded that the incorporation of appropriate amounts of phytoncide in PMMA demonstrated antifungal effects while maintaining the properties, which could be a possible use in dentistry application such as denture base resin.
Alkali Activation of Waste Clay Bricks: Influence of The Silica Modulus, SiO2/Na2O, H2O/Na2O Molar Ratio, and Liquid/Solid Ratio
Authors: Gado R. A. , Hebda Marek , Łach Michal , Mikuła Janusz
Published: January 2020

This study was conducted to investigate the influence of various reaction conditions, namely the silica modulus SiO2/Na2O, H2O/Na2O molar ratio, and liquid/solid ratio on the geopolymerization reaction of the waste fired clay bricks (Grog). The starting raw material and the generated geopolymer specimens produced by different geopolymerization reaction conditions have been characterized using different techniques: X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and thermal analysis. Furthermore, physico–mechanical characterization has been carried out through the determination of bulk density, porosity, water absorption, and compressive strength for each sample at interval curing times of up to 28 days. The results indicated that the geopolymerization system of the waste fired clay bricks is influenced by the investigated reaction conditions at room temperature. The compressive strength of the geopolymer sample produced at optimum conditions increased significantly by up to 37.5 MPa, in comparison with 4.5 MPa for other conditions. Finally, an optimum recommendation and useful conclusions concerning the recycling and utilization of this waste material through the geopolymerization process are made for compatibility with construction applications.
Digital Image Correlation (DIC) Assessment of the Non-Linear Response of the Anterior Longitudinal Ligament of the Spine during Flexion and Extension
Authors: Ruspi Maria Luisa , Palanca Marco , Cristofolini Luca , Liebsch Christian , Villa Tomaso , Brayda-Bruno Marco , Galbusera Fabio , Wilke Hans-Joachim , La Barbera Luigi
Published: January 2020

While the non-linear behavior of spine segments has been extensively investigated in the past, the behavior of the Anterior Longitudinal Ligament (ALL) and its contribution during flexion and extension has never been studied considering the spine as a whole. The aims of the present study were to exploit Digital Image Correlation (DIC) to: (I) characterize the strain distribution on the ALL during flexion-extension, (II) compare the strain on specific regions of interest (ROI) of the ALL in front of the vertebra and of the intervertebral disc, (III) analyze the non-linear relationship between the surface strain and the imposed rotation and the resultant moment. Three specimens consisting of 6 functional spinal units (FSUs) were tested in flexion-extension. The full-field strain maps were measured on the surface of the ALL, and the most strained areas were investigated in detail. The DIC-measured strains showed different values of peak strain in correspondence with the vertebra and the disc but the average over the ROIs was of the same order of magnitude. The strain-moment curves showed a non-linear response like the moment-angle curves: in flexion the slope of the strain-moment curve was greater than in extension and with a more abrupt change of slope. To the authors’ knowledge, this is the first study addressing, by means of a full-field strain measurement, the non-linear contribution of the ALL to spine biomechanics. This study was limited to only three specimens; hence the results must be taken with caution. This information could be used in the future to build more realistic numerical models of the spine.
Structural Aspects of Decreasing the Corrosion Resistance of Zinc Coating Obtained in Baths with Al, Ni, and Pb Additives
Authors: Kania Henryk , Saternus Mariola , Kudláček Jan
Published: January 2020

The article presented the results of tests determining the synergistic effect of Al, Ni, and Pb additions on a zinc bath on the structure and corrosion resistance of coatings obtained on low silicon steel. Analyzed coatings were produced on S235JRG2 steel with Si content of 0.02 mass%. The corrosion resistance of the coatings was compared with the corrosion resistance of the coating obtained in the "pure" zinc bath. Structure at high magnifications (SEM) was determined, as well as coating thickness and chemical composition in microspheres. The corrosion resistance of the coatings was established comparatively in standard corrosion tests in neutral salt spray and a humid atmosphere containing SO2. It was found that the addition of Pb to the zinc bath reduced the corrosion resistance of the coatings. In the coating structure obtained in the Zn-AlNiPb bath, lead precipitation was observed in both the outer layer and the intermediate layer of the coating. Grain boundaries were the preferred site for lead precipitation. The presence of Pb precipitates favored conditions for the creation of additional corrosion cells, which led to a decrease in the corrosion resistance of the coatings.
Deflection of Steel Fiber Reinforced Concrete Beams Based on Waste Sand
Authors: Domski Jacek , Zakrzewski Mateusz
Published: January 2020

The article describes the selected methods of calculating the deflection of steel fiber reinforced concrete beams. Additionally, the results of the study on the deflection of steel fiber reinforced concrete beams based on waste sand are presented. This paper compares deflections measured during the four point bending test of the steel fiber reinforced, waste sand fine aggregate concrete beam with values determined in accordance with Eurocode 2, the proposal of Tan, Paramasivam, and Tan, the modified method of Alsayed, Bywalski, and Kaminski, and Amin, Foster, and Kaufmann’s method. The analysis conducted shows that the best accordance with the study and calculation results was obtained by using the modified Alsayed method.
Analyzing the Causality and Dependence between Gold Shocks and Asian Emerging Stock Markets: A Smooth Transition Copula Approach
Authors: Yamaka Woraphon , Maneejuk Paravee
Published: January 2020

This study aims to investigate the causality and dependence structure of gold shocks and Asian emerging stock markets. The positive and negative shocks of gold prices are quantified, and Granger causality-based Vector autoregressive and Copula approaches are employed to measure the causality and contagion effect, respectively, between the positive and negative gold shocks and Asian emerging stock markets’ volatilities. In addition, the nonlinear link between gold and stock markets is of concern and this motivates us to propose a Smooth Transition Dynamic Copula that allows for the structural change in time-varying dependence between gold shocks and Asian stock markets’ volatilities. Several Copula families are also considered, and the best-fit Copula model is used to explain the correlation or contagion effects. The findings of the study show that there is some significant causality between gold shocks and Asian stock markets’ volatilities in some parts of the sample period. We also observe a stronger correlation during the global financial crisis when compared to the pre- and post-crisis periods. In addition, the tail dependence is found between Indian stock and negative gold shock and between Korean stock and negative gold shock, which indicated the existence of the risk contagion effects between gold and these two stock markets.
Mathematical Model and Evaluation Function for Conflict-Free Warranted Makespan Minimization of Mixed Blocking Constraint Job-Shop Problems
Authors: Sauvey Christophe , Trabelsi Wajdi , Sauer Nathalie
Published: January 2020

In this paper, we consider a job-shop scheduling problem with mixed blocking constraints. Contrary to most previous studies, where no blocking or only one type of blocking constraint was used among successive operations, we assume that, generally, we may address several different blocking constraints in the same scheduling problem depending on the intermediate storage among machines, the characteristics of the machines, the technical constraints, and even the jobs. Our objective was to schedule a set of jobs to minimize the makespan. Thus, we propose, for the first time, a mathematical model of the job-shop problem taking into account the general case of mixed blocking constraints, and the results were obtained using Mosel Xpress software. Then, after explaining why and how groups of jobs have to be processed, a blocking constraint conflict-free warranted evaluation function is proposed and tested with the particle swarm optimization and genetic algorithm methods. The results prove that we obtained a near-optimal solution to this problem in a very short time.
Existence of Solutions for Fractional Multi-Point Boundary Value Problems on an Infinite Interval at Resonance
Authors: Zhang Wei , Liu Wenbin
Published: January 2020

This paper aims to investigate a class of fractional multi-point boundary value problems at resonance on an infinite interval. New existence results are obtained for the given problem using Mawhin’s coincidence degree theory. Moreover, two examples are given to illustrate the main results.
Marine Drugs Acting as Autophagy Modulators
Authors: Dyshlovoy Sergey A. , Honecker Friedemann
Published: January 2020

Autophagy (Ancient Greek αὐτόφαγος [autóphagos]—“self-devouring”) is defined as a regulated mechanism of the degradation of unnecessary or dysfunctional cellular components [...]
Recent Discovery of Heterocyclic Alkaloids from Marine-Derived Aspergillus Species
Authors: Xu Kuo , Yuan Xiao-Long , Li Chen , Li and Xiao-Dong
Published: January 2020

Nitrogen heterocycles have drawn considerable attention due to of their significant biological activities. The marine fungi residing in extreme environments are among the richest sources of these basic nitrogen-containing secondary metabolites. As one of the most well-known universal groups of filamentous fungi, marine-derived Aspergillus species produce a large number of structurally unique heterocyclic alkaloids. This review attempts to provide a comprehensive summary of the structural diversity and biological activities of heterocyclic alkaloids that are produced by marine-derived Aspergillus species. Herein, a total of 130 such structures that were reported from the beginning of 2014 through the end of 2018 are included, and 75 references are cited in this review, which will benefit future drug development and innovation.
Role of a 193 nm ArF Excimer Laser in Laser-Assisted Plasma-Enhanced Chemical Vapor Deposition of SiNx for Low Temperature Thin Film Encapsulation
Authors: An Kunsik , Lee Ho-Nyun , Cho Kwan Hyun , Lee Seung-Woo , Hwang David J. , Kang Kyung-Tae
Published: January 2020

In this study, silicon nitride thin films are deposited on organic polyethylene-naphthalate (PEN) substrates by laser assisted plasma enhanced chemical vapor deposition (LAPECVD) at a low temperature (150 °C) for the purpose of evaluating the encapsulation performance. A plasma generator is placed above the sample stage as conventional plasma enhanced chemical vapor deposition (PECVD) configuration, and the excimer laser beam of 193 nm wavelength illuminated in parallel to the sample surface is coupled to the reaction zone between the sample and plasma source. Major roles of the laser illumination in LAPECVD process are to compete with or complement the plasma decomposition of reactant gases. While a laser mainly decomposes ammonia molecules in the plasma, it also contributes to the photolysis of silane in the plasma state, possibly through the resulting hydrogen radicals and the excitation of intermediate disilane products. It will also be shown that the LAPECVD with coupled laser illumination of 193 nm wavelength improves the deposition rate of silicon nitride thin film, and the encapsulation performance evaluated via the measurement of water vapor transmission rate (WVTR).
Auxiliary Optomechanical Tools for 3D Cell Manipulation
Authors: Shishkin Ivan , Markovich Hen , Roichman Yael , Ginzburg Pavel
Published: January 2020

Advances in laser and optoelectronic technologies have brought the general concept of optomechanical manipulation to the level of standard biophysical tools, paving the way towards controlled experiments and measurements of tiny mechanical forces. Recent developments in direct laser writing (DLW) have enabled the realization of new types of micron-scale optomechanical tools, capable of performing designated functions. Here we further develop the concept of DLW-fabricated optomechanically-driven tools and demonstrate full-3D manipulation capabilities over biological objects. In particular, we resolved the long-standing problem of out-of-plane rotation in a pure liquid, which was demonstrated on a living cell, clamped between a pair of forks, designed for efficient manipulation with holographic optical tweezers. The demonstrated concept paves the way for the realization of flexible tools for performing on-demand functions over biological objects, such as cell tomography and surgery to name just few.
An M29 Aminopeptidase from Listeria Monocytogenes Contributes to In Vitro Bacterial Growth but not to Intracellular Infection
Authors: Zhang Xian , Guan Chiyu , Hang Yi , Liu Fengdan , Sun Jing , Yu Huifei , Gan Li , Zeng Huan , Zhu Yiran , Chen Zhongwei , Song Houhui , Cheng Changyong
Published: January 2020

Aminopeptidases that catalyze the removal of N-terminal residues from polypeptides or proteins are crucial for physiological processes. Here, we explore the biological functions of an M29 family aminopeptidase II from Listeria monocytogenes (LmAmpII). We show that LmAmpII contains a conserved catalytic motif (EEHYHD) that is essential for its enzymatic activity and LmAmpII has a substrate preference for arginine and leucine. Studies on biological roles indicate that LmAmpII is required for in vitro growth in a chemically defined medium for optimal growth of L. monocytogenes but is not required for bacterial intracellular infection in epithelial cells and macrophages, as well as cell-to-cell spreading in fibroblasts. Moreover, LmAmpII is found as dispensable for bacterial pathogenicity in mice. Taken together, we conclude that LmAmpII, an M29 family aminopeptidase, can efficiently hydrolyze a wide range of substrates and is required for in vitro bacterial growth, which lays a foundation for in-depth investigations of aminopeptidases as potential targets to defend Listeria infection.
Bioregional Alterations in Gut Microbiome Contribute to the Plasma Metabolomic Changes in Pigs Fed with Inulin
Authors: Wu Weida , Zhang Li , Xia Bing , Tang Shanlong , Liu Lei , Xie Jingjing , Zhang Hongfu
Published: January 2020

Inulin (INU) is a non-digestible carbohydrate, known for its beneficial properties in metabolic disorders. However, whether and how gut microbiota in its regulation contributes to host metabolism has yet to be investigated. We conduct this study to examine the possible associations between the gut microbiota and circulating gut microbiota–host co-metabolites induced by inulin interventions. Plasma and intestinal site samples were collected from the pigs that have consumed inulin diet for 60 days. High-throughput sequencing was adopted for microbial composition, and the GC-TOF-MS-based metabolomics were used to characterize featured plasma metabolites upon inulin intervention. Integrated multi-omics analyses were carried out to establish microbiota–host interaction. Inulin consumption decreased the total cholesterol (p = 0.04) and glucose (p = 0.03) level in serum. Greater β-diversity was observed in the cecum and colon of inulin-fed versus that of control-fed pigs (p < 0.05). No differences were observed in the ileum. In the cecum, 18 genera were altered by inulin, followed by 17 in the colon and 6 in the ileum. Inulin increased propionate, and isobutyrate concentrations but decreased the ratio of acetate to propionate in the cecum, and increased total short fatty acids, valerate, and isobutyrate concentrations in the colon. Metabolomic analysis reveals that indole-3-propionic acid (IPA) was significantly higher, and the branched-chain amino acids (BCAA), L-valine, L-isoleucine, and L-leucine are significantly lower in the inulin groups. Mantel test and integrative analysis revealed associations between plasma metabolites (e.g., IPA, BCAA, L-tryptophan) and inulin-responsive cecal microbial genera. These results indicate that the inulin has regional effects on the intestine microbiome in pigs, with the most pronounced effects occurring in the cecum. Moreover, cecum microbiota plays a pivotal role in the modulation of circulating host metabolites upon inulin intervention
Broader Geographical Distribution of Toscana Virus in the Mediterranean Region Suggests the Existence of Larger Varieties of Sand Fly Vectors
Authors: Ayhan Nazli , Prudhomme Jorian , Laroche Lison , Bañuls Anne-Laure , Charrel Remi N.
Published: January 2020

Toscana virus (TOSV) is endemic in the Mediterranean basin, where it is transmitted by sand flies. TOSV can infect humans and cause febrile illness as well as neuroinvasive infections affecting the central and peripheral nervous systems. Although TOSV is a significant human pathogen, it remains neglected and there are consequently many gaps of knowledge. Recent seroepidemiology studies and case reports showed that TOSV’s geographic distribution is much wider than was assumed a decade ago. The apparent extension of the TOSV circulation area raises the question of the sandfly species that are able to transmit the virus in natural conditions. Phlebotomus (Ph.) perniciosus and Ph. perfiliewi were historically identified as competent species. Recent results suggest that other species of sand flies could be competent for TOSV maintenance and transmission. Here we organize current knowledge in entomology, epidemiology, and virology supporting the possible existence of additional phlebotomine species such as Ph. longicuspis, Ph. sergenti, Ph. tobbi, Ph. neglectus, and Sergentomyia minuta in TOSV maintenance. We also highlight some of the knowledge gaps to be addressed in future studies.
Dietary Silk Peptide Prevents High-Fat Diet-Induced Obesity and Promotes Adipose Browning by Activating AMP-Activated Protein Kinase in Mice
Authors: Lee Kippeum , Jin Heegu , Chei Sungwoo , Lee Jeong-Yong , Oh Hyun-Ji , Lee Boo-Yong
Published: January 2020

Obesity is associated with metabolic syndrome and other chronic diseases, and is caused when the energy intake is greater than the energy expenditure. We aimed to determine the mechanism whereby acid-hydrolyzed silk peptide (SP) prevents high-fat diet-induced obesity, and whether it induces browning and fatty acid oxidation (FAO) in white adipose tissue (WAT), using in vivo and ex vivo approaches. We determined the effects of dietary SP in high-fat diet-fed obese mice. The expression of adipose tissue-specific genes was quantified by western blotting, qRT-PCR, and immunofluorescence analysis. We also investigated whether SP directly induces browning in primarily subcutaneous WAT-derived adipocytes. Our findings demonstrate that SP has a browning effect in WAT by upregulating AMP-activated Protein Kinase (AMPK) phosphorylation and uncoupling protein 1 (UCP1) expression. SP also suppresses adipogenesis and promotes FAO, implying that it may have potential as an anti-obesity drug.
The Nutritional and Micronutrient Status of Urban Schoolchildren with Moderate Anemia is Better than in a Rural Area in Kenya
Authors: Evang Esther Charlotte , Habte Tsige-Yohannes , Owino Willis Omondi , Krawinkel Michael Bernhardt
Published: January 2020

Low diet quality is a driver of general and micronutrient malnutrition in urban and rural areas. The objective was to compare malnutrition and micronutrient deficiencies linked to dietary intake among urban and rural schoolchildren from food insecure settings in Kenya. The cross-sectional study was conducted among urban and rural schoolchildren aged 7–9 years. Height and weight were measured, venous blood samples were assessed and data on dietary intake was collected. After screening out children with hemoglobin >12.2 g/dL and moderate or severe undernutrition, a total of 36 urban and 35 rural children participated. The prevalence of moderate underweight, wasting, and stunting were lower in urban than in rural children, with significant differences in median z-scores for underweight (p < 0.001) and wasting (p < 0.001). Significantly higher values for serum ferritin (p = 0.012) and zinc (p < 0.001) were found in urban children. Yet, the median adequacy ratios were higher for vitamin C (p = 0.045), iron (p = 0.003), and zinc (p = 0.003) in rural than in urban children. General nutritional, iron, and zinc status were significantly better in slightly anemic urban children than in rural ones. Improving the nutrition of schoolchildren in urban and rural settings requires different dietary approaches.
Impact of a Three-Week in-Hospital Multidisciplinary Body Weight Reduction Program on Body Composition, Muscle Performance and Fatigue in a Pediatric Obese Population with or without Metabolic Syndrome
Authors: Rigamonti Antonello Emilio , Tringali Gabriella , De Micheli Roberta , De Col Alessandra , Tamini Sofia , Saezza Antonella , Cella Silvano G. , Sartorio Alessandro
Published: January 2020

Metabolic syndrome is a combination of cardiometabolic risk factors, frequently detected in obese children and adolescents. To date, few clinical studies have evaluated the effectiveness of multidisciplinary body weight reduction programs on body mass index, body composition, muscle performance and fatigue in pediatric obese subjects suffering from metabolic syndrome, which might represent a sub-population that is more difficult to be treated and worthy of more intensive interventions than a population less metabolically complicated. The aim of the present study was to compare the impact of a three-week in-hospital multidisciplinary integrated body weight reduction program (BWRP) on body mass index (BMI), body composition (particularly, fat mass (FM) and fat-free mass (FFM)), motor control (evaluated by one-leg standing balance (OLSB) test), muscle performance (evaluated by the stair climbing test (SCT)) and fatigue (evaluated by fatigue severity scale (FSS)) in a pediatric obese population with or without metabolic syndrome. A pediatric population of 548 obese subjects without metabolic syndrome (F/M = 312/236; age range: 8–18 years; BMI: 36.3 ± 6.7 kg/m2) and 96 obese subjects with metabolic syndrome (F/M = 53/43; age range: 9–18 years; BMI: 38.3 ± 6.9 kg/m2) was recruited. The BWRP significantly reduced BMI, FM (expressed as %), SCT time and FSS score, and increased OLSB time in all subgroups of obese subjects, independent of sex and metabolic syndrome, with preservation of FFM. No significant differences in |ΔBMI|, |ΔFM|, |ΔOLSB| or |ΔSCT| times and |ΔFSS| score were found when comparing subjects (males and females) with or without metabolic syndrome, apart from obese females without metabolic syndrome, who exhibited a lower weight loss and FM (expressed as %) reduction when compared to the corresponding male counterpart. In conclusion, the beneficial effects of a three-week BWRP on BMI, body composition, muscle performance and fatigue in a pediatric obese population were not found to be different in patients with or without metabolic syndrome, thus indicating that the more metabolically compromised patient is as responsive to a short-term BWRP as the patient without metabolic syndrome. More prolonged follow-up studies are, however, necessary in order to verify whether the adherence to the multidisciplinary recommendations at home and the long-term maintenance of the positive effects in the two subgroups of patients will remain similar or not.
Validation of a Deep Learning System for the Full Automation of Bite and Meal Duration Analysis of Experimental Meal Videos
Authors: Konstantinidis Dimitrios , Dimitropoulos Kosmas , Langlet Billy , Daras Petros , Ioakimidis Ioannis
Published: January 2020

Eating behavior can have an important effect on, and be correlated with, obesity and eating disorders. Eating behavior is usually estimated through self-reporting measures, despite their limitations in reliability, based on ease of collection and analysis. A better and widely used alternative is the objective analysis of eating during meals based on human annotations of in-meal behavioral events (e.g., bites). However, this methodology is time-consuming and often affected by human error, limiting its scalability and cost-effectiveness for large-scale research. To remedy the latter, a novel “Rapid Automatic Bite Detection” (RABiD) algorithm that extracts and processes skeletal features from videos was trained in a video meal dataset (59 individuals; 85 meals; three different foods) to automatically measure meal duration and bites. In these settings, RABiD achieved near perfect agreement between algorithmic and human annotations (Cohen’s kappa κ = 0.894; F1-score: 0.948). Moreover, RABiD was used to analyze an independent eating behavior experiment (18 female participants; 45 meals; three different foods) and results showed excellent correlation between algorithmic and human annotations. The analyses revealed that, despite the changes in food (hash vs. meatballs), the total meal duration remained the same, while the number of bites were significantly reduced. Finally, a descriptive meal-progress analysis revealed that different types of food affect bite frequency, although overall bite patterns remain similar (the outcomes were the same for RABiD and manual). Subjects took bites more frequently at the beginning and the end of meals but were slower in-between. On a methodological level, RABiD offers a valid, fully automatic alternative to human meal-video annotations for the experimental analysis of human eating behavior, at a fraction of the cost and the required time, without any loss of information and data fidelity.
Effects of Capsinoids on Daily Physical Activity, Body Composition and Cold Hypersensitivity in Middle-Aged and Older Adults: A Randomized Study
Authors: Yokoyama Keiichi , Yamada Yosuke , Akamatsu Yasunori , Yoshinaka Yasuko , Yamamoto Akiko , Koizumi Tomonori , Ohyama Kana , Suzuki Katsuya , Hashimoto Masaki , Sato Hitoshi , Kimura Misaka
Published: January 2020

Sedentary/inactive lifestyle leads middle-aged and older adults to metabolic syndrome and frailty. Capsinoids from nonpungent chili pepper cultivar have been reported to reduce body fat mass, promote metabolism, and improve unidentified complaints of chills. Additionally, they have an anti-inflammation effect; therefore, we hypothesized that continuous oral ingestion of capsinoids alleviates age-related inflammation in the brain and improves the physical activity (PA) in middle-aged and older adults. In our double-blind human study, 69 participants (17 male, 52 female; mean age: 74.1 ± 7.7 years; range: 52–87 years) were administered either 9 mg of capsinoids which were extracted from pepper fruit variety CH-19 Sweet (Capsicum anuum L.) (CP group), or a placebo (PL group) daily over a 3 month period. In an animal study, PA and inflammation-related mRNA expression in the brain were examined in 5-week (young) and 53-week (old) aged mice fed a diet with or without 0.3% dihydrocapsiate, a type of capsinoids, for 12 weeks. In a human study, capsinoids intake did not increase the amount of light-to-moderate PA less than 6.0 metabolic equivalents (METs) (CP: 103.0 ± 28.2 at baseline to 108.2 ± 28.3 at 12 weeks; PL: 104.6 ± 19.8 at baseline to 115.2 ± 23.6 at 12 weeks, METs × hour/week); however, in participants exhibiting an inactive lifestyle, it showed significant increase (CP: 84.5 ± 17.2 at baseline to 99.2 ± 24.9 at 12 weeks; PL: 99.7 ± 23.3 at baseline to 103.8 ± 21.9 at 12 weeks). The energy expenditure in physical activity also improved in the inactive CP group (CP: 481.2 ± 96.3 at baseline to 562.5 ± 145.5 at 12 weeks; PL: 536.8 ± 112.2 at baseline to 598.6 ± 127.6 at 12 weeks; kcal/day). In all participants, CP showed reduced waist circumference, percent body fat, and visceral fat volume; in addition, chills were eased in subjects aged 80 years and older. The older mice fed capsinoids showed increased locomotion activity, decreased inflammation, and oxidative stress in the brain. The results suggest that the continuous oral ingestion of capsinoids gains PA through anti-inflammation effect in the brain as well as reduces fat accumulation and chills in inactive and older humans.
Evaluation of Hypoxia-Inducible Factor-1 Alpha (HIF-1α) in Equine Sarcoid: An Immunohistochemical and Biochemical Study
Authors: Martano Manuela , Altamura Gennaro , Power Karen , Restucci Brunella , Carella Francesca , Borzacchiello Giuseppe , Maiolino Paola
Published: January 2020

Background: equine sarcoids are the most frequent skin tumors in equidae worldwide. It is well known that delta bovine papillomaviruses are their causative agents. We have recently shown the presence in equine sarcoids of abnormal vessel structures, which could cause a hypoxic condition. The aim of this study was to analyze the expression of hypoxia-inducible factor-1 alpha (HIF-1α) in a subset of BPV positive equine sarcoids and explore the relationship with vascular endothelial growth factor (VEGF) expression. Results: 80% of equine sarcoids showed strong cytoplasmic staining in >60% of neoplastic fibroblasts, while 20% of samples showed a moderate cytoplasmic staining in 40–60% of neoplastic fibroblasts for HIF-1α. Results of Western blotting (WB) were consistent with immunohistochemistry (IHC). Moreover, a positive correlation between HIF-1α and VEGF expression (r = 0.60, p < 0.01) was observed. Conclusion: we have shown that HIF-1α was strongly expressed in equine sarcoid. The upregulation of HIF-1α has been described in numerous tumors and can be modulated by many proteins encoded by transforming viruses. Thus, it is also possible that BPV could have a relevant role in HIF-1α pathway regulation, contributing to the development of equine sarcoids by promoting HIF-1α/VEGF mediated tumor angiogenesis.
Acknowledgement to Reviewers of Pharmaceuticals in 2019
Authors:   Pharmaceuticals Editorial Office
Published: January 2020

The editorial team greatly appreciates the reviewers who have dedicated their considerable time and expertise to the journal’s rigorous editorial process over the past 12 months, regardless of whether the papers are finally published or not [...]
3D Printing of Drug-Loaded Thermoplastic Polyurethane Meshes: A Potential Material for Soft Tissue Reinforcement in Vaginal Surgery
Authors: Domínguez-Robles Juan , Mancinelli Caterina , Mancuso Elena , García-Romero Inmaculada , Gilmore Brendan F. , Casettari Luca , Larrañeta Eneko , Lamprou Dimitrios A.
Published: January 2020

Current strategies to treat pelvic organ prolapse (POP) or stress urinary incontinence (SUI), include the surgical implantation of vaginal meshes. Recently, there have been multiple reports of issues generated by these meshes conventionally made of poly(propylene). This material is not the ideal candidate, due to its mechanical properties leading to complications such as chronic pain and infection. In the present manuscript, we propose the use of an alternative material, thermoplastic polyurethane (TPU), loaded with an antibiotic in combination with fused deposition modelling (FDM) to prepare safer vaginal meshes. For this purpose, TPU filaments containing levofloxacin (LFX) in various concentrations (e.g., 0.25%, 0.5%, and 1%) were produced by extrusion. These filaments were used to 3D print vaginal meshes. The printed meshes were fully characterized through different tests/analyses such as fracture force studies, attenuated total reflection-Fourier transform infrared, thermal analysis, scanning electron microscopy, X-ray microcomputed tomography (μCT), release studies and microbiology testing. The results showed that LFX was uniformly distributed within the TPU matrix, regardless the concentration loaded. The mechanical properties showed that poly(propylene) (PP) is a tougher material with a lower elasticity than TPU, which seemed to be a more suitable material due to its elasticity. In addition, the printed meshes showed a significant bacteriostatic activity on both Staphylococcus aureus and Escherichia coli cultures, minimising the risk of infection after implanting them. Therefore, the incorporation of LFX to the TPU matrix can be used to prepare anti-infective vaginal meshes with enhanced mechanical properties compared with current PP vaginal meshes.
Antitumor Features of Vegetal Protein-Based Nanotherapeutics
Authors: Voci Silvia , Gagliardi Agnese , Fresta Massimo , Cosco Donato
Published: January 2020

The introduction of nanotechnology into pharmaceutical application revolutionized the administration of antitumor drugs through the modulation of their accumulation in specific organs/body compartments, a decrease in their side-effects and their controlled release from innovative systems. The use of plant-derived proteins as innovative, safe and renewable raw materials to be used for the development of polymeric nanoparticles unlocked a new scenario in the drug delivery field. In particular, the reduced size of the colloidal systems combined with the peculiar properties of non-immunogenic polymers favored the characterization and evaluation of the pharmacological activity of the novel nanoformulations. The aim of this review is to describe the physico-chemical properties of nanoparticles composed of vegetal proteins used to retain and deliver anticancer drugs, together with the most important preparation methods and the pharmacological features of these potential nanomedicines.
Ascorbate and Thiamin: Metabolic Modulators in Plant Acclimation Responses
Authors: Rosado-Souza Laise , Fernie Alisdair R. , Aarabi Fayezeh
Published: January 2020

Cell compartmentalization allows incompatible chemical reactions and localised responses to occur simultaneously, however, it also requires a complex system of communication between compartments in order to maintain the functionality of vital processes. It is clear that multiple such signals must exist, yet little is known about the identity of the key players orchestrating these interactions or about the role in the coordination of other processes. Mitochondria and chloroplasts have a considerable number of metabolites in common and are interdependent at multiple levels. Therefore, metabolites represent strong candidates as communicators between these organelles. In this context, vitamins and similar small molecules emerge as possible linkers to mediate metabolic crosstalk between compartments. This review focuses on two vitamins as potential metabolic signals within the plant cell, vitamin C (L-ascorbate) and vitamin B1 (thiamin). These two vitamins demonstrate the importance of metabolites in shaping cellular processes working as metabolic signals during acclimation processes. Inferences based on the combined studies of environment, genotype, and metabolite, in order to unravel signaling functions, are also highlighted.