bluuebunny/crossref_metadata_2025 huggingface dataset:
Methionine and Arginine Supply Alters Abundance of Amino Acid, Insulin Signaling, and Glutathione Metabolism-Related Proteins in Bovine Subcutaneous Adipose Explants Challenged with N-Acetyl-d-sphingosine
Authors: Liang Yusheng , Ma Nana , Coleman Danielle N. , Liu Fang , Li Yu , Ding Hongyan , Cardoso Fabiana F. , Parys Claudia , Cardoso Felipe C. , Loor Juan J.
Published: July 2021

The objective was to perform a proof-of-principle study to evaluate the effects of methionine (Met) and arginine (Arg) supply on protein abundance of amino acid, insulin signaling, and glutathione metabolism-related proteins in subcutaneous adipose tissue (SAT) explants under ceramide (Ce) challenge. SAT from four lactating Holstein cows was incubated with one of the following media: ideal profile of amino acid as the control (IPAA; Lys:Met 2.9:1, Lys:Arg 2:1), increased Met (incMet; Lys:Met 2.5:1), increased Arg (incArg; Lys:Arg 1:1), or incMet plus incArg (Lys:Met 2.5:1 Lys:Arg 1:1) with or without 100 μM exogenous cell-permeable Ce (N-Acetyl-d-sphingosine). Ceramide stimulation downregulated the overall abundance of phosphorylated (p) protein kinase B (AKT), p-mechanistic target of rapamycin (mTOR), and p-eukaryotic elongation factor 2 (eEF2). Without Ce stimulation, increased Met, Arg, or Met + Arg resulted in lower p-mTOR. Compared with control SAT stimulated with Ce, increased Met, Arg, or Met + Arg resulted in greater activation of mTOR (p-mTOR/total mTOR) and AKT (p-AKT/total AKT), with a more pronounced response due to Arg. The greatest protein abundance of glutathione S-transferase Mu 1 (GSTM1) was detected in response to increased Met supply during Ce stimulation. Ceramide stimulation decreased the overall protein abundance of the Na-coupled neutral amino acid transporter SLC38A1 and branched-chain alpha-ketoacid dehydrogenase kinase (BCKDK). However, compared with controls, increased Met or Arg supply attenuated the downregulation of BCKDK induced by Ce. Circulating ceramides might affect amino acid, insulin signaling, and glutathione metabolism in dairy cow adipose tissue. Further in vivo studies are needed to confirm the role of rumen-protected amino acids in regulating bovine adipose function.
Effects of Apple (Malus pomila) Pomace-Derived Pectin on the Innate Immune Responses, Expressions of Key Immune-Related Genes, Growth Performance, and Digestive Enzyme Activity of Rainbow Trout (Oncorhynchus mykiss)
Authors: Hoseinifar Seyed Hossein , Rashidian Ghasem , Ghafarifarsani Hamed , Jahazi Mohammad Amin , Soltani Mehdi , Doan Hien Van , El-Haroun Ehab , Paolucci Marina
Published: July 2021

Pectins are a group of carbohydrates found in structural parts of terrestrial plants with wide industrial and biomedical applications. This study was designed to investigate the dietary effects of apple pomace-derived pectin (APDP) in rainbow trout (Oncorhynchus mykiss). Four formulated diets were provided with different inclusion levels of APDP for 30 days: 0, 5, 10, and 20 g kg−1; referred to as control, P1, P2, and P3, respectively. In this study, 300 fish (3.56 ± 0.007 g) were randomly distributed into twelve fiberglass tanks and fed 3% of their respective body weight four times a day. At the end of the experiment, growth parameters, including weight gain, specific growth rate, and food conversion ratio (FCR) were significantly improved in P1 and P2 treatments compared to those of the other treatments. Results from proximate composition analysis showed that protein content increased, and lipid decreased in the P2 and P3 groups. Serum lysozyme, complement activity, total immunoglobulin levels, and total protein were significantly enhanced in all treatments compared to those of the control group. Gene expression results showed no significant difference in regulation of interleukin-1β (IL-1β); however, up-regulation of lysozyme, interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) was observed in both P1 and P2. Unlike lipase, the activity of protease and amylase significantly increased in fish receiving different levels of APDP compared to the control (p < 0.05). In conclusion, the present findings suggest APDA as a promising feed additive for rainbow trout.
Assessment of Meat-Type Sheep Welfare Using Animal-Based Measures
Authors: M’Hamdi Naceur , Darej Cyrine , Attia Khaoula , Guesmi Hajer , Znaïdi Ibrahim El Akram , Bouraoui Rachid , M’Hamdi Hajer , Marzouki Lamjed , Ayadi Moez
Published: July 2021

This study aimed to assess the welfare of Tunisian sheep in extensive sheep production systems using animal-based measures of ewe welfare. This study encompasses the first national survey of sheep welfare in which animal-based outcomes were tested. Animal-based welfare measures were derived from previous welfare protocols. Fifty-two Tunisian farms were studied and a number from 20 to 100 animals by flock were examinated. The whole flock was also observed to detect clinical diseases, lameness, and coughing. The human-animal relationship was selected as welfare indicators. It was evaluated through the avoidance distance test. The average avoidance distance was 10.47 ± 1.23 and 8.12 ± 0.97 m for a novel person and farmer, respectively. The global mean of body condition score (BCS) was 2.4 with 47% of ewes having a BCS of two, which may be associated with an increased risk of nutritional stress, disease, and low productivity. Ten farms had more than 7% of lambs with a low body condition score, which may be an indication of a welfare problem. The results obtained in the present study suggest that the used animal-based measures were the most reliable indicators that can be included in welfare protocols for extensive sheep production systems.
Assessing Knowledge and Perception Regarding Antimicrobial Stewardship and Antimicrobial Resistance in University Students of Pakistan: Findings and Implications
Authors: Hussain Iltaf , Yousaf Nisa , Haider Sana , Jalil Pervisha , Saleem Muhammad Usman , Imran Imran , Majeed Abdul , Rehman Anees ur , Uzair Muhammad , Rasool Muhammad Fawad , Alqahtani Faleh , Alqhtani Hussain
Published: July 2021

The irrational use of antimicrobials has enormously contributed to antimicrobial resistance (AMR) globally and especially in the developing world. To assess the knowledge and perception regarding AMR and antimicrobial stewardship (AMS), a descriptive cross-sectional study was carried out in university students enrolled in pharmacy, veterinary, and biology programs by using an online self-administered questionnaire. The Chi-square and Fisher exact tests (where applicable) were performed to assess the association of the demographics with the students’ knowledge and perception regarding AMR and AMS. A total of 496 students completed the questionnaire, among which, 85.7% of the participants were familiar with the term AMR and 79.4% of the participants correctly identified a poorly designed dosing regimen as a contributing factor towards AMR. The majority of participants (57.9%) were familiar with the term AMS and 86.5% were aware of the aim of AMS. The participants showed good knowledge regarding AMR and AMS, but to further improve student knowledge and perception of AMS and AMR, it is suggested that dedicated modules on antibiotic use and AMS should be incorporated into the curricula of these undergraduate and postgraduate programs.
Evaluating the Implementation of a Pilot Quality Improvement Program to Support Appropriate Antimicrobial Prescribing in General Practice
Authors: Biezen Ruby , Buising Kirsty , Monaghan Tim , Ball Rachael , Thursky Karin , Cheah Ron , Clark Malcolm , Manski-Nankervis Jo-Anne
Published: July 2021

Inappropriate antimicrobial prescribing contributes to increasing antimicrobial resistance. An antimicrobial stewardship (AMS) program in the form of quality improvement activities that included audit and feedback, clinical decision support and education was developed to help optimise prescribing in general practice. The aim of this study was to evaluate the implementation of this program (Guidance GP) in three general practices in Melbourne, Australia, between November 2019 and August 2020. Thirty-one general practitioners (GPs) participated in the program, with 11 GPs and three practice managers participating in follow-up focus groups and interviews to explore the acceptability and feasibility of the program. Our findings showed that the quality improvement activities were acceptable to GPs, if they accurately fit GPs’ decision-making process and workflow. It was also important that they provided clinically meaningful information in the form of audit and feedback to GPs. The time needed to coordinate the program, and costs to implement the program were some of the potential barriers identified. Facilitators of success were a “whole of practice” approach with enthusiastic GPs and practice staff, and an identified practice champion. The findings of this research will inform implementation strategies for both the Guidance GP program and AMS programs more broadly in Australian general practice, which will be critical for general practice participation and engagement.
Sperm Motility, Oxidative Status, and Mitochondrial Activity: Exploring Correlation in Different Species
Authors: Gallo Alessandra , Esposito Maria Consiglia , Tosti Elisabetta , Boni Raffaele
Published: July 2021

Sperm quality assessment is the first step for evaluating male fertility and includes the estimation of sperm concentration, motility, and morphology. Nevertheless, other parameters can be assessed providing additional information on the male reproductive potential. This study aimed to evaluate and correlate the oxidative status, mitochondrial functionality, and motility in spermatozoa of two marine invertebrate (Ciona robusta and Mytilus galloprovincialis) and one mammalian (Bos taurus) species. By combining fluorescent staining and spectrofluorometer, sperm oxidative status was evaluated through intracellular reactive oxygen species (ROS) and plasma membrane lipid peroxidation (LPO) analysis. Mitochondrial functionality was assessed through the mitochondrial membrane potential (MMP). In the three examined species, a negative correlation emerged between sperm motility vs ROS levels and LPO. Sperm motility positively correlated with MMP in bovine, whereas these parameters were not related in ascidian or even negatively related in mussel spermatozoa. MMP was negatively related to ROS and LPO levels in ascidians, only to LPO in bovine, and positively related in mussel spermatozoa. These results suggest that energy sources for sperm motility vary between species and that ROS causes a decline in sperm motility via oxidative damage of membrane lipids. Overall, this study validates the use of fluorescent probes in combination with spectrofluorometer as a simple and powerful methodology for supplementary evaluation of sperm quality shedding light on new potential quality markers and provided relevant information on sperm energetic metabolism.
A Machine Learning Algorithm for Quantitatively Diagnosing Oxidative Stress Risks in Healthy Adult Individuals Based on Health Space Methodology: A Proof-of-Concept Study Using Korean Cross-Sectional Cohort Data
Authors: Kim Youjin , Kim Yunsoo , Hwang Jiyoung , van den Broek Tim J. , Oh Bumjo , Kim Ji Yeon , Wopereis Suzan , Bouwman Jildau , Kwon Oran
Published: July 2021

Oxidative stress aggravates the progression of lifestyle-related chronic diseases. However, knowledge and practices that enable quantifying oxidative stress are still lacking. Here, we performed a proof-of-concept study to predict the oxidative stress status in a healthy population using retrospective cohort data from Boramae medical center in Korea (n = 1328). To obtain binary performance measures, we selected healthy controls versus oxidative disease cases based on the “health space” statistical methodology. We then developed a machine learning algorithm for discrimination of oxidative stress status using least absolute shrinkage and selection operator (LASSO)/elastic net regression with 10-fold cross-validation. A proposed fine-tune model included 16 features out of the full spectrum of diverse and complex data. The predictive performance was externally evaluated by generating receiver operating characteristic curves with area under the curve of 0.949 (CI 0.925 to 0.974), sensitivity of 0.923 (CI 0.879 to 0.967), and specificity of 0.855 (CI 0.795 to 0.915). Moreover, the discrimination power was confirmed by applying the proposed diagnostic model to the full dataset consisting of subjects with various degrees of oxidative stress. The results provide a feasible approach for stratifying the oxidative stress risks in the healthy population and selecting appropriate strategies for individual subjects toward implementing data-driven precision nutrition.
Performance Evaluation of a Biological Pre-Treatment Coupled with the Down-Flow Expanded Granular Bed Reactor (DEGBR) for Treatment of Poultry Slaughterhouse Wastewater
Authors: Dlamini Derrick Njabuliso , Basitere Moses , Njoya Mahomet , Ntwampe Seteno Karabo Obed , Kaskote Ephraim
Published: July 2021

Poultry slaughterhouse wastewater contains high concentrations of chemical oxygen demand (COD), total suspended solids (TSSs), fats, oil and grease (FOG), proteins and carbohydrates. It is important that the wastewater is treated to acceptable environmental discharge standards. In this study, the poultry slaughterhouse wastewater (PSW) was treated using two-stage processes consisting of a biological pre-treatment using a biodegrading agent (Eco-flushTM) coupled with a down-flow expanded granular bed reactor (DEGBR). The results showed that the biological pre-treatment was observed to be highly effective for removal of FOG, COD and TSS with a removal efficiency of 80 ± 6.3%, 38 ± 8.4% and 56 ± 7.2%, respectively. The DEGBR showed a stable performance in terms FOG, COD and, TSS removal, with average removal efficiencies of 89 ± 2.8%, 87 ± 9.5%, and 94 ± 3.7%, respectively. The overall removal rate performance of the integrated system of pre-treatment and DEGBR in terms FOG, COD and TSS, was 97 ± 0.8%, 92 ± 6.3% and 97 ± 1.2%. Furthermore, the average volatile fatty acid/alkalinity (VFA/Alkalinity) ratio of 0.2 was reported, which indicated that the DEGBR was stable throughout the operation.
Hydrothermal Carbonization of Spent Coffee Grounds
Authors: Kim Hyeok-Jin , Oh Sea-Cheon
Published: July 2021

With increasing coffee production and consumption, the amount of coffee by-product is also increasing. Therefore, there is growing worldwide interest in using these by-products as a renewable energy source. In this study, hydrothermal carbonization was conducted with subcritical water to improve the fuel characteristics of spent coffee grounds. The water content was varied, with the mass ratio between the dry sample and water set to 1:1.5 and 1:4. The reaction temperature was increased by 10 °C from 180 to 250 °C. The fuel and thermal characteristics of the reaction products were investigated through mass and energy yields, elemental, proximate, and heating value analysis. In analysis results, as the reaction temperature increased, carbon and fixed carbon content increased, and oxygen and volatile matter content decreased, resulting in an increase in calorific value. Thermogravimetric analysis, derivative thermogravimetry, and Fourier transform infrared spectroscopy were also conducted on the reaction products. To investigate their storage characteristics, chemical oxygen demand analysis was conducted. The results showed that with increasing reaction temperature, the fixed carbon content and heating value increased; also, the fuel characteristics became similar to those of coal. In addition, the reaction products became more hydrophobic as the reaction temperature increased.
Insight in the Crystallization Kinetics of AlPO4-11 Molecular Sieve Using Di-Isopropylamine as Template
Authors: Mapele Renilson O. , Silva Antonio O. S. , Souza Marcelo J. B. , Pedrosa Anne M. G. , Coriolano Ana C. F. , Fernandes Glauber J. T. , Fernandes Valter J. , Araujo Antonio S.
Published: July 2021

The hydrothermal synthesis of aluminophosphate molecular sieve type AlPO4-11 was processed from chemicals containing psueudobohemite, 85% phosphoric acid, water, and di-isopropylamine as templating agent. The crystallization of the samples was studied by taking samples in times from 2 to 74 h. The obtained white powder products were characterized by X-ray diffraction patterns (XRD), Fourier transform infrared spectroscopy (FT-IR), thermogravimetry (TG) and differential scanning calorimetry (DTG) data and pH measurement of the mother liquor. The pore volume, as determined from TG and DTG curves, was ca. 0.17 cm3g−1. The percent relative crystallinity was determined by XRD and FT-IR methods. The crystallization kinetics evidenced that the hydrothermal synthesis of AlPO4-11 exhibited in its initial phase a behavior of first order reaction with a specific velocity constant of ca. 0.25 h−1, as determined from XRD and FT-IR data. The results obtained by both X-ray diffraction and infrared spectroscopy are comparable, and FT-IR is found to be a rapid method for estimating crystallinity and structure of aluminophosphate.
Topological Navigation and Localization in Tunnels—Application to Autonomous Load-Haul-Dump Vehicles Operating in Underground Mines
Authors: Mascaró Mauricio , Parra-Tsunekawa Isao , Tampier Carlos , Ruiz-del-Solar Javier
Published: July 2021

Mobile robots are no longer used exclusively in research laboratories and indoor controlled environments, but are now also used in dynamic industrial environments, including outdoor sites. Mining is one industry where robots and autonomous vehicles are increasingly used to increase the safety of the workers, as well as to augment the productivity, efficiency, and predictability of the processes. Since autonomous vehicles navigate inside tunnels in underground mines, this kind of navigation has different precision requirements than navigating in an open environment. When driving inside tunnels, it is not relevant to have accurate self-localization, but it is necessary for autonomous vehicles to be able to move safely through the tunnel and to make appropriate decisions at its intersections and access points in the tunnel. To address these needs, a topological navigation system for mining vehicles operating in tunnels is proposed and validated in this paper. This system was specially designed to be used by Load-Haul-Dump (LHD) vehicles, also known as scoop trams, operating in underground mines. In addition, a localization system, specifically designed to be used with the topological navigation system and its associated topological map, is also proposed. The proposed topological navigation and localization systems were validated using a commercial LHD during several months at a copper sub-level stoping mine located in the Coquimbo Region in the northern part of Chile. An important aspect to be addressed when working with heavy-duty machinery, such as LHDs, is the way in which automation systems are developed and tested. For this reason, the development and testing methodology, which includes the use of simulators, scale-models of LHDs, validation, and testing using a commercial LHD in test-fields, and its final validation in a mine, are described.
Acoustic Emission Testing and Ib-Value Analysis of Ultraviolet Light-Irradiated Fiber Composites
Authors: Jung Doyun , Na Wonjin
Published: July 2021

The failure behavior of composites under ultraviolet (UV) irradiation was investigated by acoustic emission (AE) testing and Ib-value analysis. AE signals were acquired from woven glass fiber/epoxy specimens tested under tensile load. Cracks initiated earlier in UV-irradiated specimens, with a higher crack growth rate in comparison to the pristine specimen. In the UV-degraded specimen, a serrated fracture surface appeared due to surface hardening and damaged interfaces. All specimens displayed a linearly decreasing trend in Ib-values with an increasing irradiation time, reaching the same value at final failure even when the starting values were different.
A Benchmark Study of Burning Rate of Selected Thermites through an Original Gasless Theoretical Model
Authors: Brotman Sarah , Djafari Rouhani Mehdi , Charlot Samuel , Estève Alain , Rossi Carole
Published: July 2021

This paper describes a kinetic model dedicated to thermite nanopowder combustion, in which core equations are based on condensed phase mechanisms only. We explore all combinations of fuels/oxidizers, namely Al, Zr, B/CuO, Fe2O3, WO3, and Pb3O4, with 60 % of the theoretical maximum density packing, at which condensed phase mechanisms govern the reaction. Aluminothermites offer the best performances, with initiation delays in the range of a few tens of microseconds, and faster burn rates (60 cm s−1 for CuO). B and Zr based thermites are primarily limited by diffusion characteristics in their oxides that are more stringent than the common Al2O3 barrier layer. Combination of a poor thermal conductivity and efficient oxygen diffusion towards the fuel allows rapid initiation, while thermal conductivity is essential to increase the burn rate, as evidenced from iron oxide giving the fastest burn rates of all B- and Zr-based thermites (16 and 32 cm·s−1, respectively) despite poor mass transport properties in the condensed phase; almost at the level of Al/CuO (41 versus 61 cm·s−1). Finally, formulations of the effective thermal conduction coefficient are provided, from pure bulk, to nanoparticular structured material, giving light to the effects of the microstructure and its size distribution on thermite performances.
The Off-Line Simulation on Measuring through Software PC-DMIS CAD++ V4.3
Authors: Rudy Vladimír , Králik Marián , Malega Peter , Daneshjo Naqib
Published: July 2021

The article analyses and evaluates the ever-important topic of assessing geometric deviation of tolerated formations related to bases with the usage of coordinate measuring machines. The basic system for off-line simulation consists of the coordinate planes of a component’s coordinate system. At the beginning of the measurement, the coordinate system is created by the “3–2–1” alignment. Due to production deviations in real surfaces of the component, each measurement generates mutually different coordinate systems, which is well proven by the experiment on measuring with a coordinate measuring machine DEA Global Performance 12.22.10. An integral part of the article is also the quantification of geometric deviations of ideal tolerated formations related to bases, the estimate of the uncertainty of measurement arising from the placement of points in defining the base system, and the effect of such uncertainty upon the interval of satisfactory values in conformity with the STN EN ISO 14253-1 technical standard. The article also includes a proposal measure in order to ensure the reproducibility of defining the mutual position of coordinate systems.
External Shocks in the Art Markets: How Did the Portuguese, the Spanish and the Brazilian Art Markets React to COVID-19 Global Pandemic? Data Analysis and Strategies to Overcome the Crisis
Authors: Duarte Adelaide , Fialho Ana Letícia , Pérez-Ibáñez Marta
Published: July 2021

The spread of the COVID-19 pandemic worldwide, and the restrictions imposed by the social distance and the enforced confinement, are having an impact on the art markets globally. The aim of this article is to evaluate the impact of an external shock in the primary art market, using three countries as a case study: Portugal, Spain, and Brazil. These geographies have in common being at the margins in the art market’s main art hubs. It is intended to analyze how agents are responding to the new context, according to the data gathered within the gallery sector. The methods applied in the research are a combination of surveys carried out by the authors, field-based observation, along with an academic literature review, complemented by international and national reports analysis. The study’s main findings allow us to characterize the art market as a very resilient sector that energetically responded to the crisis, able to adapt and overcome challenges imposed by the new pandemic situation. Contemporary art galleries expanded digital activities, kept participating in art fairs hybrid models, continued to focus on internationalization, and pointed to the strengthening of public policies towards the sector and partnerships as key strategies to overcome the crisis.
Temporal Analysis of Urban-Suburban PET, mPET and UTCI Indices in Belgrade (Serbia)
Authors: Pecelj Milica , Matzarakis Andreas , Vujadinović Mirjam , Radovanović Milan , Vagić Nemanja , Đurić Dijana , Cvetkovic Milena
Published: July 2021

The analysis of the bioclimatic conditions is becoming increasingly relevant in climate interpretations for human needs, particularly in spatial planning, tourism, public health, sports events, bio-prognosis, etc. In this context, our study presents general temporal bioclimatic conditions in Belgrade, defined based on the PET, mPET and UTCI heat budget indices. Monthly, seasonal and annual indices were analyzed for urban and suburban weather stations based on 43 annual sets of meteorological data obtained by hourly observations at 7 h and 14 h CET. This study aims to present the distribution of PET, mPET and UTCI indices to show the pattern of each index in a mild climate location and to examine annual and seasonal differences of each index in the Belgrade urban center and suburban part of the city. The study results indicate higher biothermal stress in the urban area compared to the suburban zone and that the indices are congruent during the summer. At the same time, during the winter, they are more difficult to compare due to their peculiarities becoming more noticeable. The results obtained of all mean monthly and mean annual values of all three indices clearly indicate the difference that follows the definition of the urban heat island (UHI), particularly those from morning observation and winter season. The UTCI index shows the most significant monthly, seasonal and annual difference between urban and suburban areas for both observations. The annual difference of ΔUTCI7h amounts to 1.5 °C is the same as the annual difference of minimum temperatures (Δtmin). In contrast, the annual differences of ΔPET7h ΔmPET7h are °smaller (0.8 °C and 0.7 °C) and closer to the annual differences of maximum temperatures Δtmax amounted of 0.6 °C.
Loss of the MAF Transcription Factor in Laryngeal Squamous Cell Carcinoma
Authors: Janiszewska Joanna , Bodnar Magdalena , Paczkowska Julia , Ustaszewski Adam , Smialek Maciej J. , Szylberg Lukasz , Marszalek Andrzej , Kiwerska Katarzyna , Grenman Reidar , Szyfter Krzysztof , Wierzbicka Malgorzata , Giefing Maciej , Jarmuz-Szymczak Malgorzata
Published: July 2021

MAF is a transcription factor that may act either as a tumor suppressor or as an oncogene, depending on cell type. We have shown previously that the overexpressed miR-1290 influences MAF protein levels in LSCC (laryngeal squamous cell carcinoma) cell lines. In this study, we shed further light on the interaction between miR-1290 and MAF, as well as on cellular MAF protein localization in LSCC. We confirmed the direct interaction between miR-1290 and MAF 3′UTR by a dual-luciferase reporter assay. In addition, we used immunohistochemistry staining to analyze MAF protein distribution and observed loss of MAF nuclear expression in 58% LSCC samples, of which 10% showed complete absence of MAF, compared to nuclear and cytoplasmatic expression in 100% normal mucosa. Using TCGA data, bisulfite pyrosequencing and CNV analysis, we excluded the possibility that loss-of-function mutations, promoter region DNA methylation or CNV are responsible for MAF loss in LSCC. Finally, we identified genes involved in the regulation of apoptosis harboring the MAF binding motif in their promoter region by applied FIMO and DAVID GO analysis. Our results highlight the role of miR-1290 in suppressing MAF expression in LSCC. Furthermore, MAF loss or mislocalization in FFPE LSCC tumor samples might suggest that MAF acts as a LSCC tumor suppressor by regulating apoptosis.
Anti-Proliferative and Anti-Migratory Activities of Hispidulin on Human Melanoma A2058 Cells
Authors: Chang Chi-Jen , Hung Yen-Ling , Chen Ting-Chen , Li Hsin-Ju , Lo Yuan-Hsin , Wu Nan-Lin , Chang Der-Chen , Hung Chi-Feng
Published: July 2021

Melanoma represents less than 5% of skin cancers, but is the most lethal, mainly because of its high-metastatic potential and resistance to various therapies. Therefore, it is important to develop effective treatments, especially chemotherapeutic drugs with cytotoxicity, anti-metastaticity, and few side effects. One such natural product is hispidulin, a flavone distributed in plants of the Asteraceae. Previous studies have demonstrated that hispidulin has various pharmacological benefits, such as anti-tumor, anti-inflammation, and anti-allergic effects. This study aims to explore the effects of hispidulin against melanoma in vitro and in vivo. Results revealed that hispidulin selectively decreased the cell viability of A2058 cells in a dose- and time-dependent manner. Hispidulin induced cells accumulated in the sub-G1 phase via activating caspase 8 and 9, increased cleaved caspase 3, and cleaved PARP expression. Hispidulin was able to decrease AKT and ERK phosphorylation, which facilitated cell growth and survival. Moreover, hispidulin promoted reactive oxygen species generation in cells and suppressed cell migration through downregulated matrix metalloproteinase-2 expression. Hispidulin significantly inhibited tumor growth in a xenograft model. Based on these results, hispidulin produces its anti-melanoma effects by inducing cancer cell apoptosis and reducing its migration. Therefore, we suggest hispidulin as a potent therapeutic candidate for melanoma treatment.
Systematic Structure-Based Search for Ochratoxin-Degrading Enzymes in Proteomes from Filamentous Fungi
Authors: Leitão Ana Lúcia , Enguita Francisco J.
Published: July 2021

(1) Background: ochratoxins are mycotoxins produced by filamentous fungi with important implications in the food manufacturing industry due to their toxicity. Decontamination by specific ochratoxin-degrading enzymes has become an interesting alternative for the treatment of contaminated food commodities. (2) Methods: using a structure-based approach based on homology modeling, blind molecular docking of substrates and characterization of low-frequency protein motions, we performed a proteome mining in filamentous fungi to characterize new enzymes with potential ochratoxinase activity. (3) Results: the proteome mining results demonstrated the ubiquitous presence of fungal binuclear zinc-dependent amido-hydrolases with a high degree of structural homology to the already characterized ochratoxinase from Aspergillus niger. Ochratoxinase-like enzymes from ochratoxin-producing fungi showed more favorable substrate-binding pockets to accommodate ochratoxins A and B. (4) Conclusions: filamentous fungi are an interesting and rich source of hydrolases potentially capable of degrading ochratoxins, and could be used for the detoxification of diverse food commodities.
In Silico Analysis of the Enzymes Involved in Haloarchaeal Denitrification
Authors: Bernabeu Eric , Miralles-Robledillo Jose María , Giani Micaela , Valdés Elena , Martínez-Espinosa Rosa María , Pire Carmen
Published: July 2021

During the last century, anthropogenic activities such as fertilization have led to an increase in pollution in many ecosystems by nitrogen compounds. Consequently, researchers aim to reduce nitrogen pollutants following different strategies. Some haloarchaea, owing to their denitrifier metabolism, have been proposed as good model organisms for the removal of not only nitrate, nitrite, and ammonium, but also (per)chlorates and bromate in brines and saline wastewater. Bacterial denitrification has been extensively described at the physiological, biochemical, and genetic levels. However, their haloarchaea counterparts remain poorly described. In previous work the model structure of nitric oxide reductase was analysed. In this study, a bioinformatic analysis of the sequences and the structural models of the nitrate, nitrite and nitrous oxide reductases has been described for the first time in the haloarchaeon model Haloferax mediterranei. The main residues involved in the catalytic mechanism and in the coordination of the metal centres have been explored to shed light on their structural characterization and classification. These results set the basis for understanding the molecular mechanism for haloarchaeal denitrification, necessary for the use and optimization of these microorganisms in bioremediation of saline environments among other potential applications including bioremediation of industrial waters.
Bisphenol a Exposure and Kidney Diseases: Systematic Review, Meta-Analysis, and NHANES 03–16 Study
Authors: Moreno-Gómez-Toledano Rafael , Arenas María I. , Vélez-Vélez Esperanza , Coll Elisabeth , Quiroga Borja , Bover Jordi , Bosch Ricardo J.
Published: July 2021

Bisphenol A (BPA) is a compound that is especially widespread in most commonly used objects due to its multiple uses in the plastic industry. However, several data support the need to restrict its use. In recent years, new implications of BPA on the renal system have been discovered, which denotes the need to expand studies in patients. To this end, a systematic review and a meta-analysis was performed to explore existing literature that examines the BPA-kidney disease paradigm and to determine what and how future studies will need to be carried out. Our systematic review revealed that only few relevant publications have focused on the problem. However, the subsequent meta-analysis revealed that high blood concentrations of BPA could be a factor in developing kidney disease, at least in people with previous pathologies such as diabetes or hypertension. Furthermore, BPA could also represent a risk factor in healthy people whose urinary excretion is higher. Finally, the data analyzed from the NHANES 03-16 cohort provided new evidence on the possible involvement of BPA in kidney disease. Therefore, our results underline the need to carry out a thorough and methodologically homogeneous study, delving into the relationship between urinary and blood BPA, glomerular filtration rate, and urine albumin-to-creatinine ratio, preferably in population groups at risk, and subsequently in the general population, to solve this relevant conundrum with critical potential implications in Public Health.
Antibody Protection against Long-Term Memory Loss Induced by Monomeric C-Reactive Protein in a Mouse Model of Dementia
Authors: García-Lara Elisa , Aguirre Samuel , Clotet Núria , Sawkulycz Xenia , Bartra Clara , Almenara-Fuentes Lidia , Suñol Cristina , Corpas Rubén , Olah Peter , Tripon Florin , Crauciuc Andrei , Slevin Mark , Sanfeliu Coral
Published: July 2021

Monomeric C-reactive protein (mCRP), the activated isoform of CRP, induces tissue damage in a range of inflammatory pathologies. Its detection in infarcted human brain tissue and its experimentally proven ability to promote dementia with Alzheimer’s disease (AD) traits at 4 weeks after intrahippocampal injection in mice have suggested that it may contribute to the development of AD after cerebrovascular injury. Here, we showed that a single hippocampal administration of mCRP in mice induced memory loss, lasting at least 6 months, along with neurodegenerative changes detected by increased levels of hyperphosphorylated tau protein and a decrease of the neuroplasticity marker Egr1. Furthermore, co-treatment with the monoclonal antibody 8C10 specific for mCRP showed that long-term memory loss and tau pathology were entirely avoided by early blockade of mCRP. Notably, 8C10 mitigated Egr1 decrease in the mouse hippocampus. 8C10 also protected against mCRP-induced inflammatory pathways in a microglial cell line, as shown by the prevention of increased generation of nitric oxide. Additional in vivo and in vitro neuroprotective testing with the anti-inflammatory agent TPPU, an inhibitor of the soluble epoxide hydrolase enzyme, confirmed the predominant involvement of neuroinflammatory processes in the dementia induced by mCRP. Therefore, locally deposited mCRP in the infarcted brain may be a novel biomarker for AD prognosis, and its antibody blockade opens up therapeutic opportunities for reducing post-stroke AD risk.
Pulmonary MicroRNA Changes Alter Angiogenesis in Chronic Obstructive Pulmonary Disease and Lung Cancer
Authors: Green Clara E. , Clarke Joseph , Bicknell Roy , Turner Alice M.
Published: July 2021

The pulmonary endothelium is dysfunctional in chronic obstructive pulmonary disease (COPD), a known risk factor for lung cancer. The pulmonary endothelium is altered in emphysema, which is disproportionately affected by cancers. Gene and microRNA expression differs between COPD and non-COPD lung. We hypothesised that the alteration in microRNA expression in the pulmonary endothelium contributes to its dysfunction. A total of 28 patients undergoing pulmonary resection were recruited and endothelial cells were isolated from healthy lung and tumour. MicroRNA expression was compared between COPD and non-COPD patients. Positive findings were confirmed by quantitative polymerase chain reaction (qPCR). Assays assessing angiogenesis and cellular migration were conducted in Human Umbilical Vein Endothelial Cells (n = 3–4) transfected with microRNA mimics and compared to cells transfected with negative control RNA. Expression of miR-181b-3p, miR-429 and miR-23c (all p < 0.05) was increased in COPD. Over-expression of miR-181b-3p was associated with reduced endothelial sprouting (p < 0.05). miR-429 was overexpressed in lung cancer as well and exhibited a reduction in tubular formation. MicroRNA-driven changes in the pulmonary endothelium thus represent a novel mechanism driving emphysema. These processes warrant further study to determine if they may be therapeutic targets in COPD and lung cancer.
Development of the Sensing Platform for Protein Tyrosine Kinase Activity
Authors: Wei Lan-Yi , Lin Wei , Leo Bey-Fen , Kiew Lik-Voon , Chang Chia-Ching , Yuan Chiun-Jye
Published: July 2021

A miniature tyrosinase-based electrochemical sensing platform for label-free detection of protein tyrosine kinase activity was developed in this study. The developed miniature sensing platform can detect the substrate peptides for tyrosine kinases, such as c-Src, Hck and Her2, in a low sample volume (1–2 μL). The developed sensing platform exhibited a high reproducibility for repetitive measurement with an RSD (relative standard deviation) of 6.6%. The developed sensing platform can detect the Hck and Her2 in a linear range of 1–200 U/mL with the detection limit of 1 U/mL. The sensing platform was also effective in assessing the specificity and efficacies of the inhibitors for protein tyrosine kinases. This is demonstrated by the detection of significant inhibition of Hck (~88.1%, but not Her2) by the Src inhibitor 1, an inhibitor for Src family kinases, as well as the significant inhibition of Her2 (~91%, but not Hck) by CP-724714 through the platform. These results suggest the potential of the developed miniature sensing platform as an effective tool for detecting different protein tyrosine kinase activity and for accessing the inhibitory effect of various inhibitors to these kinases.
Sensing Hydration of Biomimetic Cell Membranes
Authors: Chattopadhyay Madhurima , Orlikowska Hanna , Krok Emilia , Piatkowski Lukasz
Published: July 2021

Biological membranes play a vital role in cell functioning, providing structural integrity, controlling signal transduction, and controlling the transport of various chemical species. Owing to the complex nature of biomembranes, the self-assembly of lipids in aqueous media has been utilized to develop model systems mimicking the lipid bilayer structure, paving the way to elucidate the mechanisms underlying various biological processes, as well as to develop a number of biomedical and technical applications. The hydration properties of lipid bilayers are crucial for their activity in various cellular processes. Of particular interest is the local membrane dehydration, which occurs in membrane fusion events, including neurotransmission, fertilization, and viral entry. The lack of universal technique to evaluate the local hydration state of the membrane components hampers understanding of the molecular-level mechanisms of these processes. Here, we present a new approach to quantify the hydration state of lipid bilayers. It takes advantage of the change in the lateral diffusion of lipids that depends on the number of water molecules hydrating them. Using fluorescence recovery after photobleaching technique, we applied this approach to planar single and multicomponent supported lipid bilayers. The method enables the determination of the hydration level of a biomimetic membrane down to a few water molecules per lipid.
Risk Classification for Overall Survival by the Neutrophil–Lymphocyte Ratio and the Number of Metastatic Sites in Patients Treated with Pembrolizumab—A Multicenter Collaborative Study in Japan
Authors: Uchimoto Taizo , Komura Kazumasa , Fukuokaya Wataru , Kimura Takahiro , Takahashi Kazuhiro , Yano Yusuke , Nishimura Kazuki , Nakamori Keita , Fujiwara Yuya , Matsunaga Tomohisa , Tsutsumi Takeshi , Tsujino Takuya , Maenosono Ryoichi , Yoshikawa Yuki , Taniguchi Kohei , Tanaka Tomohito , Uehara Hirofumi , Hirano Hajime , Nomi Hayahito , Takahara Kiyoshi , Inamoto Teruo , Egawa Shin , Azuma Haruhito
Published: July 2021

Pembrolizumab has emerged as the new standard of care in patients with platinum-refractory metastatic urothelial carcinoma (mUC), whereas the optimal risk stratification to predict survival outcomes is still controversial. We examined a risk model for overall survival (OS) in mUC treated with pembrolizumab using our multi-institutional dataset (212 patients). The median age was 72 years old. Median OS from the initiation of pembrolizumab treatment was 11.7 months. The objective response rate (ORR) was 26.4%. On multivariate analysis, multiple metastatic sites and an NLR > 3.50 at the initiation of pembrolizumab treatment were identified as independent predictors for OS. We next developed a risk model using those two predictors. Patients without any factors were assigned to the favorable-risk group (26.5%). Patients with either factor and both factors were assigned to the intermediate-risk group (44.3%), and poor-risk group (29.2%), respectively. Kaplan–Meier curves showed clear discrimination of OS among the risk groups (p < 0.001). The ORR in each group was 35.7% in the favorable-risk group, 27.7% in the intermediate-risk group, and 17.7% in the poor-risk group. Given that the model can be concisely determined at the initiation of pembrolizumab treatment, physicians may be encouraged to consider the risk group for daily practice.
Intensity Modulated Proton Beam Therapy versus Volumetric Modulated Arc Therapy for Patients with Nasopharyngeal Cancer: A Propensity Score-Matched Study
Authors: Chou Yung-Chih , Fan Kang-Hsing , Lin Chien-Yu , Hung Tsung-Min , Huang Bing-Shen , Chang Kai-Ping , Kang Chung-Jan , Huang Shiang-Fu , Chang Po-Hung , Hsu Cheng-Lung , Wang Hung-Ming , Hsieh Jason Chia-Hsun , Cheng Ann-Joy , Chang Joseph Tung-Chieh
Published: July 2021

(1) Background: We compared the outcomes of patients with nasopharyngeal carcinoma treated with IMPT and VMAT. (2) Methods: We performed a retrospective propensity score matching analysis (1:1) of patients treated with IMPT (years: 2016–2018) and VMAT (2014–2018). Survival was estimated using the Kaplan–Meier method. Multivariate Cox proportional hazards regression analysis was used to identify the independent predictors of survival. Binary toxicity endpoint analyses were performed using a Cox model and logistic regression. (3) Results: Eighty patients who received IMPT and VMAT were included. The median follow-up time was 24.1 months in the IMPT group. Progression-free survival (PFS) and overall survival (OS) were not statistically different between the two groups but potentially better in IMPT group. In multivariate analysis, advanced N-stage and body weight loss (BWL; >7%) during radiotherapy were associated with decreased PFS. The IMPT group had significantly less requirement for nasogastric (NG) tube placement and BWL during treatment. The mean oral cavity dose was the only predictive factor in stepwise regression analysis, and IMPT required a significantly lower mean dose. However, IMPT increased the grade 3 radiation dermatitis. (4) Conclusions: IMPT is associated with reduced rates of NG tube insertion and BWL through reducing oral mean dose, potentially producing better oncologic outcomes.
TBC1D9: An Important Modulator of Tumorigenesis in Breast Cancer
Authors: Kothari Charu , Clemenceau Alisson , Ouellette Geneviève , Ennour-Idrissi Kaoutar , Michaud Annick , C.-Gaudreault René , Diorio Caroline , Durocher Francine
Published: July 2021

Triple-negative breast cancer (TNBC) is a major concern among the different subtypes of breast cancer (BC) due to the lack of effective treatment. In a previous study by our group aimed at understanding the difference between TNBC and non-TNBC tumors, we identified the gene TBC1 domain family member 9 (TBC1D9), the expression of which was lower in TNBC as compared to non-TNBC tumors. In the present study, analysis of TBC1D9 expression in TNBC (n = 58) and non-TNBC (n = 25) patient tumor samples validated that TBC1D9 expression can differentiate TNBC (low) from non-TNBC (high) samples and that expression of TBC1D9 was inversely correlated with grade and proliferative index. Moreover, we found that downregulation of the TBC1D9 gene decreases the proliferation marginally in non-TNBC and was associated with increased migratory and tumorigenic potential in both TNBC and luminal BC cell lines. This increase was mediated by the upregulation of ARL8A, ARL8B, PLK1, HIF1α, STAT3, and SPP1 expression in TBC1D9 knockdown cells. Our results suggest that TBC1D9 expression might limit tumor aggressiveness and that it has a differential expression in TNBC vs. non-TNBC tumors.
Therapeutic Application of Brain-Specific Angiogenesis Inhibitor 1 for Cancer Therapy
Authors: Nair Mitra , Bolyard Chelsea , Lee Tae Jin , Kaur Balveen , Yoo Ji Young
Published: July 2021

Brain-specific angiogenesis inhibitor 1 (BAI1/ADGRB1) is an adhesion G protein-coupled receptor that has been found to play key roles in phagocytosis, inflammation, synaptogenesis, the inhibition of angiogenesis, and myoblast fusion. As the name suggests, it is primarily expressed in the brain, with a high expression in the normal adult and developing brain. Additionally, its expression is reduced in brain cancers, such as glioblastoma (GBM) and peripheral cancers, suggesting that BAI1 is a tumor suppressor gene. Several investigators have demonstrated that the restoration of BAI1 expression in cancer cells results in reduced tumor growth and angiogenesis. Its expression has also been shown to be inversely correlated with tumor progression, neovascularization, and peri-tumoral brain edema. One method of restoring BAI1 expression is by using oncolytic virus (OV) therapy, a strategy which has been tested in various tumor models. Oncolytic herpes simplex viruses engineered to express the secreted fragment of BAI1, called Vasculostatin (Vstat120), have shown potent anti-tumor and anti-angiogenic effects in multiple tumor models. Combining Vstat120-expressing oHSVs with other chemotherapeutic agents has also shown to increase the overall anti-tumor efficacy in both in vitro and in vivo models. In the current review, we describe the structure and function of BAI1 and summarize its application in the context of cancer treatment.