bluuebunny/crossref_metadata_2025 huggingface dataset:
Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics
Authors: Williams Holden C. , Piron Margaret A. , Nation Grant K. , Walsh Adeline E. , Young Lyndsay E. A. , Sun Ramon C. , Johnson Lance A.
Published: December 2020

Stable isotope-resolved metabolomics (SIRM) is a powerful tool for understanding disease. Advances in SIRM techniques have improved isotopic delivery and expanded the workflow from exclusively in vitro applications to in vivo methodologies to study systemic metabolism. Here, we report a simple, minimally-invasive and cost-effective method of tracer delivery to study SIRM in vivo in laboratory mice. Following a brief fasting period, we orally administered a solution of [U-13C] glucose through a blunt gavage needle without anesthesia, at a physiological dose commonly used for glucose tolerance tests (2 g/kg bodyweight). We defined isotopic enrichment in plasma and tissue at 15, 30, 120, and 240 min post-gavage. 13C-labeled glucose peaked in plasma around 15 min post-gavage, followed by period of metabolic decay and clearance until 4 h. We demonstrate robust enrichment of a variety of central carbon metabolites in the plasma, brain and liver of C57/BL6 mice, including amino acids, neurotransmitters, and glycolytic and tricarboxylic acid (TCA) cycle intermediates. We then applied this method to study in vivo metabolism in two distinct mouse models of diseases known to involve dysregulation of glucose metabolism: Alzheimer’s disease and type II diabetes. By delivering [U-13C] glucose via oral gavage to the 5XFAD Alzheimer’s disease model and the Lepob/ob type II diabetes model, we were able to resolve significant differences in multiple central carbon pathways in both model systems, thus providing evidence of the utility of this method to study diseases with metabolic components. Together, these data clearly demonstrate the efficacy and efficiency of an oral gavage delivery method, and present a clear time course for 13C enrichment in plasma, liver and brain of mice following oral gavage of [U-13C] glucose—data we hope will aid other researchers in their own 13C-glucose metabolomics study design.
Prediction of Autoimmune Diseases by Targeted Metabolomic Assay of Urinary Organic Acids
Authors: Tsoukalas Dimitris , Fragoulakis Vassileios , Papakonstantinou Evangelos , Antonaki Maria , Vozikis Athanassios , Tsatsakis Aristidis , Buga Ana Maria , Mitroi Mihaela , Calina Daniela
Published: December 2020

Autoimmune diseases (ADs) are chronic disorders characterized by the loss of self-tolerance, and although being heterogeneous, they share common pathogenic mechanisms. Self-antigens and inflammation markers are established diagnostic tools; however, the metabolic imbalances that underlie ADs are poorly described. The study aimed to employ metabolomics for the detection of disease-related changes in autoimmune diseases that could have predictive value. Quantitative analysis of 28 urine organic acids was performed using Gas Chromatography-Mass Spectrometry in a group of 392 participants. Autoimmune thyroiditis, inflammatory bowel disease, psoriasis and rheumatoid arthritis were the most prevalent autoimmune diseases of the study. Statistically significant differences were observed in the tricarboxylate cycle metabolites, succinate, methylcitrate and malate, the pyroglutamate and 2-hydroxybutyrate from the glutathione cycle and the metabolites methylmalonate, 4-hydroxyphenylpyruvate, 2-hydroxyglutarate and 2-hydroxyisobutyrate between the AD group and the control. Artificial neural networks and Binary logistic regression resulted in the highest predictive accuracy scores (66.7% and 74.9%, respectively), while Methylmalonate, 2-Hydroxyglutarate and 2-hydroxybutyrate were proposed as potential biomarkers for autoimmune diseases. Urine organic acid levels related to the mechanisms of energy production and detoxification were associated with the presence of autoimmune diseases and could be an adjunct tool for early diagnosis and prediction.
Effects of Dietary Defatted Meat Species on Metabolomic Profiles of Murine Liver, Gastrocnemius Muscle, and Cecal Content
Authors: Nakata Rise , Sato Mikako , Tomonaga Shozo
Published: December 2020

In both humans and animals, meat not only constitutes one of the sources of protein, but also includes various water-soluble bioactive substances such as imidazole peptides (carnosine and anserine) and taurine. Previous studies demonstrated that dietary meat species could differently affect physiological functions; however, the mechanisms of this remain unclear. To explore the physiological effects of dietary defatted meat species, especially on metabolism, we investigated their influence on the metabolomic profiles of the liver, gastrocnemius muscle, and cecal content in mice. Casein (control) or a defatted meat species (beef leg, pork leg, chicken leg, or chicken breast) was supplied as the major protein source in the diet for four weeks, and metabolism-related molecules were measured by gas chromatography–mass spectrometry. We found that various metabolite levels in tissues and cecal content differed according to the types of dietary protein consumed. Specifically, differences in carnosine, 1,5-anhydro-glucitol, inositol, butyric acid, and propionic acid were clearly observed. Among them, the highest carnosine intake by dietary pork leg was clearly related to the highest carnosine level in the liver. In addition, taurine intake was suggested to be linked to some metabolic pathways including taurine and hypotaurine metabolism in cecal content. These results provide additional knowledge of the effects of different dietary meat species on human and animal health.
A Framework for Evaluating Variation in (U-Th)/He Datasets
Authors: Sousa Francis J. , Farley Kenneth A.
Published: December 2020

This paper presents a framework for evaluating variation in (U-Th)/He datasets. The framework is objective, repeatable, and based on compatibility of thermal histories derived from individual (U-Th)/He dates. The structure of this new method includes three fundamental steps. First, the allowable thermal history of each individual grain is quantitatively constrained with a model. Second, the thermal histories of all grains from a sample are visualized on the same axes. Third, the compatibility of the allowable thermal histories of each individual grain is evaluated. This allows a user to assess whether variation among single grain dates can plausibly be explained (referred to here as legitimate) or not (illegitimate). Additionally, this methodology allows for accurate representation of the impact that illegitimate variation has on the thermal history constraints of a sample. We demonstrate the application of this new framework using a variety of examples from the literature, as well as with synthetic data. Modeling presented here is executed using the modeling software QTQt (version 5.6.0) and the He diffusion kinetics based on the radiation damage accumulation and annealing model, but the framework is designed to be easily adaptable to any modeling software and diffusion parameters.
Phase Behavior and Polymorphism of Saturated and Unsaturated Phytosterol Esters
Authors: Daels Eva , Foubert Imogen , Guo Zheng , Thielemans Wim , Goderis Bart
Published: December 2020

This study investigated how the physicochemical characteristics of phytosterol esters are influenced by the chain length and degree of unsaturation of the fatty acid ester moiety. Saturated and unsaturated phytosterol esters (PEs) were synthesized by the esterification of different types of fatty acids (stearic, palmitic, lauric, oleic, and linoleic acid) to β-sitosterol. The non-isothermal crystallization and melting behavior of the pure PEs were analyzed. It was proven by X-ray diffraction that saturated β-sitosteryl esters and β-sitosteryl oleate formed a bilayer crystal structure. The lamellar spacings of the bilayer structure decreased with decreasing fatty acid chain length and with an increasing degree in unsaturation. The degree of unsaturation of the fatty acid chain of the β-sitosteryl esters also influenced the type of subcell packing of the fatty acid moieties in the bilayer structure, whether or not a metastable or stable liquid crystalline phase was formed during cooling. Furthermore, it was found that the melting temperature and enthalpy of the β-sitosteryl esters increased with an increasing fatty acid chain length while they decreased with an increasing degree of unsaturation. The microscopic analyses demonstrated that β-sitosteryl oleate formed much smaller spherulites than their saturated β-sitosteryl analogues.
Characterization of Bioactive Compounds of Opuntia ficus-indica (L.) Mill. Seeds from Spanish Cultivars
Authors: Kolniak-Ostek Joanna , Kita Agnieszka , Miedzianka Joanna , Andreu-Coll Lucia , Legua Pilar , Hernandez Francisca
Published: December 2020

Opuntia ficus-indica (L.) Mill. is the Cactaceae plant with the greatest economic relevance in the world. It can be used for medicinal purposes, animal nutrition, production of biofuels and phytoremediation of soils. Due to its high content of bioactive compounds, the prickly pear has antioxidant, antimicrobial and anticancer properties. The aim of this study was to determine the polyphenolic, fatty acid and amino acid profile and characterize the antioxidant capacity of seeds of seven Spanish prickly pear cultivars. A total of 21 metabolites, mainly phenolic acids and flavonols, were identified using ultraperformance liquid chromatography photodiode detector quadrupole/time-of-flight mass spectrometry (UPLC-PDA-Q/TOF-MS). Significant differences were found in the phenolic concentrations of the investigated varieties. The highest amount of phenolic compounds (266.67 mg/kg dry matter) were found in the “Nopal espinoso” variety, while the “Fresa” variety was characterized by the lowest content (34.07 mg/kg DM) of these compounds. In vitro antioxidant capacity was positively correlated with the amount of polyphenols. The amino acid composition of protein contained in prickly pear seeds was influenced by the variety. Glutamic acid was the predominant amino acid followed by arginine, aspartic acid and leucine, independent of prickly pear variety. Overall, 13 different fatty acids were identified and assessed in prickly pear seeds. The dominant fatty acid was linoleic acid, with content varying between 57.72% “Nopal ovalado” and 63.11% “Nopal espinoso”.
Biosourced All-Acrylic ABA Block Copolymers with Lactic Acid-Based Soft Phase
Authors: Bensabeh Nabil , Jiménez-Alesanco Ana , Liblikas Ilme , Ronda Juan C. , Cádiz Virginia , Galià Marina , Vares Lauri , Abián Olga , Lligadas Gerard
Published: December 2020

Lactic acid is one of the key biobased chemical building blocks, given its readily availability from sugars through fermentation and facile conversion into a range of important chemical intermediates and polymers. Herein, well-defined rubbery polymers derived from butyl lactate solvent were successfully prepared by reversible addition–fragmentation chain transfer (RAFT) polymerization of the corresponding monomeric acrylic derivative. Good control over molecular weight and molecular weight distribution was achieved in bulk using either monofunctional or bifunctional trithiocarbonate-type chain transfer agents. Subsequently, poly(butyl lactate acrylate), with a relative low Tg (−20 °C), good thermal stability (5% wt. loss at 340 °C) and low toxicity was evaluated as a sustainable middle block in all-acrylic ABA copolymers using isosorbide and vanillin-derived glassy polyacrylates as representative end blocks. Thermal, morphological and mechanical properties of copolymers containing hard segment contents of <20 wt% were evaluated to demonstrate the suitability of rubbery poly(alkyl lactate) building blocks for developing functional sustainable materials. Noteworthy, 180° peel adhesion measurements showed that the synthesized biosourced all-acrylic ABA copolymers possess competitive performance when compared with commercial pressure-sensitive tapes.
Effect of DODAB Nano-Sized Cationic Bilayer Fragments against Leishmania amazonensis
Authors: Ferreira Thalita C. S. , Sauter Ismael P. , Borda-Samper Lina , Bentivoglio Enyd , DaMata Jarina P. , Taniwaki Noemi N. , Orrego Patrício R. , Araya Jorge E. , Lincopan Nilton , Cortez Mauro
Published: December 2020

The dioctadecyldimethylammonium bromide (DODAB) is a double-chained cationic lipid with potent bactericide and fungistatic activities; however, its toxicity on protozoan parasites is still unknown. Here, we show the antileishmanial activity of DODAB nano-sized cationic bilayer fragments on stationary-phase promastigotes and amastigotes of Leishmania amazonensis, the causative agent of cutaneous leishmaniasis. Upon treatment with DODAB, we analyzed the parasite surface zeta-potential, parasite viability, cellular structural modifications, and intracellular proliferation. The DODAB cytotoxic effect was dose-dependent, with a median effective concentration (EC50) of 25 µM for both life-cycle stages, comparable to the reported data for bacteria and fungi. The treatment with DODAB changed the membrane zeta-potential from negative to positive, compromised the parasite’s morphology, affected the cell size regulation, caused a loss of intracellular organelles, and probably dysregulated the plasma membrane permeability without membrane disruption. Moreover, the parasites that survived after treatment induced small parasitophorous vacuoles and failed to proliferate inside macrophages. In conclusion, DODAB displayed antileishmanial activity, and it remains to be elucidated how DODAB acts on the protozoan membrane. Understanding this mechanism can provide insights into the development of new parasite-control strategies.
Fe(III) Complexes Based on Mono- and Bis-pyrazolyl-s-triazine Ligands: Synthesis, Molecular Structure, Hirshfeld, and Antimicrobial Evaluations
Authors: Soliman Saied M. , Al-Rasheed Hessa H. , Albering Jörg H. , El-Faham Ayman
Published: December 2020

The self-assembly of iron(III) chloride with three pyrazolyl-s-triazine ligands, namely 2,4-bis(3,5-dimethyl-1H-pyrazol-1-yl)-6-(piperidin-1-yl)-1,3,5-triazine (PipBPT), 4-(4,6-bis(3,5-dimethyl-1H-pyrazol-1-yl)-1,3,5-triazin-2-yl)morpholine (MorphBPT), and 4,4’-(6-(3,5-dimethyl-1H-pyrazol-1-yl)-1,3,5-triazine-2,4-diyl)dimorpholine (bisMorphPT) afforded [Fe(PipBPT)Cl2][FeCl4] (1), [Fe(MorphBPT)Cl2][FeCl4] (2), and [H(bisMorphPT)][FeCl4]. bisMorphPT.2H2O (3), respectively, in good yield. In complexes 1 and 2, the Fe(III) is pentacoordinated with three Fe-N interactions from the pincer ligand and two coordinated chloride anions in the inner sphere, and FeCl4¯ in the outer sphere. Complex 3 is comprised of one protonated ligand as cationic part, one FeCl4¯ anion, and one neutral bisMorphPT molecule in addition to two crystallized water molecules. Analysis of molecular packing using Hirshfeld calculations indicated that H…H and Cl…H are the most important in the molecular packing. They comprised 40.1% and 37.4%, respectively in 1 and 32.4% and 37.8%, respectively in 2. Complex 1 exhibited the most bioactivity against the tested microbes while 3 had the lowest bioactivity. The bisMorphPT and MorphBPT were inactive towards the tested microbes while PipBPT was active. As a whole, the Fe(III) complexes have enhanced antibacterial and antifungal activities as compared to the free ligands.
Labeled vs. Label-Free Raman Imaging of Lipids in Endothelial Cells of Various Origins
Authors: Radwan Basseem , Adamczyk Adriana , Tott Szymon , Czamara Krzysztof , Kaminska Katarzyna , Matuszyk Ewelina , Baranska Malgorzata
Published: December 2020

Endothelial cells (EC) constitute a single layer of the lining of blood vessels and play an important role in maintaining cardiovascular homeostasis. Endothelial dysfunction has been recognized as a primary or secondary cause of many diseases and it manifests itself, among others, by increased lipid content or a change in the lipid composition in the EC. Therefore, the analysis of cellular lipids is crucial to understand the mechanisms of disease development. Tumor necrosis factor alpha (TNF-α)-induced inflammation of EC alters the lipid content of cells, which can be detected by Raman spectroscopy. By default, lipid detection is carried out in a label-free manner, and these compounds are recognized based on their spectral profile characteristics. We consider (3S,3′S)-astaxanthin (AXT), a natural dye with a characteristic resonance spectrum, as a new Raman probe for the detection of lipids in the EC of various vascular beds, i.e., the aorta, brain and heart. AXT colocalizes with lipids in cells, enabling imaging of lipid-rich cellular components in a time-dependent manner using laser power 10 times lower than that commonly used to measure biological samples. The results show that AXT can be used to study lipids distribution in EC at various locations, suggesting its use as a universal probe for studying cellular lipids using Raman spectroscopy. The use of labeled Raman imaging of lipids in the EC of various organs could contribute to their easier identification and to a better understanding of the development and progression of various vascular diseases, and it could also potentially improve their diagnosis and treatment.
The Callus of Phaseolus coccineus and Glycine max Biotransform Flavanones into the Corresponding Flavones
Authors: Dymarska Monika , Janeczko Tomasz , Kostrzewa-Susłow Edyta
Published: December 2020

In vitro plant cultures are gaining in industrial importance, especially as biocatalysts and as sources of secondary metabolites used in pharmacy. The idea that guided us in our research was to evaluate the biocatalytic potential of newly obtained callus tissue towards flavonoid compounds. In this publication, we describe new ways of using callus cultures in the biotransformations. In the first method, the callus cultures grown on a solid medium are transferred to the water, the reaction medium into which the substrate is introduced. In the second method, biotransformation is carried out on a solid medium by growing callus cultures. In the course of the research, we have shown that the callus obtained from Phaseolus coccineus and Glycine max is capable of converting flavanone, 5-methoxyflavanone and 6-methoxyflavanone into the corresponding flavones.
Ferula L. Plant Extracts and Dose-Dependent Activity of Natural Sesquiterpene Ferutinin: From Antioxidant Potential to Cytotoxic Effects
Authors: Macrì Roberta , Musolino Vincenzo , Gliozzi Micaela , Carresi Cristina , Maiuolo Jessica , Nucera Saverio , Scicchitano Miriam , Bosco Francesca , Scarano Federica , Ruga Stefano , Zito Maria Caterina , Guarnieri Lorenza , Bombardelli Ezio , Mollace Vincenzo
Published: December 2020

The employment studies of natural extracts in the prevention and treatment of several diseases highlighted the role of different species of genus Ferula L., belonging to the Apiaceae family, dicotyledonous plants present in many temperate zones of our planet. Ferula communis L. is the main source of sesquiterpene ferutinin, a bioactive compound studied both in vitro and in vivo, because of different effects, such as phytoestrogenic, antioxidant, anti-inflammatory, but also antiproliferative and cytotoxic activity, performed in a dose-dependent and cell-dependent way. The present review will focus on the molecular mechanisms involved in the different activities of Ferutinin, starting from its antioxidant potential at low doses until its ionophoric property and the subsequent mitochondrial dysfunction induced through administration of high doses, which represent the key point of its anticancer action. Furthermore, we will summarize the data acquired from some experimental studies on different cell types and on several diseases. The results obtained showed an important antioxidant and phytoestrogenic regulation with lack of typical side effects related to estrogenic therapy. The preferential cell death induction for tumor cell lines suggests that ferutinin may have anti-neoplastic properties, and may be used as an antiproliferative and cytotoxic agent in an estrogen dependent and independent manner. Nevertheless, more data are needed to clearly understand the effect of ferutinin in animals before using it as a phytoestrogen or anticancer drug.
Small Interfering RNAs Are Highly Effective Inhibitors of Crimean-Congo Hemorrhagic Fever Virus Replication In Vitro
Authors: Földes Fanni , Madai Mónika , Papp Henrietta , Kemenesi Gábor , Zana Brigitta , Geiger Lili , Gombos Katalin , Somogyi Balázs , Bock-Marquette Ildikó , Jakab Ferenc
Published: December 2020

Crimean-Congo hemorrhagic fever virus (CCHFV) is one of the prioritized diseases of the World Health Organization, considering its potential to create a public health emergency and, more importantly, the absence of efficacious drugs and/or vaccines for treatment. The highly pathogenic characteristic of CCHFV restricts research to BSL-4 laboratories, which complicates effective research and developmental strategies. In consideration of antiviral therapies, RNA interference can be used to suppress viral replication by targeting viral genes. RNA interference uses small interfering RNAs (siRNAs) to silence genes. The aim of our study was to design and test siRNAs in vitro that inhibit CCHFV replication and can serve as a basis for further antiviral therapies. A549 cells were infected with CCHFV after transfection with the siRNAs. Following 72 h, nucleic acid from the supernatant was extracted for RT Droplet Digital PCR analysis. Among the investigated siRNAs we identified effective candidates against all three segments of the CCHF genome. Consequently, blocking any segment of CCHFV leads to changes in the virus copy number that indicates an antiviral effect of the siRNAs. In summary, we demonstrated the ability of specific siRNAs to inhibit CCHFV replication in vitro. This promising result can be integrated into future anti-CCHFV therapy developments.
Simultaneous Ultrasound and Heat Enhance Functional Properties of Glycosylated Lactoferrin
Authors: Li Zhipeng , Ma Dexue , He Yiyang , Guo Siqi , Liu Fuguo , Liu Xuebo
Published: December 2020

Protein-polysaccharide covalent complexes exhibit better physicochemical and functional properties than single protein or polysaccharide. To promote the formation of the covalent complex from lactoferrin (LF) and beet pectin (BP), we enhanced the Maillard reaction between LF and BP by using an ultrasound-assisted treatment and studied the structure and functional properties of the resulting product. The reaction conditions were optimized by an orthogonal experimental design, and the highest grafting degree of 55.36% was obtained by ultrasonic treatment at 300 W for 20 min and at LF concentration of 20 g/L and BP concentration of 9 g/L. The formation of LF-BP conjugates was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) spectroscopy. Ultrasound-assisted treatment can increase the surface hydrophobicity, browning index, 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 2,2’-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radicals scavenging activity of LF due to the changes in the spatial configuration and formation of Maillard reaction products. The thermal stability, antioxidant activity and emulsifying property of LF were significantly improved after combining with BP. These findings reveal the potential application of modified proteins by ultrasonic and heat treatment.
Synthesis, Characterization and Biological Evaluation of Magnolol and Honokiol Derivatives with 1,3,5-Triazine of Metformin Cyclization
Authors: Ren Cui , Wang Juanxia , Tan Youzhen , Guo Mingxin , Guo Jieqing , Liu Ying , Wu Xia , Feng Yifan
Published: December 2020

Herein, we sought to evaluate the contribution of the 1,3,5-triazine ring through the metformin cyclization unit to the biological activity of magnolol and honokiol-conjugates. One of the phenolic OH groups of magnolol or honokiol was replaced by a 1,3,5-triazine ring to further explore their synthesis and medicinal versatility. In this study, a robust procedure of three steps was adopted for the synthesis of magnolol and honokiol derivatives by alkylation of potassium carbonate with a 1,3,5-triazine ring. To our knowledge, this is the first report to connect one of the phenolic OH positions of magnolol or honokiol to a 1,3,5-triazine ring cyclized by metformin. The structural characterization of three new compounds was carried out via spectroscopic techniques, i.e., 13C NMR, 1H NMR, and HRMS. Surprisingly, these compounds showed no cytotoxicity against RAW 264.7 macrophages but significantly inhibited the proliferation of MCF-7 (human breast cancer cells), HepG2 (human hepatoma cells), A549 (human lung carcinoma cells), and BxPC-3 (human pancreatic carcinoma cells) tumor cell lines. Furthermore, the compounds also significantly inhibited the release of inflammatory cytokines, including nitric oxide (NO), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in the lipopolysaccharide (LPS)-activated mouse cells (RAW 264.7). Among them, compound 2 demonstrated promising broad-spectrum antiproliferative potential with half inhibitory concentration (IC50) values ranging from 5.57 to 8.74 µM and it significantly decreased caspase-3 and Bcl-2 expression in HepG2 cells. These interesting findings show that derivatization of magnolol and honokiol with 1,3,5-triazine affects and modulates their biological properties.
New Design Method for Fabricating Multilayer Membranes Using CO2-Assisted Polymer Compression Process
Authors: Aizawa Takafumi
Published: December 2020

It was verified that deep learning can be used in creating multilayer membranes with multiple porosities using the CO2-assisted polymer compression (CAPC) method. To perform training while reducing the number of experimental data as much as possible, the experimental data of the compression behavior of two layers were expanded to three layers for training, but sufficient accuracy could not be obtained. However, the accuracy was dramatically improved by adding the experimental data of the three layers. The possibility of only simulating process results without the necessity for a model is a merit unique to deep learning. Overall, in this study, the results show that by devising learning data, deep learning is extremely effective in designing multilayer membranes using the CAPC method.
Modern Trends in the In Vitro Production and Use of Callus, Suspension Cells and Root Cultures of Medicinal Plants
Authors: Babich Olga , Sukhikh Stanislav , Pungin Artem , Ivanova Svetlana , Asyakina Lyudmila , Prosekov Alexander
Published: December 2020

This paper studies modern methods of producing and using callus, suspension cells and root cultures of medicinal plants in vitro. A new solution for natural product production is the use of an alternative source of renewable, environmentally friendly raw materials: callus, suspension and root cultures of higher plants in vitro. The possibility of using hairy root cultures as producers of various biologically active substances is studied. It is proven that the application of the genetic engineering achievements that combine in vitro tissue culture and molecular biology methods was groundbreaking in terms of the intensification of the extraction process of compounds significant for the medical industry. It is established that of all the callus processing methods, suspension and root cultures in vitro, the Agrobacterium method is the most widely used in practice. The use of agrobacteria has advantages over the biolistic method since it increases the proportion of stable transformation events, can deliver large DNA segments and does not require special ballistic devices. As a result of the research, the most effective strains of agrobacteria are identified.
The Effects of Prolonged Storage on ARPE-19 Cells Stored at Three Different Storage Temperatures
Authors: Islam Rakibul , Corraya Rima Maria , Pasovic Lara , Khan Ayyad Zartasht , Aass Hans Christian D. , Eidet Jon Roger , Utheim Tor Paaske
Published: December 2020

This study aimed to investigate how prolonged storage of adult retinal pigment epithelial (ARPE-19) cell sheets affects cell metabolism, morphology, viability, and phenotype. ARPE-19 cell sheets were stored at three temperatures (4 °C, 16 °C, and 37 °C) for three weeks. Metabolic status and morphology of the cells were monitored by sampling medium and examining cells by phase-contrast microscopy, respectively, throughout the storage period. Cell viability was analyzed by flow cytometry, and phenotype was determined by epifluorescence microscopy after the storage. Lactate production and glucose consumption increased heavily, while pH dropped considerably, through storage at 37 °C compared to 4 °C and 16 °C. During storage, morphology started to deteriorate first at 4 °C, then at 37 °C, and was maintained the longest at 16 °C. Viability of the cells after three weeks of storage was best preserved at 16 °C, while cells stored at 4 °C and 37 °C had reduced viability. Dedifferentiation indicated by reduced expression of retinal pigment epithelium-specific protein 65 (RPE65), zonula occludens protein 1 (ZO-1), and occludin after three weeks of storage was noticed in all experimental groups compared to control. We conclude that storage temperature affects the metabolic status of ARPE-19 cells and that 16 °C reduces metabolic activity while protecting viability and morphology.
New Insights for Red Propolis of Alagoas—Chemical Constituents, Topical Membrane Formulations and Their Physicochemical and Biological Properties
Authors: Silva Valdemir C. , Silva Abiane M. G. S. , Basílio Jacqueline A. D. , Xavier Jadriane A. , do Nascimento Ticiano G. , Naal Rose M. Z. G. , del Lama Maria Perpetua , Leonelo Laila A. D. , Mergulhão Naianny L. O. N. , Maranhão Fernanda C. A. , Silva Denise M. W. , Owen Robert , Duarte Ilza F. B. , Bulhões Laisa C. G. , Basílio Irinaldo D. , Goulart Marília O. F.
Published: December 2020

The main objectives of this study were to evaluate the chemical constitution and allergenic potential of red propolis extract (RPE). They were evaluated, using high performance liquid chromatography (HPLC) and the release of β-hexosaminidase, respectively. A plethora of biologically active polyphenols and the absence of allergic responses were evinced. RPE inhibited the release of β-hexosaminidase, suggesting that the extract does not stimulate allergic responses. Additionally, the physicochemical properties and antibacterial activity of hydrogel membranes loaded with RPE were analyzed. Bio-polymeric hydrogel membranes (M) were obtained using 5% carboxymethylcellulose (M1 and M2), 1.0% of citric acid (M3) and 10% RPE (for all). Their characterization was performed using thermal analysis, Fourier transform infrared (FTIR), total phenolic content, phenol release test and, antioxidant activity through 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and Ferric Reducing Antioxidant Power (FRAP). The latter appointed to the similar antioxidant capacity of the M1, M2 and M3. The degradation profiles showed higher thermostability to M3, followed by M2 and M1. The incorporation of RPE into the matrices and the crosslinking of M3 were evinced by FTIR. There were differences in the release of phenolic compounds, with a higher release related to M1 and lower in the strongly crosslinked M3. The degradation profiles showed higher thermostability to M3, followed by M2 and M1. The antibacterial activity of the membranes was determined using the disc diffusion assay, in comparison with controls, obtained in the same way, without RPE. The membranes elicited antibacterial activity against Staphylococcus aureus and Staphylococcus epidermidis, with superior performance over M3. The hydrogel membranes loaded with RPE promote a physical barrier against bacterial skin infections and may be applied in the wound healing process.
Zeaxanthin, a Molecule for Photoprotection in Many Different Environments
Authors: Demmig-Adams Barbara , Stewart Jared J. , López-Pozo Marina , Polutchko Stephanie K. , Adams William W.
Published: December 2020

Conversion of sunlight into photochemistry depends on photoprotective processes that allow safe use of sunlight over a broad range of environmental conditions. This review focuses on the ubiquity of photoprotection associated with a group of interconvertible leaf carotenoids, the xanthophyll cycle. We survey the striking plasticity of this process observed in nature with respect to (1) xanthophyll cycle pool size, (2) degree and speed of interconversion of its components, and (3) flexibility in the association between xanthophyll cycle conversion state and photoprotective dissipation of excess excitation energy. It is concluded that the components of this system can be independently tuned with a high degree of flexibility to produce a fit for different environments with various combinations of light, temperature, and other factors. In addition, the role of genetic variation is apparent from variation in the response of different species growing side-by-side in the same environment. These findings illustrate how field studies can generate insight into the adjustable levers that allow xanthophyll cycle-associated photoprotection to support plant photosynthetic productivity and survival in environments with unique combinations of environmental factors.
How the Supramolecular Nature of Lignohumate Affects Its Diffusion in Agarose Hydrogel
Authors: Klučáková Martina , Kalina Michal , Enev Vojtěch
Published: December 2020

Lignohumate, as an industrially produced analog of natural humic substances, is studied from the point of view of its diffusion properties. This work focuses on its permeation ability, important in agricultural and horticultural applications, connected with its penetration into plant organs as leaves and roots. The hydrogel based on agarose was used as a model material for the diffusion of lignohumate. Two types of experiments were realized: the diffusion of lignohumate in the hydrogel diffusion couple and the diffusion of lignohumate from its solution into hydrogel. The diffusion coefficient of lignohumate in the hydrogel was determined and used for the modelling of the time development of concentration profiles. It was found that the model agrees with experimental data for short times but an accumulation of lignohumate in front of the interface between donor and acceptor hydrogels was observed after several days. The particle size distribution of lignohumate and changes in the E4/E6 ratio used as an indicator of molecular weight of humic substances were determined. The results showed that the supramolecular structure of lignohumate can react sensitively to actual changes in its environs and thus affect their mobility and permeability into different materials. A filtration effect at the interface can be observed as an accompanying phenomenon of the re-arrangement in the lignohumate secondary structure.
Nitro-, Cyano-, and Methylfuroxans, and Their Bis-Derivatives: From Green Primary to Melt-Cast Explosives
Authors: Larin Alexander A. , Bystrov Dmitry M. , Fershtat Leonid L. , Konnov Alexey A. , Makhova Nina N. , Monogarov Konstantin A. , Meerov Dmitry B. , Melnikov Igor N. , Pivkina Alla N. , Kiselev Vitaly G. , Muravyev Nikita V.
Published: December 2020

In the present work, we studied in detail the thermochemistry, thermal stability, mechanical sensitivity, and detonation performance for 20 nitro-, cyano-, and methyl derivatives of 1,2,5-oxadiazole-2-oxide (furoxan), along with their bis-derivatives. For all species studied, we also determined the reliable values of the gas-phase formation enthalpies using highly accurate multilevel procedures W2-F12 and/or W1-F12 in conjunction with the atomization energy approach and isodesmic reactions with the domain-based local pair natural orbital (DLPNO) modifications of the coupled-cluster techniques. Apart from this, we proposed reliable benchmark values of the formation enthalpies of furoxan and a number of its (azo)bis-derivatives. Additionally, we reported the previously unknown crystal structure of 3-cyano-4-nitrofuroxan. Among the monocyclic compounds, 3-nitro-4-cyclopropyl and dicyano derivatives of furoxan outperformed trinitrotoluene, a benchmark melt-cast explosive, exhibited decent thermal stability (decomposition temperature >200 °C) and insensitivity to mechanical stimuli while having notable volatility and low melting points. In turn, 4,4′-azobis-dicarbamoyl furoxan is proposed as a substitute of pentaerythritol tetranitrate, a benchmark brisant high explosive. Finally, the application prospects of 3,3′-azobis-dinitro furoxan, one of the most powerful energetic materials synthesized up to date, are limited due to the tremendously high mechanical sensitivity of this compound. Overall, the investigated derivatives of furoxan comprise multipurpose green energetic materials, including primary, secondary, melt-cast, low-sensitive explosives, and an energetic liquid.
Implication of Lactucopicrin in Autophagy, Cell Cycle Arrest and Oxidative Stress to Inhibit U87Mg Glioblastoma Cell Growth
Authors: Rotondo Rossella , Oliva Maria Antonietta , Staffieri Sabrina , Castaldo Salvatore , Giangaspero Felice , Arcella Antonietta
Published: December 2020

In this study, we propose lactucopicrin (LCTP), a natural sesquiterpene lactone from Lactucavirosa, as a molecule able to control the growth of glioblastoma continuous cell line U87Mg. The IC50 of U87Mg against LCTP revealed a strong cytotoxic effect. Daily administration of LCTP showed a dose and time-dependent reduction of GBM cell growth and viability, also confirmed by inhibition of clonogenic potential and mobility of U87Mg cells. LCTP activated autophagy in U87Mg cells and decreased the phosphorylation of proliferative signals pAKT and pERK. LCTP also induced the cell cycle arrest in G2/M phase, confirmed by decrease of CDK2 protein and increase of p53 and p21. LCTP stimulated apoptosis as evidenced by reduction of procaspase 6 and the increase of the cleaved/full-length PARP ratio. The pre-treatment of U87Mg cells with ROS scavenger N-acetylcysteine (NAC), which reversed its cytotoxic effect, showed the involvement of LCTP in oxidative stress. Finally, LCTP strongly enhanced the sensitivity of U87Mg cells to canonical therapy Temozolomide (TMZ) and synergized with this drug. Altogether, the growth inhibition of U87Mg GBM cells induced by LCTP is the result of several synergic mechanisms, which makes LCTP a promising adjuvant therapy for this complex pathology.
Novel 3-Acetyl-2,5-disubstituted-1,3,4-oxadiazolines: Synthesis and Biological Activity
Authors: Paruch Kinga , Popiołek Łukasz , Biernasiuk Anna , Hordyjewska Anna , Malm Anna , Wujec Monika
Published: December 2020

The aim of our study was the two-stage synthesis of 1,3,4-oxadiazole derivatives. The first step was the synthesis of hydrazide–hydrazones from 3-methyl-4-nitrobenzhydrazide and the corresponding substituted aromatic aldehydes. Then, the synthesized hydrazide–hydrazones were cyclized with acetic anhydride to obtain new 3-acetyl-2,3-disubstituted-1,3,4-oxadiazolines. All of obtained compounds were tested in in vitro assays to establish their potential antimicrobial activity and cytotoxicity. Our results indicated that few of the newly synthesized compounds had some antimicrobial activity, mainly compounds 20 and 37 towards all used reference bacterial strains (except Klebsiella pneumoniae, Proteus mirabilis, and Pseudomonas aeruginosa) and fungi. These substances showed a strong or powerful bactericidal effect, especially against Staphylococcus spp. belonging to Gram-positive bacteria. Compound 37 was active against Staphylococcus epidermidis at minimal inhibitory concentration (MIC) = 0.48 µg/mL and was characterized by low cytotoxicity. This compound possessed quinolin-4-yl substituent in the second position of 1,3,4-oxadiazole ring and 3-methyl-4-nitrophenyl in position 5. High effectiveness and safety of these derivatives make them promising candidates as antimicrobial agents. Whereas the compound 20 with the 5-iodofurane substituent in position 2 of the 1,3,4-oxadiazole ring showed the greatest activity against S. epidermidis at MIC = 1.95 µg/mL.
Blockade of Catecholamine Reuptake in the Prelimbic Cortex Decreases Top-Down Attentional Control in Response to Novel, but not Familiar Appetitive Distracters, within a Timing Paradigm
Authors: Matthews Alexander R. , Buhusi Mona , Buhusi Catalin V.
Published: December 2020

Emotionally charged distracters delay timing behavior. Increasing catecholamine levels within the prelimbic cortex has beneficial effects on timing by decreasing the delay after aversive distracters. Here we examined whether increasing catecholamine levels within the prelimbic cortex also protects against the deleterious timing delays caused by novel distracters or by familiar appetitive distracters. Rats were trained in a peak-interval procedure and tested in trials with either a novel (unreinforced) distracter, a familiar appetitive (food-reinforced) distracter, or no distracter after being locally infused with catecholamine reuptake blocker nomifensine within the prelimbic cortex. Prelimbic infusion of nomifensine did not alter timing accuracy and precision. However, it increased the delay caused by novel distracters in an inverted-U dose-dependent manner, while being ineffective for appetitive distracters. Together with previous data, these results suggest that catecholaminergic modulation of prelimbic top-down attentional control of interval timing varies with distracter’s valence: prelimbic catecholamines increase attentional control when presented with familiar aversive distracters, have no effect on familiar neutral or familiar appetitive distracters, and decrease it when presented with novel distracters. These findings detail complex interactions between catecholaminergic modulation of attention to timing and nontemporal properties of stimuli, which should be considered when developing therapeutic methods for attentional or affective disorders.
A Literature Review of Attentional Biases amongst Individuals with Substance Dependency: Individual Differences and Modulating Factors
Authors: Zhang Melvyn , Fung Daniel S.S. , Smith Helen
Published: December 2020

Introduction: Advances in experimental psychology have led to a better understanding of automatic, unconscious processes, referred to as attentional biases. Despite the growing evidence from meta-analytical studies, we still do not understand why some individuals have a greater magnitude of these biases, and why others have none. There has been little focus on elucidating individual differences and task parameters that affect the overall magnitude of the biases. In this opinion piece, we will attempt to identify these. We will then discuss both the research and clinical implications. Methods and Analysis: To identify the factors that modulated the magnitude of attentional biases across all the substance disorders (i.e., opioid use, cannabis use, and stimulant-use disorders), we performed a search using the bibliographic databases PubMed and MEDLINE. The search terminologies “attention bias” or “cognitive bias” or “approach bias” or “avoidance bias” were used when we looked for relevant articles. Results: It was evident from the published literature that several individual differences and factors modulated the magnitude of baseline biases. Across opioid, cannabis, and stimulant-use disorders, the most common individual differences identified were the severity of the dependence and the quantity of substance used. For both opioid and cannabis disorders the timing of stimulus presentation influenced the detection of attentional bias; it appeared that short stimulus timing was better able to detect attentional bias. Other identified individual differences included subjective craving and impulsiveness. The results highlight several research and clinical implications. Conclusions: The discovery of these individual differences and factors of the task paradigm that affect the magnitude of attentional biases will help in the future conceptualization of attention-bias-modification intervention.
Association of Mediterranean Diet Adherence, Functional Food Consumption and Anthropometric Characteristics with Anxiety and Depression Indexes in a Sample of Healthy Greek Adults: A Cross-Sectional Study
Authors: Elmaliklis Ioannis-Nektarios , Miserli Elisavet , Filipatou Maria , Tsikouras Ioannis , Dimou Charalampia , Koutelidakis Antonios
Published: December 2020

The purpose of this randomized, retrospective study was to investigate the possible association among levels of anxiety and depression and functional food consumption frequency, Mediterranean diet adoption and anthropometric indices in a sample of Greek, mainly young, adults. One hundred twenty healthy adults, 80% of whom were 18–35 years old, were randomly recruited from the North Aegean Islands, mainly from the Aegean University campus. The degree of anxiety was evaluated using the State-Trait Anxiety Inventory (STAI-X-1,2) whereas the degree of depression was assessed using the Beck Depression Inventory (BDI). Mediterranean diet adherence was evaluated using the Med Diet Score and functional foods consumption frequency by a validated food frequency questionnaire (FFQ). According to our results, the study did not show a statistically significant association between Mediterranean diet adherence and anxiety or depression levels (p > 0.05), and a trend association was observed only for the depression grouped score. In parallel, increased depression level was significantly correlated with an increased body mass index (BMI) (p = 0.019). In addition, the consumption of specific functional foods, such as beans, nuts, pomegranate, spinach and foods enriched with β-glucans, correlated with state-trait anxiety or depression levels, as well as with anthropometric indices (p < 0.05). The results of the study may contribute to the elucidation of the possible role of the Mediterranean diet and functional food consumption on self-esteem, anxiety and depression.
Implementation of a Lightweight Semantic Segmentation Algorithm in Road Obstacle Detection
Authors: Liu Bushi , Lv Yongbo , Gu Yang , Lv Wanjun
Published: December 2020

Due to deep learning’s accurate cognition of the street environment, the convolutional neural network has achieved dramatic development in the application of street scenes. Considering the needs of autonomous driving and assisted driving, in a general way, computer vision technology is used to find obstacles to avoid collisions, which has made semantic segmentation a research priority in recent years. However, semantic segmentation has been constantly facing new challenges for quite a long time. Complex network depth information, large datasets, real-time requirements, etc., are typical problems that need to be solved urgently in the realization of autonomous driving technology. In order to address these problems, we propose an improved lightweight real-time semantic segmentation network, which is based on an efficient image cascading network (ICNet) architecture, using multi-scale branches and a cascaded feature fusion unit to extract rich multi-level features. In this paper, a spatial information network is designed to transmit more prior knowledge of spatial location and edge information. During the course of the training phase, we append an external loss function to enhance the learning process of the deep learning network system as well. This lightweight network can quickly perceive obstacles and detect roads in the drivable area from images to satisfy autonomous driving characteristics. The proposed model shows substantial performance on the Cityscapes dataset. With the premise of ensuring real-time performance, several sets of experimental comparisons illustrate that SP-ICNet enhances the accuracy of road obstacle detection and provides nearly ideal prediction outputs. Compared to the current popular semantic segmentation network, this study also demonstrates the effectiveness of our lightweight network for road obstacle detection in autonomous driving.