bluuebunny/crossref_metadata_2025 huggingface dataset:
Fraxetin Suppresses Cell Proliferation and Induces Apoptosis through Mitochondria Dysfunction in Human Hepatocellular Carcinoma Cell Lines Huh7 and Hep3B
Authors: Song Jisoo , Ham Jiyeon , Hong Taeyeon , Song Gwonhwa , Lim Whasun
Published: January 2021

Fraxetin is a coumarin scaffold compound extracted from Fraxinus rhynchophylla. It has antioxidant, anti-inflammatory, hepatoprotective, and antifibrotic effects. Furthermore, fraxetin has anticancer effects in breast and lung cancer. We aimed to evaluate whether fraxetin has anticancer activity in hepatocellular carcinoma (HCC) cells and its underlying mechanism. We demonstrated the anticancer effects of fraxetin in the HCC cell lines Huh7 and Hep3B. We confirmed that fraxetin inhibited cell proliferation (42% ± 10% Huh7; 52% ± 7% Hep3B) by arresting the cell cycle and inducing apoptosis in both cell lines. Moreover, fraxetin increased reactive oxygen species production (221% ± 55% Huh7; 460% ± 73% Hep3B), depolarized the mitochondrial membranes (ΔΨm) (345% ± 160% Huh7; 462% ± 140% Hep3B), and disrupted calcium homeostasis in both HCC cell lines. Chelating calcium ions with BAPTA-AM restored proliferation in fraxetin-treated Huh7 cells but not in Hep3B cells. Fraxetin did not affect the phosphorylation of extracellular-signal-regulated kinase 1/2, whereas it decreased JNK and phosphoinositide 3-kinase signaling. Furthermore, fraxetin and mitogen-activated protein kinase pharmacological inhibitors had synergistic antiproliferative effects on HCC cells. Although our study was limited to in vitro data that require validation, we suggest that fraxetin is a potential therapeutic agent against HCC progression.
Multiple-Monitor HPLC Assays for Rapid Process Development, In-Process Monitoring, and Validation of AAV Production and Purification
Authors: Gagnon Pete , Goricar Blaz , Mencin Nina , Zvanut Timotej , Peljhan Sebastijan , Leskovec Maja , Strancar Ales
Published: January 2021

HPLC is established as a fast convenient analytical technology for characterizing the content of empty and full capsids in purified samples containing adeno-associated virus (AAV). UV-based monitoring unfortunately over-estimates the proportion of full capsids and offers little value for characterizing unpurified samples. The present study combines dual-wavelength UV monitoring with intrinsic fluorescence, extrinsic fluorescence, and light-scattering to extend the utility of HPLC for supporting development of therapeutic AAV-based drugs. Applications with anion exchange (AEC), cation exchange (CEC), and size exclusion chromatography (SEC) are presented. Intrinsic fluorescence increases sensitivity of AAV detection over UV and enables more objective estimation of empty and full capsid ratios by comparison of their respective peak areas. Light scattering enables identification of AAV capsids in complex samples, plus semiquantitative estimation of empty and full capsid ratios from relative peak areas of empty and full capsids. Extrinsic Picogreen fluorescence enables semiquantitative tracking of DNA with all HPLC methods at all stages of purification. It does not detect encapsidated DNA but reveals DNA associated principally with the exteriors of empty capsids. It also enables monitoring of host DNA contamination across chromatograms. These enhancements support many opportunities to improve characterization of raw materials and process intermediates, to accelerate process development, provide rapid in-process monitoring, and support process validation.
Complex Factors and Challenges that Affect the Pharmacology, Safety and Efficacy of Nanocarrier Drug Delivery Systems
Authors: Piscatelli Joseph A. , Ban Jisun , Lucas Andrew T. , Zamboni William C.
Published: January 2021

Major developments in nanomedicines, such as nanoparticles (NPs), nanosomes, and conjugates, have revolutionized drug delivery capabilities over the past four decades. Although nanocarrier agents provide numerous advantages (e.g., greater solubility and duration of systemic exposure) compared to their small-molecule counterparts, there is considerable inter-patient variability seen in the systemic disposition, tumor delivery and overall pharmacological effects (i.e., anti-tumor efficacy and unwanted toxicity) of NP agents. This review aims to provide a summary of fundamental factors that affect the disposition of NPs in the treatment of cancer and why they should be evaluated during preclinical and clinical development. Furthermore, this chapter will highlight some of the translational challenges associated with elements of NPs and how these issues can only be addressed by detailed and novel pharmacology studies.
Suppression of Peritoneal Fibrosis by Sonoporation of Hepatocyte Growth Factor Gene-Encoding Plasmid DNA in Mice
Authors: Nishimura Koyo , Ogawa Koki , Kawaguchi Maho , Fumoto Shintaro , Mukai Hidefumi , Kawakami Shigeru
Published: January 2021

Gene therapy is expected to be used for the treatment of peritoneal fibrosis, which is a serious problem associated with long-term peritoneal dialysis. Hepatocyte growth factor (HGF) is a well-known anti-fibrotic gene. We developed an ultrasound and nanobubble-mediated (sonoporation) gene transfection system, which selectively targets peritoneal tissues. Thus, we attempted to treat peritoneal fibrosis by sonoporation-based human HGF (hHGF) gene transfection in mice. To prepare a model of peritoneal fibrosis, mice were intraperitoneally injected with chlorhexidine digluconate. We evaluated the preventive and curative effects of sonoporation-based hHGF transfection by analyzing the following factors: hydroxyproline level, peritoneum thickness, and the peritoneal equilibration test. The transgene expression characteristics of sonoporation were also evaluated using multicolor deep imaging. In early-stage fibrosis in mice, transgene expression by sonoporation was observed in the submesothelial layer. Sonoporation-based hHGF transfection showed not only a preventive effect but also a curative effect for early-stage peritoneal fibrosis. Sonoporation-based hHGF transfection may be suitable for the treatment of peritoneal fibrosis regarding the transfection characteristics of transgene expression in the peritoneum under fibrosis.
Lipophilic Peptide Dendrimers for Delivery of Splice-Switching Oligonucleotides
Authors: Daralnakhla Haneen , Saher Osama , Zamolo Susanna , Bazaz Safa , P. Bost Jeremy , Heitz Marc , Lundin Karin E. , EL Andaloussi Samir , Darbre Tamis , Reymond Jean-Louis , Zain Rula , Smith C. I. Edvard
Published: January 2021

Non-viral transfection reagents are continuously being developed in attempt to replace viral vectors. Among those non-viral vectors, dendrimers have gained increasing interest due to their unique molecular structure and multivalency. However, more improvements are still needed to achieve higher efficacy and lower toxicity. In this study, we have examined 18 peptide dendrimers conjugated to lipophilic moieties, such as fatty acids or hydrophobic amino acids, that were previously explored for siRNA. Reporter cells were employed to investigate the transfection of single strand splice-switching oligonucleotides (ONs) using these peptide dendrimers. Luciferase level changes reflecting efficiency varied with amino acid composition, stereochemistry, and complexation media used. 3rd generation peptide dendrimers with D-amino acid configuration were superior to L-form. Lead formulations with 3rd generation, D-amino acid peptide dendrimers increased the correction level of the delivered ON up to 93-fold over untreated HeLa Luc/705 cells with minimal toxicity. To stabilize the formed complexes, Polyvinyl alcohol 18 (PVA18) polymer was added. Although PVA18 addition increased activity, toxicity when using our best candidates G 2,3KL-(Leu)4 (D) and G 2,3KL-diPalmitamide (D) was observed. Our findings demonstrate the potential of lipid-conjugated, D-amino acid-containing peptide dendrimers to be utilized as an effective and safe delivery vector for splice-switching ONs.
Erratum: Sanz del Olmo, N.; et al. Antioxidant and Antibacterial Properties of Carbosilane Dendrimers Functionalized with Polyphenolic Moieties. Pharmaceutics 2020, 12, 698
Authors: Sanz del Olmo Natalia , Peña González Cornelia E. , Rojas Daniel , Gómez Rafael , Ortega Paula , Escarpa Alberto , de la Mata Francisco Javier
Published: January 2021

Due to an error during production, in [...]
Unlocking the Power of Exosomes for Crossing Biological Barriers in Drug Delivery
Authors: Elliott Rebekah Omarkhail , He Mei
Published: January 2021

Since the 2013 Nobel Prize was awarded for the discovery of vesicle trafficking, a subgroup of nanovesicles called exosomes has been driving the research field to a new regime for understanding cellular communication. This exosome-dominated traffic control system has increased understanding of many diseases, including cancer metastasis, diabetes, and HIV. In addition to the important diagnostic role, exosomes are particularly attractive for drug delivery, due to their distinctive properties in cellular information transfer and uptake. Compared to viral and non-viral synthetic systems, the natural, cell-derived exosomes exhibit intrinsic payload and bioavailability. Most importantly, exosomes easily cross biological barriers, obstacles that continue to challenge other drug delivery nanoparticle systems. Recent emerging studies have shown numerous critical roles of exosomes in many biological barriers, including the blood–brain barrier (BBB), blood–cerebrospinal fluid barrier (BCSFB), blood–lymph barrier (BlyB), blood–air barrier (BAB), stromal barrier (SB), blood–labyrinth barrier (BLaB), blood–retinal barrier (BRB), and placental barrier (PB), which opens exciting new possibilities for using exosomes as the delivery platform. However, the systematic reviews summarizing such discoveries are still limited. This review covers state-of-the-art exosome research on crossing several important biological barriers with a focus on the current, accepted models used to explain the mechanisms of barrier crossing, including tight junctions. The potential to design and engineer exosomes to enhance delivery efficacy, leading to future applications in precision medicine and immunotherapy, is discussed.
Potential Herb–Drug Interactions in the Management of Age-Related Cognitive Dysfunction
Authors: Auxtero Maria D. , Chalante Susana , Abade Mário R. , Jorge Rui , Fernandes Ana I.
Published: January 2021

Late-life mild cognitive impairment and dementia represent a significant burden on healthcare systems and a unique challenge to medicine due to the currently limited treatment options. Plant phytochemicals have been considered in alternative, or complementary, prevention and treatment strategies. Herbals are consumed as such, or as food supplements, whose consumption has recently increased. However, these products are not exempt from adverse effects and pharmacological interactions, presenting a special risk in aged, polymedicated individuals. Understanding pharmacokinetic and pharmacodynamic interactions is warranted to avoid undesirable adverse drug reactions, which may result in unwanted side-effects or therapeutic failure. The present study reviews the potential interactions between selected bioactive compounds (170) used by seniors for cognitive enhancement and representative drugs of 10 pharmacotherapeutic classes commonly prescribed to the middle-aged adults, often multimorbid and polymedicated, to anticipate and prevent risks arising from their co-administration. A literature review was conducted to identify mutual targets affected (inhibition/induction/substrate), the frequency of which was taken as a measure of potential interaction. Although a limited number of drugs were studied, from this work, interaction with other drugs affecting the same targets may be anticipated and prevented, constituting a valuable tool for healthcare professionals in clinical practice.
Assessments of the In Vitro and In Vivo Linker Stability and Catabolic Fate for the Ortho Hydroxy-Protected Aryl Sulfate Linker by Immuno-Affinity Capture Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometric Assay
Authors: Lee Byeong ill , Park Seo-jin , Park Yuri , Shin Seok-Ho , Choi Jang-mi , Park Min-jae , Lim Jeong-hyeon , Kim Sun Young , Lee Hyangsook , Shin Young G.
Published: January 2021

Antibody–drug conjugate (ADC) linkers play an important role in determining the safety and efficacy of ADC. The Ortho Hydroxy-Protected Aryl Sulfate (OHPAS) linker is a newly developed linker in the form of a di-aryl sulfate structure consisting of phenolic payload and self-immolative group (SIG). In this study, using two bioanalytical approaches (namely “bottom-up” and “middle-up” approaches) via the liquid chromatography-quadrupole time-of-flight mass spectrometric (LC-qTOF-MS) method, in vitro and in vivo linker stability experiments were conducted for the OHPAS linker. For comparison, the valine-citrulline-p-aminobenzyloxycarbonyl (VC-PABC) linker was also evaluated under the same experimental conditions. In addition, the catabolite identification experiments at the subunit intact protein level were simultaneously performed to evaluate the catabolic fate of ADCs. As a result, the OHPAS linker was stable in the in vitro mouse/human plasma as well as in vivo pharmacokinetic studies in mice, whereas the VC-PABC linker was relatively unstable in mice in vitro and in vivo. This is because the VC-PABC linker was sensitive to a hydrolytic enzyme called carboxylesterase 1c (Ces1c) in mouse plasma. In conclusion, the OHPAS linker appears to be a good linker for ADC, and further experiments would be warranted to demonstrate the efficacy and toxicity related to the OHPAS linker.
Anionic Complex with Efficient Expression and Good Safety Profile for mRNA Delivery
Authors: Hamada Eri , Kurosaki Tomoaki , Hashizume Junya , Harasawa Hitomi , Nakagawa Hiroo , Nakamura Tadahiro , Kodama Yukinobu , Sasaki Hitoshi
Published: January 2021

We previously found that a complex comprising plasmid DNA (pDNA), polyethylenimine (PEI), and γ-polyglutamic acid (γ-PGA) had high transgene efficiency without cytotoxicity in vitro and in vivo. However, messenger RNA (mRNA) remains an attractive alternative to pDNA. In this study, we developed a safe and effective delivery system for mRNA to prevent its degradation and efficiently deliver it into target cells. Various cationic and anionic complexes were produced containing PEI, γ-PGA, and an mRNA encoding firefly luciferase. Their physicochemical properties and cytotoxicities were analyzed and the in vitro and in vivo protein expression were determined. The cationic mRNA/PEI complex showed high in vitro protein expression with strong cytotoxicity. The anionic complex was constructed as mRNA/PEI8/γ-PGA12 complex with a theoretical charge ratio of 1:8:12 based on the phosphate groups of the mRNA, nitrogen groups of PEI, and carboxylate groups of γ-PGA. It was stable and showed high in vitro protein expression without cytotoxicity. After intravenous administration of mRNA/PEI8/γ-PGA12 complex to mice, high protein expression was observed in the spleen and liver and slight expression was observed in the lung over 24 h. Thus, the newly constructed mRNA/PEI8/γ-PGA12 complex provides a safe and effective strategy for the delivery of mRNA.
Molecular Mechanisms of Melatonin-Mediated Cell Protection and Signaling in Health and Disease
Authors: Kopustinskiene Dalia M. , Bernatoniene Jurga
Published: January 2021

Melatonin, an endogenously synthesized indolamine, is a powerful antioxidant exerting beneficial action in many pathological conditions. Melatonin protects from oxidative stress in ischemic/reperfusion injury, neurodegenerative diseases, and aging, decreases inflammation, modulates the immune system, inhibits proliferation, counteracts the Warburg effect, and promotes apoptosis in various cancer models. Melatonin stimulates antioxidant enzymes in the cells, protects mitochondrial membrane phospholipids, especially cardiolipin, from oxidation thus preserving integrity of the membranes, affects mitochondrial membrane potential, stimulates activity of respiratory chain enzymes, and decreases the opening of mitochondrial permeability transition pore and cytochrome c release. This review will focus on the molecular mechanisms of melatonin effects in the cells during normal and pathological conditions and possible melatonin clinical applications.
Comparison of Genetically Engineered Immunodeficient Animal Models for Nonclinical Testing of Stem Cell Therapies
Authors: Kim Yoon-Young , Kim Jin-Soo , Che Jeong-Hwan , Ku Seung-Yup , Kang Byeong-Cheol , Yun Jun-Won
Published: January 2021

For the recovery or replacement of dysfunctional cells and tissue—the goal of stem cell research—successful engraftment of transplanted cells and tissues are essential events. The event is largely dependent on the immune rejection of the recipient; therefore, the immunogenic evaluation of candidate cells or tissues in immunodeficient animals is important. Understanding the immunodeficient system can provide insights into the generation and use of immunodeficient animal models, presenting a unique system to explore the capabilities of the innate immune system. In this review, we summarize various immunodeficient animal model systems with different target genes as valuable tools for biomedical research. There have been numerous immunodeficient models developed by different gene defects, resulting in many different features in phenotype. More important, mice, rats, and other large animals exhibit very different immunological and physiological features in tissue and organs, including genetic background and a representation of human disease conditions. Therefore, the findings from this review may guide researchers to select the most appropriate immunodeficient strain, target gene, and animal species based on the research type, mutant gene effects, and similarity to human immunological features for stem cell research.
Bacterial-Derived Outer Membrane Vesicles are Potent Adjuvants that Drive Humoral and Cellular Immune Responses
Authors: Prior J. Timothy , Davitt Christopher , Kurtz Jonathan , Gellings Patrick , McLachlan James B. , Morici Lisa A.
Published: January 2021

Discovery and development of novel adjuvants that can improve existing or next generation vaccine platforms have received considerable interest in recent years. In particular, adjuvants that can elicit both humoral and cellular immune responses would be particularly advantageous because the majority of licensed vaccines are formulated with aluminum hydroxide (alum) which predominantly promotes antibodies. We previously demonstrated that bacterial-derived outer membrane vesicles (OMV) possess inherent adjuvanticity and drive antigen-specific antibody and cellular immune responses to OMV components. Here, we investigated the ability of OMVs to stimulate innate and adaptive immunity and to function as a stand-alone adjuvant. We show that OMVs are more potent than heat-inactivated and live-attenuated bacteria in driving dendritic cell activation in vitro and in vivo. Mice immunized with OMVs admixed with heterologous peptides generated peptide-specific CD4 and CD8 T cells responses. Notably, OMV adjuvant induced much greater antibody and B cell responses to co-delivered ovalbumin compared to the responses elicited by the adjuvants alum and CpG DNA. Additionally, pre-existing antibodies raised against the OMVs did not impair OMV adjuvanticity upon repeat immunization. These results indicate that vaccines adjuvanted with OMVs elicit robust cellular and humoral immune responses, supporting further development of OMV adjuvant for use in next-generation vaccines.
Image Deconvolution with Hybrid Reweighted Adaptive Total Variation (HRATV) for Optoacoustic Tomography
Authors: Yang Chen , Jiao Yang , Jian Xiaohua , Cui Yaoyao
Published: January 2021

Optoacoustic tomography (OAT) is a hybrid biomedical imaging modality that usually employs a transducer array to detect laser-generated ultrasonic signals. The reconstructed image suffers low contrast and degraded resolution due to the limited bandwidth and the spatial directivity of the transducer element. Here, we introduce a modified image deconvolution method with a hybrid reweighted adaptive total variation tailored to improve the image quality of OAT. The effectiveness and the parameter dependency of the proposed method are verified on standard test images. The performance of the proposed method in OAT is then characterized on both simulated phantoms and in vivo mice experiments, which demonstrates that the modified deconvolution algorithm is able to restore the sharp edges and fine details in OAT simultaneously. The signal-to-noise ratios (SNRs) of the target structures in mouse liver and brain were improved by 4.90 and 12.69 dB, respectively. We also investigated the feasibility of using Fourier ring correlation (FRC) as an indicator of the image quality to monitor the deconvolution progress in OAT. Based on the experimental results, a practical guide for image deconvolution in OAT was summarized. We anticipate that the proposed method will be a promising post-processing tool to enhance the visualization of micro-structures in OAT.
The Role of Botanical Families in Medicinal Ethnobotany: A Phylogenetic Perspective
Authors: Gras Airy , Hidalgo Oriane , D’Ambrosio Ugo , Parada Montse , Garnatje Teresa , Vallès Joan
Published: January 2021

Studies suggesting that medicinal plants are not chosen at random are becoming more common. The goal of this work is to shed light on the role of botanical families in ethnobotany, depicting in a molecular phylogenetic frame the relationships between families and medicinal uses of vascular plants in several Catalan-speaking territories. The simple quantitative analyses for ailments categories and the construction of families and disorders matrix were carried out in this study. A Bayesian approach was used to estimate the over- and underused families in the medicinal flora. Phylogenetically informed analyses were carried out to identify lineages in which there is an overrepresentation of families in a given category of use, i.e., hot nodes. The ethnobotanicity index, at a specific level, was calculated and also adapted to the family level. Two diversity indices to measure the richness of reported taxa within each family were calculated. A total of 47,630 use reports were analysed. These uses are grouped in 120 botanical families. The ethnobotanicity index for this area is 14.44% and the ethnobotanicity index at the family level is 68.21%. The most-reported families are Lamiaceae and Asteraceae and the most reported troubles are disorders of the digestive and nutritional system. Based on the meta-analytic results, indicating hot nodes of useful plants at the phylogenetic level, specific ethnopharmacological research may be suggested, including a phytochemical approach of particularly interesting taxa.
Whole-Genome DNA Methylation Analysis in Hydrogen Peroxide Overproducing Transgenic Tobacco Resistant to Biotic and Abiotic Stresses
Authors: Villagómez-Aranda Ana L. , García-Ortega Luis F. , Torres-Pacheco Irineo , Guevara-González Ramón G.
Published: January 2021

Epigenetic regulation is a key component of stress responses, acclimatization and adaptation processes in plants. DNA methylation is a stable mark plausible for the inheritance of epigenetic traits, such that it is a potential scheme for plant breeding. However, the effect of modulators of stress responses, as hydrogen peroxide (H2O2), in the methylome status has not been elucidated. A transgenic tobacco model to the CchGLP gene displayed high H2O2 endogen levels correlated with biotic and abiotic stresses resistance. The present study aimed to determine the DNA methylation status changes in the transgenic model to obtain more information about the molecular mechanism involved in resistance phenotypes. The Whole-genome bisulfite sequencing analysis revealed a minimal impact of overall levels and distribution of methylation. A total of 9432 differential methylated sites were identified in distinct genome regions, most of them in CHG context, with a trend to hypomethylation. Of these, 1117 sites corresponded to genes, from which 83 were also differentially expressed in the plants. Several genes were associated with respiration, energy, and calcium signaling. The data obtained highlighted the relevance of the H2O2 in the homeostasis of the system in stress conditions, affecting at methylation level and suggesting an association of the H2O2 in the physiological adaptation to stress functional linkages may be regulated in part by DNA methylation.
Abiotic Stress in Crop Species: Improving Tolerance by Applying Plant Metabolites
Authors: Godoy Francisca , Olivos-Hernández Karina , Stange Claudia , Handford Michael
Published: January 2021

Reductions in crop yields brought about by abiotic stress are expected to increase as climate change, and other factors, generate harsher environmental conditions in regions traditionally used for cultivation. Although breeding and genetically modified and edited organisms have generated many varieties with greater abiotic stress tolerance, their practical use depends on lengthy processes, such as biological cycles and legal aspects. On the other hand, a non-genetic approach to improve crop yield in stress conditions involves the exogenous application of natural compounds, including plant metabolites. In this review, we examine the recent literature related to the application of different natural primary (proline, l-tryptophan, glutathione, and citric acid) and secondary (polyols, ascorbic acid, lipoic acid, glycine betaine, α-tocopherol, and melatonin) plant metabolites in improving tolerance to abiotic stress. We focus on drought, saline, heavy metal, and temperature as environmental parameters that are forecast to become more extreme or frequent as the climate continues to alter. The benefits of such applications are often evaluated by measuring their effects on metabolic, biochemical, and morphological parameters in a variety of crop plants, which usually result in improved yields when applied in greenhouse conditions or in the field. As this strategy has proven to be an effective way to raise plant tolerance to abiotic stress, we also discuss the prospect of its widespread implementation in the short term.
Correction: Khorobrykh, A. Hydrogen Peroxide and Superoxide Anion Radical Photoproduction in PSII Preparations at Various Modifications of the Water-Oxidizing Complex. Plants 2019, 8, 329
Authors: Khorobrykh Andrey
Published: January 2021

In the original article, there was a mistake in the legend for ** Figure 5 ** [...]
Genome-Wide Association Study of Natural Variation in Arabidopsis Exposed to Acid Mine Drainage Toxicity and Validation of Associated Genes with Reverse Genetics
Authors: Ghimire Bandana , Saminathan Thangasamy , Bodunrin Abiodun , Abburi Venkata Lakshmi , Kshetry Arjun Ojha , Shinde Suhas , Nimmakayala Padma , Reddy Umesh K.
Published: January 2021

Acid mine drainage (AMD) is a huge environmental problem in mountain-top mining regions worldwide, including the Appalachian Mountains in the United States. This study applied a genome-wide association study (GWAS) to uncover genomic loci in Arabidopsis associated with tolerance to AMD toxicity. We characterized five major root phenotypes—cumulative root length, average root diameter, root surface area, root volume, and primary root length—in 180 Arabidopsis accessions in response to AMD-supplemented growth medium. GWAS of natural variation in the panel revealed genes associated with tolerance to an acidic environment. Most of these genes were transcription factors, anion/cation transporters, metal transporters, and unknown proteins. Two T-DNA insertion mutants, At1g63005 (miR399b) and At2g05635 (DEAD helicase RAD3), showed enhanced acidity tolerance. Our GWAS and the reverse genetic approach revealed genes involved in conferring tolerance to coal AMD. Our results indicated that proton resistance in hydroponic conditions could be an important index to improve plant growth in acidic soil, at least in acid-sensitive plant species.
Antibacterial Synergism of Electrospun Nanofiber Mats Functioned with Silver Nanoparticles and Pulsed Electromagnetic Waves
Authors: El-kaliuoby Mai I. , Khalil Alaa M. , El-Khatib Ahmed M. , Shehata Nader
Published: January 2021

The over-reliance on antibiotics and their enormous misuse has led to warnings of a future without effective medicines and so, the need for alternatives to antibiotics has become a must. Non-traditional antibacterial treatment was performed by using an aray of nanocomposites synergised with exposure to electromagnetic waves. In this manuscript, electrospun poly(vinyl alcohol) (PVA) nanofiber mats embedded with silver nanoparticles (Ag NPs) were synthesized. The nanocomposites were characterized by Transmission Electron Microscope (TEM), Scanning Electron Microscope (SEM), Current-Voltage (I-V) curves, and Thermogravimetric analysis (TGA) along with analysis of antibacterial impact against E. coli and S. aureus bacteria, studied by bacterial growing analysis, growth kinetics, and cellular cytotoxicity. The results indicated a spherical grain shape of silver of average size 20 nm and nanofibers’ mean diameter of less than 100 nm. The nanocomposite mats showed good exposure to bacteria and the ability to sustain release of silver for a relatively long time. Moreover, the applied electromagnetic waves (EMWs) were shown to be a synergistic co-factor in killing bacteria even at low concentrations of Ag NPs. This caused pronounced alterations of the bacterial preserved packing of the cell membrane. Thereby, the treatment with nanocomposite mats under EM wave exposure elucidated maximum inhibition for both bacterial strains. It was concluded that the functioning of nanofiber with silver nanoparticles and exposure to electromagnetic waves improved the antibacterial impact compared to each one alone.
Bioactivity Comparison of Electrospun PCL Mats and Liver Extracellular Matrix as Scaffolds for HepG2 Cells
Authors: Slivac Igor , Zdraveva Emilija , Ivančić Fran , Žunar Bojan , Holjevac Grgurić Tamara , Gaurina Srček Višnja , Svetec Ivan-Krešimir , Dolenec Tamara , Bajsić Emi Govorčin , Tominac Trcin Mirna , Mijović Budimir
Published: January 2021

Cells grown on bioactive matrices have immensely advanced many aspects of biomedical research related to drug delivery and tissue engineering. Our main objective was to perform simple evaluation of the structural and biotic qualities of cell scaffolds made of affordable biomaterials for liver cell line (HepG2) cultivation in vitro. In this work the electrospun matrix made of synthetic polyester poly(ε-caprolactone) (PCL) was compared with the natural protein-based extracellular matrix isolated from porcine liver (ECM). Mechanical and structural analysis showed that ECM was about 12 times less resistant to tensile stress while it had significantly larger pore size and twice smaller water contact angle than PCL. Bioactivity assessment included comparison of cell growth and transfection efficiency on cell-seeded scaffolds. Despite the differences in composition and structure between the two respective matrices, the rate of cell spreading and the percentage of transfected cells on both scaffolds were fairly comparable. These results suggest that in an attempt to produce simple, cell carrying structures that adequately simulate the natural scaffold, one can rely on PCL electrospun mats.
Migration and Degradation in Composting Environment of Active Polylactic Acid Bilayer Nanocomposites Films: Combined Role of Umbelliferone, Lignin and Cellulose Nanostructures
Authors: Iglesias-Montes Magdalena L. , Luzi Francesca , Dominici Franco , Torre Luigi , Manfredi Liliana B. , Cyras Viviana P. , Puglia Debora
Published: January 2021

This study was dedicated to the functional characterization of innovative poly(lactic acid) (PLA)-based bilayer films containing lignocellulosic nanostructures (cellulose nanocrystals (CNCs) or lignin nanoparticles (LNPs)) and umbelliferone (UMB) as active ingredients (AIs), prepared to be used as active food packaging. Materials proved to have active properties associated with the antioxidant action of UMB and LNPs, as the combination of both ingredients in the bilayer formulations produced a positive synergic effect inducing the highest antioxidant capacity. The results of overall migration for the PLA bilayer systems combining CNCs or LNPs and UMB revealed that none of these samples exceeded the overall migration limit required by the current normative for food packaging materials in both non-polar and polar simulants. Finally, all the hydrophobic monolayer and bilayer films were completely disintegrated in composting conditions in less than 18 days of incubation, providing a good insight on the potential use of these materials for application as active and compostable food packaging.
Densification: A Route towards Enhanced Thermal Conductivity of Epoxy Composites
Authors: Moradi Sasan , Román Frida , Calventus Yolanda , Hutchinson John M.
Published: January 2021

When an amorphous polymer is cooled under pressure from above its glass transition temperature to room temperature, and then the pressure is released, this results in a densified state of the glass. This procedure applied to an epoxy composite system filled with boron nitride (BN) particles has been shown to increase the density of the composite, reduce its enthalpy, and, most importantly, significantly enhance its thermal conductivity. An epoxy-BN composite with 58 wt% BN platelets of average size 30 µm has been densified by curing under pressures of up to 2.0 MPa and then cooling the cured sample to room temperature before releasing the pressure. It is found that the thermal conductivity is increased from approximately 3 W/mK for a sample cured at ambient pressure to approximately 7 W/mK; in parallel, the density increases from 1.55 to 1.72 ± 0.01 g/cm3. This densification process is much more effective in enhancing the thermal conductivity than is either simply applying pressure to consolidate the epoxy composite mixture before curing or applying pressure during cure but then removing the pressure before cooling to room temperature; this last procedure results in a thermal conductivity of approximately 5 W/mK. Furthermore, it has been shown that the densification and corresponding effect on the thermal conductivity is reversible; it can be removed by heating above the glass transition temperature and then cooling without pressure and can be reinstated by again heating above the glass transition temperature and then cooling under pressure. This implies that a densified state and an enhanced thermal conductivity can be induced even in a composite prepared without the use of pressure.
Pilot-Scale Production of Chito-Oligosaccharides Using an Innovative Recombinant Chitosanase Preparation Approach
Authors: Cheng Chih-Yu , Tsai Chia-Huang , Liou Pei-Jyun , Wang Chi-Hang
Published: January 2021

For pilot-scale production of chito-oligosaccharides, it must be cost-effective to prepare designable recombinant chitosanase. Herein, an efficient method for preparing recombinant Bacillus chitosanase from Escherichia coli by elimination of undesirable substances as a precipitate is proposed. After an optimized culture with IPTG (Isopropyl β-d-1-thiogalactopyranoside) induction, the harvested cells were resuspended, disrupted by sonication, divided by selective precipitation, and stored using the same solution conditions. Several factors involved in these procedures, including ion types, ionic concentration, pH, and bacterial cell density, were examined. The optimal conditions were inferred to be pH = 4.5, 300 mM sodium dihydrogen phosphate, and cell density below 1011 cells/mL. Finally, recombinant chitosanase was purified to >70% homogeneity with an activity recovery and enzyme yield of 90% and 106 mg/L, respectively. When 10 L of 5% chitosan was hydrolyzed with 2500 units of chitosanase at ambient temperature for 72 h, hydrolyzed products having molar masses of 833 ± 222 g/mol with multiple degrees of polymerization (chito-dimer to tetramer) were obtained. This work provided an economical and eco-friendly preparation of recombinant chitosanase to scale up the hydrolysis of chitosan towards tailored oligosaccharides in the near future.
Influence of Drug Incorporation on the Physico-Chemical Properties of Poly(l-Lactide) Implant Coating Matrices—A Systematic Study
Authors: Arbeiter Daniela , Reske Thomas , Teske Michael , Bajer Dalibor , Senz Volkmar , Schmitz Klaus-Peter , Grabow Niels , Oschatz Stefan
Published: January 2021

Local drug delivery has become indispensable in biomedical engineering with stents being ideal carrier platforms. While local drug release is superior to systemic administration in many fields, the incorporation of drugs into polymers may influence the physico-chemical properties of said matrix. This is of particular relevance as minimally invasive implantation is frequently accompanied by mechanical stresses on the implant and coating. Thus, drug incorporation into polymers may result in a susceptibility to potentially life-threatening implant failure. We investigated spray-coated poly-l-lactide (PLLA)/drug blends using thermal measurements (DSC) and tensile tests to determine the influence of selected drugs, namely sirolimus, paclitaxel, dexamethasone, and cyclosporine A, on the physico-chemical properties of the polymer. For all drugs and PLLA/drug ratios, an increase in tensile strength was observed. As for sirolimus and dexamethasone, PLLA/drug mixed phase systems were identified by shifted drug melting peaks at 200 °C and 240 °C, respectively, whereas paclitaxel and dexamethasone led to cold crystallization. Cyclosporine A did not affect matrix thermal properties. Altogether, our data provide a contribution towards an understanding of the complex interaction between PLLA and different drugs. Our results hold implications regarding the necessity of target-oriented thermal treatment to ensure the shelf life and performance of stent coatings.
On the Mutual Relationships between Molecular Probe Mobility and Free Volume and Polymer Dynamics in Organic Glass Formers: cis-1,4-poly(isoprene)
Authors: Švajdlenková Helena , Šauša Ondrej , Adichtchev Sergey V. , Surovtsev Nikolay V. , Novikov Vladimir N. , Bartoš Josef
Published: January 2021

We report on the reorientation dynamics of small spin probe 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO) in cis-1,4-poly(isoprene) (cis-1,4-PIP10k) from electron spin resonance (ESR) and the free volume of cis-1,4-PIP10k from positron annihilation lifetime spectroscopy (PALS) in relation to the high-frequency relaxations of cis-1,4-PIP10k using light scattering (LS) as well as to the slow and fast processes from broadband dielectric spectroscopy (BDS) and neutron scattering (NS). The hyperfine coupling constant, 2Azz′(T), and the correlation times, τc(T), of cis-1,4-PIP10k/TEMPO system as a function of temperature exhibit several regions of the distinct spin probe TEMPO dynamics over a wide temperature range from 100 K up to 350 K. The characteristic ESR temperatures of changes in the spin probe dynamics in cis-1,4-PIP10k/TEMPO system are closely related to the characteristic PALS ones reflecting changes in the free volume expansion from PALS measurement. Finally, the time scales of the slow and fast dynamics of TEMPO in cis-1,4-PIP10k are compared with all of the six known slow and fast relaxation modes from BDS, LS and NS techniques with the aim to discuss the controlling factors of the spin probe reorientation mobility in polymer, oligomer and small molecular organic glass-formers.
Recycling of Carbon Fiber Reinforced Composite Polymers—Review—Part 1: Volume of Production, Recycling Technologies, Legislative Aspects
Authors: Bledzki Andrzej K. , Seidlitz Holger , Goracy Krzysztof , Urbaniak Magdalena , Rösch Janina J.
Published: January 2021

The paper presents the current volume of international production and global markets of carbon fiber reinforced polymer composites, also regarding the potential development trends. Examples were provided on how to effectively recycle carbon fiber reinforced polymer composites. Legally binding legislation in the EU on polymer composite recycling was given.
New Pure Organic and Peroxide-Free Redox Initiating Systems for Polymerization in Mild Conditions
Authors: Arar Ahmad , Wisson Lilian , Lalevée Jacques
Published: January 2021

Redox initiating systems (RISs) are highly worthwhile for polymerization in mild conditions (at room temperature—RT) without external thermal or light activation. With high performance redox initiating systems RIS, the free radical polymerization FRP can even be carried out under air and without inhibitors/stabilizers removal from the monomers/resins. However, efficient RISs are still based on peroxides or metal complexes. In this work, a pure organic and peroxide-free RIS is presented based on the interaction of a well-selected triarylamine derivative (T4epa) with iodonium salt used as reducing and oxidizing agents, respectively. The redox polymerization (Redox FRP) was followed through pyrometry and thermal imaging experiments. Remarkably, a full control of the work time as well as a high reactivity is observed for mild conditions.
Excellent Fire Retardant Properties of CNF/VMT Based LBL Coatings Deposited on Polypropylene and Wood-Ply
Authors: Rehman Zeeshan Ur , Niaz Atif Khan , Song Jung-Il , Koo Bon Heun
Published: January 2021

In this report, layer by layer (LBL) fire retardant coatings were produced on wood ply and Polypropylene Homopolymer/Flax fiber composites. FE-SEM and EDAX analysis was carried out to analyze the surface morphology, thickness, growth rate and elemental composition of the samples. Coatings with a high degree of uniformity were formed on Polypropylene composite (PP/flax), while coatings with highest thickness were obtained on wood ply (wood). FTIR and Raman spectroscopy were further used for the molecular identifications of the coatings, which confirmed the maximum deposition of the solution components on the wood substrate. A physiochemical analysis and model was proposed to explain the forces of adhesion between the substrate and solution molecules. Fire protection and thermal properties were studied using TGA and UL-94 tests. It was explored, that the degradation of the coated substrates was highly protected by the coatings as follows: wood > PP/flax > PP. From the UL-94 test, it was further discovered that more than 83% of the coated wood substrate was protected from burning, compared to the 0% of the uncoated substrate. The flammability resistance of the samples was ranked as wood > PP/flax > PP.
Average Shear Rates in the Screw Elements of a Corotating Twin-Screw Extruder
Authors: Vergnes Bruno
Published: January 2021

The rapid estimation of the average shear rate encountered by the material as it flows along the screw elements of a corotating twin-screw extruder is a key point for many applications. In this paper, two methods of evaluation are presented that allow the calculation of the average shear rate as a function of the screw geometry, feed rate, and screw speed. A comparison is made between the approximate and exact methods. It is shown that it is crucial to take into account the shear component due to the pressure flow, especially in the left-handed screw elements.
3D Printing of a Self-Healing Thermoplastic Polyurethane through FDM: From Polymer Slab to Mechanical Assessment
Authors: Ritzen Linda , Montano Vincenzo , Garcia Santiago J.
Published: January 2021

The use of self-healing (SH) polymers to make 3D-printed polymeric parts offers the potential to increase the quality of 3D-printed parts and to increase their durability and damage tolerance due to their (on-demand) dynamic nature. Nevertheless, 3D-printing of such dynamic polymers is not a straightforward process due to their polymer architecture and rheological complexity and the limited quantities produced at lab-scale. This limits the exploration of the full potential of self-healing polymers. In this paper, we present the complete process for fused deposition modelling of a room temperature self-healing polyurethane. Starting from the synthesis and polymer slab manufacturing, we processed the polymer into a continuous filament and 3D printed parts. For the characterization of the 3D printed parts, we used a compression cut test, which proved useful when limited amount of material is available. The test was able to quasi-quantitatively assess both bulk and 3D printed samples and their self-healing behavior. The mechanical and healing behavior of the 3D printed self-healing polyurethane was highly similar to that of the bulk SH polymer. This indicates that the self-healing property of the polymer was retained even after multiple processing steps and printing. Compared to a commercial 3D-printing thermoplastic polyurethane, the self-healing polymer displayed a smaller mechanical dependency on the printing conditions with the added value of healing cuts at room temperature.
Solvent-Free Ultrasonic Dispersion of Nanofillers in Epoxy Matrix
Authors: Zanghellini Benjamin , Knaack Patrick , Schörpf Sebastian , Semlitsch Karl-Heinz , Lichtenegger Helga C. , Praher Bernhard , Omastova Maria , Rennhofer Harald
Published: January 2021

Dispersion of carbon nanotubes and carbon nanofibers is a crucial processing step in the production of polymer-based nanocomposites and poses a great challenge due to the tendency of nanofillers to agglomerate. One of the most effective methods for dispersion is the use of a three-roll mill, which is a well-established method and results in agglomerates below 5 µm. Nevertheless, this process is time-consuming and thus a limiting factor for industrial applications. Our aim was to establish an easy and efficient ultrasonic dispersion process, characterize the dispersion parameters, and compare both methods, ultrasonication and the three-roll mill. We applied rheological tests and analyzed the agglomerate sizes by an image fit of the microscopy images. All these analyses combined deliver a valuable set of information about the dispersion’s quality and, therefore, allows the improvement and further adaptation of the dispersion process.
Facile Synthesis of Sprayed CNTs Layer-Embedded Stretchable Sensors with Controllable Sensitivity
Authors: Khalid Hammad R. , Choudhry Iqra , Jang Daeik , Abbas Nadir , Haider M. Salman , Lee H. K.
Published: January 2021

Flexible electronic devices have gained significant interest due to their different potential applications. Herein, we report highly flexible, stretchable, and sensitive sensors made of sprayed CNT layer, sandwiched between two polymer layers. A facile fabrication process was employed in which the CNT solution was directly sprayed onto a patterned bottom polymer layer, above which a second polymer layer was casted to get a sandwiched composite structure. Varying amounts of CNT solution (i.e., 10, 25, 40, 70, and 100 mL) were sprayed to get conductive CNT layers of different thicknesses/densities. The physical characteristics of the conductive CNT layers were studied through SEM and optical images. The starting electrical resistance values (without strain) as well as the changes in electrical resistance against human body motions were monitored. The synthesized samples exhibited good response against finger and wrist bending. The conductivity of the samples increased with increase of CNT solution volume while the sensitivity followed the inverse relation, suggesting that the sensors with controlled sensitivity could be fabricated for targeted strain ranges using the proposed method.