bluuebunny/crossref_metadata_2025 huggingface dataset:
Diacidic Motifs in the Carboxyl Terminus Are Required for ER Exit and Translocation to the Plasma Membrane of NKCC2
Authors: Bakhos-Douaihy Dalal , Seaayfan Elie , Frachon Nadia , Demaretz Sylvie , Kömhoff Martin , Laghmani Kamel
Published: October 2022

Mutations in the apical Na-K-2Cl co-transporter, NKCC2, cause type I Bartter syndrome (BS1), a life-threatening kidney disease. We have previously demonstrated that the BS1 variant Y998X, which deprives NKCC2 from its highly conserved dileucine-like motifs, compromises co-transporter surface delivery through ER retention mechanisms. However, whether these hydrophobic motifs are sufficient for anterograde trafficking of NKCC2 remains to be determined. Interestingly, sequence analysis of NKCC2 C-terminus revealed the presence of consensus di-acidic (D/E-X-D/E) motifs, 949EEE951 and 1019DAELE1023, located upstream and downstream of BS1 mutation Y998X, respectively. Di-acidic codes are involved in ER export of proteins through interaction with COPII budding machinery. Importantly, whereas mutating 949EEE951 motif to 949AEA951 had no effect on NKCC2 processing, mutating 1019DAE1021 to 1019AAA1021 heavily impaired complex-glycosylation and cell surface expression of the cotransporter in HEK293 and OKP cells. Most importantly, triple mutation of D, E and E residues of 1019DAELE1023 to 1019AAALA1023 almost completely abolished NKCC2 complex-glycosylation, suggesting that this mutant failed to exit the ER. Cycloheximide chase analysis demonstrated that the absence of the terminally glycosylated form of 1019AAALA1023 was caused by defects in NKCC2 maturation. Accordingly, co-immunolocalization experiments revealed that 1019AAALA1023 was trapped in the ER. Finally, overexpression of a dominant negative mutant of Sar1-GTPase abolished NKCC2 maturation and cell surface expression, clearly indicating that NKCC2 export from the ER is COPII-dependent. Hence, our data indicate that in addition to the di-leucine like motifs, NKCC2 uses di-acidic exit codes for export from the ER through the COPII-dependent pathway. We propose that any naturally occurring mutation of NKCC2 interfering with this pathway could form the molecular basis of BS1.
Erythropoiesis and Malaria, a Multifaceted Interplay
Authors: Dumarchey Aurélie , Lavazec Catherine , Verdier Frédérique
Published: October 2022

One of the major pathophysiologies of malaria is the development of anemia. Although hemolysis and splenic clearance are well described as causes of malarial anemia, abnormal erythropoiesis has been observed in malaria patients and may contribute significantly to anemia. The interaction between inadequate erythropoiesis and Plasmodium parasite infection, which partly occurs in the bone marrow, has been poorly investigated to date. However, recent findings may provide new insights. This review outlines clinical and experimental studies describing different aspects of ineffective erythropoiesis and dyserythropoiesis observed in malaria patients and in animal or in vitro models. We also highlight the various human and parasite factors leading to erythropoiesis disorders and discuss the impact that Plasmodium parasites may have on the suppression of erythropoiesis.
iTRAQ Proteomics Identified the Potential Biomarkers of Coronary Artery Lesion in Kawasaki Disease and In Vitro Studies Demonstrated That S100A4 Treatment Made HCAECs More Susceptible to Neutrophil Infiltration
Authors: Weng Ken-Pen , Chien Kuang-Jen , Huang Shih-Hui , Huang Lien-Hung , Lin Pei-Hsien , Lin Yuyu , Chang Wei-Hsiang , Chen Chun-Yu , Li Sung-Chou
Published: October 2022

Coronary artery lesions (CAL) are a major complication of Kawasaki disease (KD). The early prediction of CAL enables the medical personnel to apply adequate medical intervention. We collected the serum samples from the KD patients with CAL (n = 32) and those without CAL (n = 31), followed by a global screening with isobaric tagging for relative and absolute quantification (iTRAQ) technology and specific validation with an enzyme-linked immunosorbent assay (ELISA). iTRAQ identified 846 proteins in total in the serum samples, and four candidate proteins related to CAL were selected for ELISA validation as follows: Protein S100-A4 (S100A4), Catalase (CAT), Folate receptor gamma (FOLR3), and Galectin 10 (CLC). ELISA validation showed that the S100A4 level was significantly higher in KD patients with CAL than in those without CAL (225.2 ± 209.5 vs. 143.3 ± 83 pg/mL, p < 0.05). In addition, KD patients with CAL had a significantly lower CAT level than those without CAL (1.6 ± 1.5 vs. 2.7 ± 2.3 ng/mL, p < 0.05). Next, we found that S100A4 treatment on human coronary artery endothelial cells (HCAECs) reduced the abundance of cell junction proteins, which promoted the migration of HCAECs. Further assays also demonstrated that S100A4 treatment enhanced the permeability of the endothelial layer. These results concluded that S100A4 treatment resulted in an incompact endothelial layer and made HCAECs more susceptible to in vitro neutrophil infiltration. In addition, both upregulated S100A4 and downregulated CAT increased the risk of CAL in KD. Further in vitro study implied that S100A4 could be a potential therapeutic target for CAL in KD.
Platelets and Their Role in Hemostasis and Thrombosis—From Physiology to Pathophysiology and Therapeutic Implications
Authors: Scridon Alina
Published: October 2022

Hemostasis is a physiological process critical for survival. Meanwhile, thrombosis is amongst the leading causes of death worldwide, making antithrombotic therapy one of the most crucial aspects of modern medicine. Although antithrombotic therapy has progressed tremendously over the years, it remains far from ideal, and this is mainly due to the incomplete understanding of the exceptionally complex structural and functional properties of platelets. However, advances in biochemistry, molecular biology, and the advent of ‘omics’ continue to provide crucial information for our understanding of the complex structure and function of platelets, their interactions with the coagulation system, and their role in hemostasis and thrombosis. In this review, we provide a comprehensive view of the complex role that platelets play in hemostasis and thrombosis, and we discuss the major clinical implications of these fundamental blood components, with a focus on hemostatic platelet-related disorders and existing and emerging antithrombotic therapies. We also emphasize a number of questions that remain to be answered, and we identify hotspots for future research.
Rhus Coriaria L. Extract: Antioxidant Effect and Modulation of Bioenergetic Capacity in Fibroblasts from Parkinson’s Disease Patients and THP-1 Macrophages
Authors: Isgrò Camilla , Spagnuolo Ludovica , Pannucci Elisa , Mondello Luigi , Santi Luca , Dugo Laura , Sardanelli Anna Maria
Published: October 2022

Sumac, Rhus coriaria L., is a Mediterranean plant showing several useful properties, such as antioxidant and neuroprotective effects. Currently, there is no evidence about its possible neuroprotective action in Parkinson’s disease (PD). We hypothesized that sumac could modulate mitochondrial functionality in fibroblasts of familial early-onset PD patients showing PARK2 mutations. Sumac extract volatile profile, polyphenolic content and antioxidant activity have been previously characterized. We evaluated ROS and ATP levels on sumac-treated patients’ and healthy control fibroblasts. In PD fibroblasts, all treatments were effective in reducing H2O2 levels, while patients’ ATP content was modulated differently, probably due to the varying mutations in the PARK2 gene found in individual patients which are also involved in different mitochondrial phenotypes. We also investigated the effect of sumac extract on THP-1-differentiated macrophages, which show different embryogenic origin with respect to fibroblasts. In THP-1 macrophages, sumac treatment determined a reduction in H2O2 levels and an increase in the mitochondrial ATP content in M1, assuming that sumac could polarize the M1 to M2 phenotype, as demonstrated with other food-derived compounds rich in polyphenols. In conclusion, Rhus coriaria L. extracts could represent a potential nutraceutical approach to PD.
Effects and Mechanism of Enhanced UV-B Radiation on the Flag Leaf Angle of Rice
Authors: Ling Chengting , Wang Xiupin , Li Zuran , He Yongmei , Li Yuan
Published: October 2022

Leaf angle is an influential agricultural trait that influences rice (Oryza sativa L.) plant type and yield, which results from the leaf bending from the vertical axis to the abaxial axis. UV-B radiation affects plant morphology, but the effects of varying UV-B intensities on rice flag leaves and the underlying molecular, cellular, and physiological mechanisms remain unknown. This experiment aims to examine the effect of natural light and field-enhanced UV-B radiation (2.5, 5.0, 7.5 kJ·m−2) on the leaf angle of the traditional rice variety Baijiaolaojing on Yuanyang terraces. In comparison with natural light, the content of brassinolide and gibberellin in rice flag leaves increased by 29.94% and 60.1%, respectively. The auxin content decreased by 17.3%. Compared with the natural light treatment, the cellulose content in the pulvini was reduced by 13.8% and hemicellulose content by 25.7% under 7.5 kJ·m−2 radiation intensity. The thick-walled cell area and vascular bundle area of the leaf pulvini decreased with increasing radiation intensity, and the growth of mechanical tissue in the rice leaf pulvini was inhibited. The flag leaf angle of rice was greatest at 7.5 kJ·m−2 radiation intensity, with an increase of 50.2%. There are two pathways by which the angle of rice flag leaves is controlled under high-intensity UV-B radiation. The leaf angle regulation genes OsBUL1, OsGSR1, and OsARF19 control hormone levels, whereas the ILA1 gene controls fiber levels. Therefore, as cellulose, hemicellulose, sclerenchyma, and vascular bundles weaken the mechanical support of the pulvini, the angle of the flag leaf increases.
Ab Initio Study of Carrier Mobility, Thermodynamic and Thermoelectric Properties of Kesterite Cu2ZnGeS4
Authors: El Hamdaoui Jawad El , Kria Mohamed , Lakaal Kamal , El-Yadri Mohamed , Feddi El Mustapha , Pedraja Rejas Liliana Pedraja , Pérez Laura M. , Díaz Pablo , Mora-Ramos Miguel E. , Laroze David
Published: October 2022

The kesterite Cu2ZnGeS4 (CZGS) has recently gained significant interest in the scientific community. In this work, we investigated the thermodynamic and thermoelectric properties of CZGS by employing the first-principals calculation in association with the quasi-harmonic approximation, Boltzmann transport theory, deformation potential theory, and slack model. We obtained a bandgap of 2.05 eV and high carrier mobility. We found that CZGS exhibits adequate thermoelectric properties as a promising material for thermoelectric applications. The calculated Seebeck coefficient at room temperature is 149 µV·K−1. We also determined the thermal and electrical conductivity, the power factor, and the figure of merit. In addition, the thermodynamic properties such as Debye temperature, entropy, and constant volume heat capacity are estimated. According to our results, it is concluded that the Slack model fails to provide correct values for lattice thermal conductivity in this material.
Characterization of Abscisic Acid and Ethylene in Regulating the White Blush in Fresh-Cut Carrots
Authors: Chen Chichang , Wang Ye , Xu Li , Wang Siwen , Chen Xiangning , Shen Yuanyue
Published: October 2022

The surface of fresh-cut carrots is apt to white blush, however the physiological and molecular mechanism for this process is not yet fully understood. In this study, exogenous abscisic acid (ABA) and ethylene separately promoted and inhibited the white-blush formation after three days after treatment, respectively. Metabolome analysis found that white-blush components mainly consist of p-hydroxyphenyl lignin and guaiacyl lignin. Transcriptome analysis found an increase in the whiteness values was consistent with the higher expression of genes encoding O-methyltransferase, trans-anol O-methyltransferase, bergaptol O-methyltransferase, caffeic acid 3-O-methyltransferase, phenylalanine ammonia-lyase, and ferulate-5-hydroxylase, together with the lower expression of genes encoding cinnamic acid 4-hydroxylase caffeoyl-CoA O-methyltransferase and 5-O-(4-coumaroyl)-D-quinate 3′-monooxygenase. In conclusion, ABA plays an important role in lignin biosynthesis essential to the formation of white blush in fresh-cut carrots. This is the first report that uncovers the physiological and molecular causes of white blush in fresh-cut carrots, providing a basis for white-blush control in fresh-cut carrots.
Does Vitamin D Work Synergistically with Anti-Asthmatic Drugs in Airway Remodeling?
Authors: Sobczak Marharyta , Pawliczak Rafał
Published: October 2022

Vitamin D is commonly known for its properties of airway remodeling inhibition. Due to this, we decided to analyze the action of calcitriol with anti-asthmatic drugs in airway remodeling. The HFL1 cell line was treated with calcitriol, beclomethasone 17-propionate, montelukast sodium, LTD4 and TGF-β in different combinations. Real-time PCR was used to analyzed the expression of ACTA2, CDH-1, Vimentin, ADAM33, MMP-9 and CysLTR1 on the mRNA level, whereas Western blot was used to analyze gene expression on the protein level. One-way analysis variants, the Kruskal-Wallis test, Student’s t-test or Welch’s t-test were used for statistical analysis. Concerning the results, pre-treatment with calcitriol increased the inhibitory effect of beclomethasone 17-propionate and montelukast sodium on the expression of ACTA2 (p = 0.0072), Vimentin (p = 0.0002) and CysLTR1 (p = 0.0204), and 1,25(OH)2D3 had an influence on the effects of beclomethasone 17-propionate and montelukast sodium and of CDH1 expression (p = 0.0076). On the protein level, pre-treatment with calcitriol with beclomethasone 17-propionate and montelukast sodium treatment decreased ACTA2 expression in comparison to the LT (LTD4 and TGF-β) control group (p = 0.0191). Hence, our study not only confirms that vitamin D may inhibit airway remodeling, but also shows that vitamin D has a synergistic effect with anti-asthmatic drugs.
Defining the Functional Interactome of Spliceosome-Associated G-Patch Protein Gpl1 in the Fission Yeast Schizosaccharomyces pombe
Authors: Selicky Tomas , Jurcik Matus , Mikolaskova Barbora , Pitelova Alexandra , Mayerova Nina , Kretova Miroslava , Osadska Michaela , Jurcik Jan , Holic Roman , Kohutova Lenka , Bellova Jana , Benko Zsigmond , Gregan Juraj , Bagelova Polakova Silvia , Barath Peter , Cipak Lubos , Cipakova Ingrid
Published: October 2022

Pre-mRNA splicing plays a fundamental role in securing protein diversity by generating multiple transcript isoforms from a single gene. Recently, it has been shown that specific G-patch domain-containing proteins are critical cofactors involved in the regulation of splicing processes. In this study, using the knock-out strategy, affinity purification and the yeast-two-hybrid assay, we demonstrated that the spliceosome-associated G-patch protein Gpl1 of the fission yeast S. pombe mediates interactions between putative RNA helicase Gih35 (SPAC20H4.09) and WD repeat protein Wdr83, and ensures their binding to the spliceosome. Furthermore, RT-qPCR analysis of the splicing efficiency of deletion mutants indicated that the absence of any of the components of the Gpl1-Gih35-Wdr83 complex leads to defective splicing of fet5 and pwi1, the reference genes whose unspliced isoforms harboring premature stop codons are targeted for degradation by the nonsense-mediated decay (NMD) pathway. Together, our results shed more light on the functional interactome of G-patch protein Gpl1 and revealed that the Gpl1-Gih35-Wdr83 complex plays an important role in the regulation of pre-mRNA splicing in S. pombe.
MGL S3 Chimeric Enzyme Drives Apoptotic Death of EGFR-Dependent Cancer Cells through ERK Downregulation
Authors: Bondarev Nikolay , Ivanenko Karina , Khabusheva Elmira , Lebedev Timofey , Manukhov Ilya , Prassolov Vladimir
Published: October 2022

Methionine dependence of malignant cells is one of the cancer hallmarks. It is well described that methionine deprivation drives cancer cells death, both in vitro and in vivo. Methionine gamma-lyase (MGL) isolated from different species or obtained by genetic engineering can be used for effective methionine depletion. In this work, we show that MGL S3, a genetically engineered protein comprised of MGL from Clostridium sporogenesis fused to epidermal growth factor (EGF)-like peptide, reduces, in vitro, the number of cancer cells of four different origins—neuroblastoma, lung, breast, and colon cancer. We reveal that MGL S3 is more toxic for neuroblastoma SH-SY5Y and lung cancer H1299 cells compared to MGL tetani, and causes cell death by the induction of apoptosis. In addition, the observed death of cells treated with MGL S3 is accompanied by the prominent downregulation of ERK activity. By the analysis of transcriptomic data of more than 1500 cancer cell lines and patient samples, we show that the high expression of four genes from the methionine metabolism pathway (AHCY, CBS, DNMT3A, and MTAP) is associated with poor prognosis for breast cancer and neuroblastoma patients. Additionally, cells of these origins are characterized by a high correlation between EGFR dependency and DNMT3A/CBS expression. Finally, we demonstrate the ability of MGL S3 to enhance the sensitivity of H1299 cells to EGFR inhibition with gefitinib.
Tuning Immobilized Enzyme Features by Combining Solid-Phase Physicochemical Modification and Mineralization
Authors: Guimarães José R. , Carballares Diego , Rocha-Martin Javier , Tardioli Paulo W. , Fernandez-Lafuente Roberto
Published: October 2022

Lipase B from Candida antarctica (CALB) and lipase from Thermomyces lanuginosus (TLL) were immobilized on octyl agarose. Then, the biocatalysts were chemically modified using glutaraldehyde, trinitrobenzenesulfonic acid or ethylenediamine and carbodiimide, or physically coated with ionic polymers, such as polyethylenimine (PEI) and dextran sulfate. These produced alterations of the enzyme activities have, in most cases, negative effects with some substrates and positive with other ones (e.g., amination of immobilized TLL increases the activity versus p-nitro phenyl butyrate (p-NPB), reduces the activity with R-methyl mandate by half and maintains the activity with S-isomer). The modification with PEI increased the biocatalyst activity 8-fold versus R-methyl mandelate. Enzyme stability was also modified, usually showing an improvement (e.g., the modification of immobilized TLL with PEI or glutaraldehyde enabled to maintain more than 70% of the initial activity, while the unmodified enzyme maintained less than 50%). The immobilized enzymes were also mineralized by using phosphate metals (Zn2+, Co2+, Cu2+, Ni2+ or Mg2+), and this affected also the enzyme activity, specificity (e.g., immobilized TLL increased its activity after zinc mineralization versus triacetin, while decreased its activity versus all the other assayed substrates) and stability (e.g., the same modification increase the residual stability from almost 0 to more than 60%). Depending on the enzyme, a metal could be positively, neutrally or negatively affected for a specific feature. Finally, we analyzed if the chemical modification could, somehow, tune the effects of the mineralization. Effectively, the same mineralization could have very different effects on the same immobilized enzyme if it was previously submitted to different physicochemical modifications. The same mineralization could present different effects on the enzyme activity, specificity or stability, depending on the previous modification performed on the enzyme, showing that these previous enzyme modifications alter the effects of the mineralization on enzyme features. For example, TLL modified with glutaraldehyde and treated with zinc salts increased its activity using R-methyl mandelate, while almost maintaining its activity versus the other unaltered substrates, whereas the aminated TLL maintained its activity with both methyl mandelate isomers, while it decreased with p-NPB and triacetin. TLL was found to be easier to tune than CALB by the strategies used in this paper. In this way, the combination of chemical or physical modifications of enzymes before their mineralization increases the range of modification of features that the immobilized enzyme can experienced, enabling to enlarge the biocatalyst library.
Genome-Wide Characterization of Chrysanthemum indicum Nuclear Factor Y, Subunit C Gene Family Reveals the Roles of CiNF-YCs in Flowering Regulation
Authors: Wang Xueting , Yao Yao , Wen Shiyun , Bin Jing , Tan Qinghua , Lou Jinpeng , Xie Li , Zeng Ruizhen , Guo Herong , Zhang Zhisheng , Wei Qian
Published: October 2022

Nuclear Factor Y, Subunit C (NF-YC) transcription factors are conserved in most plants, and play essential roles in plant growth and development, especially in flowering regulation. Chrysanthemums are important commercial plants, and their market value is strongly impacted by flowering time. Until now, no details regarding the NF-YC family in the Chrysanthemum genus have been available. In this study, five NF-YC genes were cloned from Chrysanthemum indicum. Multiple alignments showed that CiNF-YCs had the highly conserved characteristic regions. Phylogenetic analyses identified a pair of paralogue NF-YC proteins in chrysanthemums. Gene structure and conserved motifs were also analyzed for functional understanding. According to the results of the expression experiments, CiNF-YC1 and CiNF-YC5 were mainly expressed in leaves or flowers, and their expression levels varied greatly from the seedling to flower bud differentiation stage. Arabidopsis overexpressing CiNF-YC1 and CiNF-YC3 showed significantly delayed flowering, accompanied by other morphological alterations. RT-qPCR analysis revealed that genes associated with photoperiod, vernalization, aging, and gibberellin pathways were downregulated in CiNF-YC1-OX lines, relative to the wild type, whereas in CiNF-YC3-OX lines, only SHORT VEGETATIVE PHASE (AtSVP), the key factor in the ambient temperature pathway, was upregulated. Taken together, these findings suggest that CiNF-YC1 and CiNF-YC3 negatively regulate flowering in Arabidopsis via different flowering pathways.
German Word-Final Devoicing in Naturally-Produced and TTS Speech
Authors: Block Aleese , Predeck Kristin , Zellou Georgia
Published: October 2022

This study explores the production and perception of word-final devoicing in German across text-to-speech (from technology used in common voice-AI “smart” speaker devices—specifically, voices from Apple and Amazon) and naturally produced utterances. First, the phonetic realization of word-final devoicing in German across text-to-speech (TTS) and naturally produced word productions was compared. Acoustic analyses reveal that the presence of cues to a word-final voicing contrast varied across speech types. Naturally produced words with phonologically voiced codas contain partial voicing, as well as longer vowels than words with voiceless codas. However, these distinctions are not present in TTS speech. Next, German listeners completed a forced-choice identification task, in which they heard the words and made coda consonant categorizations, in order to examine the intelligibility consequences of the word-final devoicing patterns across speech types. Intended coda identifications are higher for the naturally produced productions than for TTS. Moreover, listeners systematically misidentified voiced codas as voiceless in TTS words. Overall, this study extends previous literature on speech intelligibility at the intersection of speech synthesis and contrast neutralization. TTS voices tend to neutralize salient phonetic cues present in natural speech. Subsequently, listeners are less able to identify phonological distinctions in TTS. We also discuss how investigating which cues are more salient in natural speech can be beneficial in synthetic speech generation to make them more natural and also easier to perceive.
Coding Metamaterial Analysis Based on 1-Bit Conventional and Cuboid Design Structures for Microwave Applications
Authors: Ramachandran Tayaallen , Faruque Mohammad Rashed Iqbal , Islam Mohammad Tariqul , Khandaker Mayeen Uddin , Osman Hamid , Boukhris Imed
Published: October 2022

This study aimed to investigate the compact 1-bit coding metamaterial design with various conventional and cuboid shapes by analysing the bistatic scattering patterns as well as the monostatic radar cross-section for microwave applications. The construction of this metamaterial design depends on binary elements. For example, 1-bit coding metamaterial comprises two kinds of unit cell to mimic both coding particles such as ‘0’ and ‘1’ with 0° and 180° phase responses. This study adopted a 1 mm × 1 mm of epoxy resin fibre (FR-4) substrate material, which possesses a dielectric constant of 4.3 and tangent loss of 0.025, to construct both elements for the 1-bit coding metamaterial. All simulations were performed using the well-known Computer Simulation Technology (CST) software. The elements were selected via a trial-and-error method based on the phase response properties of the designs. On the other hand, the phase response properties from CST software were validated through the comparison of the phase response properties of both elements with the analytical data from HFSS software. Clear closure was obtained from these findings, and it was concluded that the proposed conventional coding metamaterial manifested the lowest RCS values with an increasing number of lattices. However, the cuboid-shaped design with 20 lattices demonstrated an optimised bistatic scattering pattern of −8.49 dBm2. Additionally, the monostatic RCS values were successfully reduced within the 12 to 18 GHz frequency range with −30 to −10 dBm2 values. In short, the introduced designs were suitable for the proposed application field, and this unique phenomenon is described as the novelty of this study.
Research on High-Temperature Evaluation Indexes and Performance of Qingchuan Rock-SBS Composite Modified Asphalt
Authors: Hao Yan , Ye Yali , Zhuang Chuanyi , Hou Fengjian
Published: October 2022

The phenomenon of structural destabilization damage to asphalt pavement is becoming increasingly serious as a result of high temperatures and heavy traffic. Considering the advantages of Qingchuan rock asphalt (QRA) in its durability, high-temperature rutting resistance, and good compatibility with asphalt, it was proposed to compound rock asphalt with SBS to ameliorate the high-temperature performance of asphalt. In this study, DSR and BBR were used to determine the rheological properties of Qingchuan rock-modified asphalt (QRMA) and Qingchuan rock–SBS-modified asphalt (QRA-SBSMA), and the optimum blending amount of rock asphalt was determined based on the PG classification results. Secondly, four different structures of ‘30 mm AC-10 upper layer (70-A, QRMA, SBSMA, QRA-SBSMA) + 50 mm AC-16 lower layer (70-A)’ double-layer composite specimens were prepared. Multiple high-temperature performance evaluation indexes (G*/sinδ, Ds, rutting depth, micro-strain, Fn, modulus) were used to assess the improvement effect of QRA. Finally, using a 1/3 scale accelerated loading testing machine, we simulated high-temperature, water, and high-temperature coupled environments to assess the impact of high temperature and water on the performance of QRMA and QRA-SBSMA, respectively. The findings demonstrated that QRA can increase the PG classification of 70-A and SBSMA as well as its resistance to high-temperature deformation. Multi-index comprehensive evaluation methods were used to consummate the asphalt high-temperature evaluation system. The QRA-SBSMA had the smallest rutting depth and creep rate and the largest dynamic modulus, characterizing its ability to optimally resist high-temperature rutting and deformation.
Branched Chondroitin Sulfate Oligosaccharides Derived from the Sea Cucumber Acaudina molpadioides Stimulate Neurite Outgrowth
Authors: Wang Weili , Mao Hui , Li Sujuan , Zhang Longlong , Yang Lian , Yin Ronghua , Zhao Jinhua
Published: October 2022

Fucosylated chondroitin sulfate (FCS) from the sea cucumber Acaudina molpadioides (FCSAm) is the first one that was reported to be branched by disaccharide GalNAc-(α1,2)-Fuc3S4S (15%) and sulfated Fuc (85%). Here, four size-homogenous fractions, and seven oligosaccharides, were separated from its β-eliminative depolymerized products. Detailed NMR spectroscopic and MS analyses revealed the oligomers as hexa-, hepta-, octa-, and nonasaccharide, which further confirmed the precise structure of native FCSAm: it was composed of the CS-E-like backbone with a full content of sulfation at O-4 and O-6 of GalNAc in the disaccharide repeating unit, and the branches consisting of sulfated fucose (Fuc4S and Fuc2S4S) and heterodisaccharide [GalNAc-(α1,2)-Fuc3S4S]. Pharmacologically, FCSAm and its depolymerized derivatives, including fractions and oligosaccharides, showed potent neurite outgrowth-promoting activity in a chain length-dependent manner. A comparison of analyses among oligosaccharides revealed that the sulfate pattern of the Fuc branches, instead of the heterodisaccharide, could affect the promotion intensity. Fuc2S4S and the saccharide length endowed the neurite outgrowth stimulation activity most.
Integration of the Exogenous Tuning of Thraustochytrid Fermentation and Sulfur Polymerization of Single-Cell Oil for Developing Plant-like Oils
Authors: Gupta Adarsha , Worthington Max J. H. , Chalker Justin M. , Puri Munish
Published: October 2022

In this study, we have demonstrated a bioprocessing approach encompassing the exogenous addition of low-molecular-weight compounds to tune the fatty acid (FA) profile in a novel thraustochytrid strain to produce desirable FAs. Maximum lipid recovery (38%, dry wt. biomass) was obtained at 1% Tween 80 and 0.25 mg/L of Vitamin B12. The transesterified lipid showed palmitic acid (C16, 35.7% TFA), stearic acid (C18, 2.1% TFA), and oleic acid (C18:1, 18.7% TFA) as the main components of total FAs, which are mainly present in plant oils. Strikingly, D-limonene addition in the fermentation medium repressed the production of polyunsaturated fatty acid (PUFAs). Sulfur-polymerization-guided lipid separation revealed the presence of saturated (SFAs, 53% TFA) and monounsaturated fatty acids (MUFAs, 46.6% TFA) in thraustochytrid oil that mimics plant-oil-like FA profiles. This work is industrially valuable and advocates the use of sulfur polymerization for preparation of plant-like oils through tuneable thraustochytrid lipids.
Total Synthesis of Loroxanthin
Authors: Yamano Yumiko , Tanabe Mari , Shimada Atsushi , Wada Akimori
Published: October 2022

The first total synthesis of loroxanthin (1) was accomplished by Horner-Wadsworth-Emmons reaction of C25-apocarotenal 8 having a silyl-protected 19-hydroxy moiety with C15-phosphonate 25 bearing a silyl-protected 3-hydroxy-ε-end group. Preparation of apocarotenal 8 was achieved via Stille coupling reaction of alkenyl iodide 10 with alkenyl stananne 9, whereas phosphonate 25 was prepared through treatment of ally alcohol 23 with triethyl phosphite and ZnI2. The ally alcohol 23 was derived from the known (3R,6R)-3-hydroxy C15-aldehyde 20, which was obtained by direct optical resolution of racemate 20 using a semi-preparative chiral HPLC column.
Chemopreventive Effect on Human Colon Adenocarcinoma Cells of Styrylquinolines: Synthesis, Cytotoxicity, Proapoptotic Effect and Molecular Docking Analysis
Authors: Bedoya-Betancur Vanesa , Correa Elizabeth , Rendón Juan Pablo , Yepes-Pérez Andrés F. , Cardona-Galeano Wilson , Naranjo Tonny W.
Published: October 2022

Seven styrylquinolines were synthesized in this study. Two of these styrylquinolines are new and were elucidated by spectroscopic analysis. The chemopreventive potential of these compounds was evaluated against SW480 human colon adenocarcinoma cells, its metastatic derivative SW620, and normal cells (HaCaT). According to the results, compounds 3a and 3d showed antiproliferative activity in SW480 and SW620 cells, but their effect seemed to be caused by different mechanisms of action. Compound 3a induced apoptosis independent of ROS production, as evidenced by increased levels of caspase 3, and had an immunomodulatory effect, positively regulating the production of different immunological markers in malignant cell lines. In contrast, compound 3d generated a pro-oxidant response and inhibited the growth of cancer cells, probably by another type of cell death other than apoptosis. Molecular docking studies indicated that the most active compound, 3a, could efficiently bind to the proapoptotic human caspases-3 protein, a result that could provide valuable information on the biochemical mechanism for the in vitro cytotoxic response of this compound in SW620 colon carcinoma cell lines. The obtained results suggest that these compounds have chemopreventive potential against CRC, but more studies should be carried out to elucidate the molecular mechanisms of action of each of them in depth.
Understanding the Antilymphoma Activity of Annona macroprophyllata Donn and Its Acyclic Terpenoids: In Vivo, In Vitro, and In Silico Studies
Authors: Ramírez-Santos Jesica , Calzada Fernando , Mendieta-Wejebe Jessica Elena , Ordoñez-Razo Rosa María , Martinez-Casares Rubria Marlen , Valdes Miguel
Published: October 2022

Annona macroprophyllata Donn (A. macroprophyllata) is used in traditional Mexican medicine for the treatment of cancer, diabetes, inflammation, and pain. In this work, we evaluated the antitumor activity of three acyclic terpenoids obtained from A. macroprophyllata to assess their potential as antilymphoma agents. We identified the terpenoids farnesyl acetate (FA), phytol (PT) and geranylgeraniol (Gg) using gas chromatography–mass spectroscopy (GC-MS) and spectroscopic (1H, and 13C NMR) methods applied to petroleum ether extract of leaves from A. macroprophyllata (PEAm). We investigated antitumor potential in Balb/c mice inoculated with U-937 cells by assessing brine shrimp lethality (BSL), and cytotoxic activity in these cells. In addition, to assess the potential toxicity of PEAm, FA, PT and Gg in humans, we tested their acute oral toxicity in mice. Our results showed that the three terpenoids exhibited considerable antilymphoma and cytotoxic activity. In terms of lethality, we determined a median lethal dose (LD50) for thirteen isolated products of PEAm. Gg, PT and AF all exhibited a higher lethality with values of 1.41 ± 0.42, 3.03 ± 0.33 and 5.82 ± 0.58 µg mL−1, respectively. To assess cytotoxic activity against U-937 cells, we calculated the mean cytotoxic concentration (CC50) and found that FA and PT were closer in respect to the control drug methotrexate (MTX, 0.243 ± 0.007 µM). In terms of antilymphoma activity, we found that FA, PT and Gg considerably inhibited lymph node growth, with median effective doses (ED50) of 5.89 ± 0.39, 6.71 ± 0.31 and 7.22 ± 0.51 mg kg−1 in females and 5.09 ± 0.66, 5.83 ± 0.50 and 6.98 ± 0.57mg kg −1 in males, respectively. Regarding acute oral toxicity, we classified all three terpenoids as category IV, indicating a high safety margin for human administration. Finally, in a molecular docking study of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, we found binding of terpenoids to some amino acids of the catalytic site, suggesting an effect upon activity with a resulting decrease in the synthesis of intermediates involved in the prenylation of proteins involved in cancer progression. Our findings suggest that the acyclic terpenoids FA, PT, and Gg may serve as scaffolds for the development of new treatments for non-Hodgkin’s lymphoma.
Alkaloid Derivative (Z)-3β-Ethylamino-Pregn-17(20)-en Inhibits Triple-Negative Breast Cancer Metastasis and Angiogenesis by Targeting HSP90α
Authors: Liu Xin-Yao , Wang Yu-Miao , Zhang Xiang-Yu , Jia Mei-Qi , Duan Hong-Quan , Qin Nan , Chen Ying , Yu Yang , Duan Xiao-Chuan
Published: October 2022

Metastasis is an important cause of cancer-related death. Previous studies in our laboratory found that pregnane alkaloids from Pachysandra terminalis had antimetastatic activity against breast cancer cells. In the current study, we demonstrated that treatment with one of the alkaloid derivatives, (Z)-3β-ethylamino-pregn-17(20)-en (1), led to the downregulation of the HIF-1α/VEGF/VEGFR2 pathway, suppressed the phosphorylation of downstream molecules Akt, mTOR, FAK, and inhibited breast cancer metastasis and angiogenesis both in vitro and in vivo. Furthermore, the antimetastasis and antiangiogenesis effects of 1 treatment (40 mg/kg) were more effective than that of Sorafenib (50 mg/kg). Surface plasmon resonance (SPR) analysis was performed and the result suggested that HSP90α was a direct target of 1. Taken together, our results suggested that compound 1 might represent a candidate antitumor agent for metastatic breast cancer.
Synthesis and Biological Evaluation of PEGylated MWO4 Nanoparticles as Sonodynamic AID Inhibitors in Treating Diffuse Large B-Cell Lymphoma
Authors: Jiao Junna , Qian Zhuang , Wang Yurong , Liu Mei , Fan Liye , Liu Mengqing , Hao Zichen , Jiao Junrong , Lv Zhuangwei
Published: October 2022

Sonodynamic therapy (SDT) triggered by ultrasound (US) has attracted increasing attention owing to its ability to overcome critical limitations, including low tissue-penetration depth and phototoxicity in photodynamic therapy (PDT). Biogenic metal oxide nanoparticles (NPs) have been used as anti-cancer drugs due to their biocompatibility properties with most biological systems. Here, sonosensitizer MWO4-PEG NPs (M = Fe Mn Co Ni) were synthesized as inhibitors to activation-induced cytidine deaminase (AID), thus neutralizing the extensive carcinogenesis of AID in diffuse large B-cell lymphoma (DLBCL). The physiological properties of these nanomaterials were examined using transmission electron microscopy (TEM). The inhibition of NPs to AID was primarily identified by the affinity interaction prediction between reactive oxygen species (ROS) and AID through molecular dynamics and molecular docking technology. The cell apoptosis and ROS generation in US-triggered NPs treated DLBCL cells (with high levels of AID) were also detected to indicate the sonosensitivity and toxicity of MWO4-PEG NPs to DLBCL cells. The anti-lymphoma studies using DLBCL and AID-deficient DLBCL cell lines indicated a concentration-dependent profile. The synthesized MWO4-PEG NPs in this study manifested good sonodynamic inhibitory effects to AID and well treatment for AID-positive hematopoietic cancers.
Anxiolytic-like Effect of Quercetin Possibly through GABA Receptor Interaction Pathway: In Vivo and In Silico Studies
Authors: Islam Md. Shahazul , Hossain Rajib , Ahmed Taukir , Rahaman Md. Mizanur , Al-Khafaji Khattab , Khan Rasel Ahmed , Sarkar Chandan , Bappi Mehedi Hasan , de Andrade Edlane Martins , Araújo Isaac Moura , Coutinho Henrique Douglas Melo , Kowalska Grażyna , Kowalski Radosław , Hanif Muhammad Asif , Islam Muhammad Torequl
Published: October 2022

Scientific evidence suggests that quercetin (QUR) has anxiolytic-like effects in experimental animals. However, the mechanism of action responsible for its anxiolytic-like effects is yet to be discovered. The goal of this research is to assess QUR’s anxiolytic effects in mouse models to explicate the possible mechanism of action. After acute intraperitoneal (i.p.) treatment with QUR at a dose of 50 mg/kg (i.p.), behavioral models of open-field, hole board, swing box, and light–dark tests were performed. QUR was combined with a GABAergic agonist (diazepam) and/or antagonist (flumazenil) group. Furthermore, in silico analysis was also conducted to observe the interaction of QUR and GABA (α5), GABA (β1), and GABA (β2) receptors. In the experimental animal model, QUR had an anxiolytic-like effect. QUR, when combined with diazepam (2 mg/kg, i.p.), drastically potentiated an anxiolytic effect of diazepam. QUR is a more highly competitive ligand for the benzodiazepine recognition site that can displace flumazenil (2.5 mg/kg, i.p.). In all the test models, QUR acted similar to diazepam, with enhanced effects of the standard anxiolytic drug, which were reversed by pre-treatment with flumazenil. QUR showed the best interaction with the GABA (α5) receptor compared to the GABA (β1) and GABA (β2) receptors. In conclusion, QUR may exert an anxiolytic-like effect on mice, probably through the GABA-receptor-interacting pathway.
Synthesis, Structure and Cytotoxic Properties of Copper(II) Complexes of 2-Iminocoumarins Bearing a 1,3,5-Triazine or Benzoxazole/Benzothiazole Moiety
Authors: Makowska Anna , Sączewski Franciszek , Bednarski Patrick J. , Gdaniec Maria , Balewski Łukasz , Warmbier Magdalena , Kornicka Anita
Published: October 2022

A series of copper(II) complexes of 2-imino-2H-chromen-3-yl-1,3,5-triazines 2a-h, 3-(benzoxazol-2-yl)-2H-chromen-2-imines 4a-b, and 3-(benzothiazol-2-yl)-2H-chromen-2-imines 6a-c were obtained by reacting of appropriate 2-iminocoumarin ligands L1a-h, L3a-b, and L5a-c with 3-fold molar excess of copper(II) chloride. The structure of these compounds was confirmed by IR spectroscopy, elemental analysis, and single-crystal X-ray diffraction data (2f, 2g, 2h, and 6c). All the synthesized complexes were screened for their activity against five human cancer cell lines: DAN-G, A-427, LCLC-103H, SISO, and RT-4 by using a crystal violet microtiter plate assay and relationships between structure and in vitro cytotoxic activity are discussed. The coordination of 2-iminocoumarins with copper(II) ions resulted in complexes 2a-h, 4a-b, and 6a-c with significant inhibitory properties toward tested tumor cell lines with IC50 values ranging from 0.04 μM to 15.66 μM. In comparison to the free ligands L1a-h, L3a-b, and L5a-c, the newly prepared Cu(II) complexes often displayed increased activity. In the series of copper(II) complexes of 2-imino-2H-chromen-3-yl-1,3,5-triazines 2a-h the most potent compound 2g contained a 4-phenylpiperazine moiety at position 6 of the 1,3,5-triazine ring and an electron-donating diethylamino group at position 7′ of the 2-iminocoumarin scaffold. Among the Cu(II) complexes of 3-(benzoxazol-2-yl)-2H-chromen-2-imines 4a-b and 3-(benzothiazol-2-yl)-2H-chromen-2-imines 6a-c the most active was benzoxazole-2-iminocoumarin 4b that also possessed a diethylamino group at position 7′ of the 2-iminocoumarin moiety. Moreover, compound 4b was found to be the most prominent agent and displayed the higher potency than cisplatin against tested cell lines.
Review of Functional Aspects of Nanocellulose-Based Pickering Emulsifier for Non-Toxic Application and Its Colloid Stabilization Mechanism
Authors: Teo Shao Hui , Chee Ching Yern , Fahmi Mochamad Zakki , Wibawa Sakti Satya Candra , Lee Hwei Voon
Published: October 2022

In the past few years, the research on particle-stabilized emulsion (Pickering emulsion) has mainly focused on the usage of inorganic particles with well-defined shapes, narrow size distributions, and chemical tunability of the surfaces such as silica, alumina, and clay. However, the presence of incompatibility of some inorganic particles that are non-safe to humans and the ecosystem and their poor sustainability has led to a shift towards the development of materials of biological origin. For this reason, nano-dimensional cellulose (nanocellulose) derived from natural plants is suitable for use as a Pickering material for liquid interface stabilization for various non-toxic product formulations (e.g., the food and beverage, cosmetic, personal care, hygiene, pharmaceutical, and biomedical fields). However, the current understanding of nanocellulose-stabilized Pickering emulsion still lacks consistency in terms of the structural, self-assembly, and physio-chemical properties of nanocellulose towards the stabilization between liquid and oil interfaces. Thus, this review aims to provide a comprehensive study of the behavior of nanocellulose-based particles and their ability as a Pickering functionality to stabilize emulsion droplets. Extensive discussion on the characteristics of nanocelluloses, morphology, and preparation methods that can potentially be applied as Pickering emulsifiers in a different range of emulsions is provided. Nanocellulose’s surface modification for the purpose of altering its characteristics and provoking multifunctional roles for high-grade non-toxic applications is discussed. Subsequently, the water–oil stabilization mechanism and the criteria for effective emulsion stabilization are summarized in this review. Lastly, we discuss the toxicity profile and risk assessment guidelines for the whole life cycle of nanocellulose from the fresh feedstock to the end-life of the product.
Antibacterial Activity of Dental Composite with Ciprofloxacin Loaded Silver Nanoparticles
Authors: Arif Wafa , Rana Nosheen , Saleem Iqra , Tanweer Tahreem , Khan Muhammad , Alshareef Sohad , Sheikh Huda , Alaryani Fatima , AL-Kattan Manal , Alatawi Hanan , Menaa Farid , Nadeem Aroosa
Published: October 2022

Resin composites have been widely used in dental restoration. However, polymerization shrinkage and resultant bacterial microleakage are major limitations that may lead to secondary caries. To overcome this, a new type of antibacterial resin composite containing ciprofloxacin-loaded silver nanoparticles (CIP-AgNPs) were synthesized. The chemical reduction approach successfully produced CIP-AgNPs, as demonstrated by FTIR, zeta potential, scanning electron microscopy, and ultraviolet-visible (UV-vis) spectroscopy. CIP-AgNPs were added to resin composites and the antibacterial activity of the dental composite discs were realized against Enterococcus faecalis, Streptococcus mutans, and the Saliva microcosm. The biocompatibility of modified resin composites was assessed and mechanical testing of modified dental composites was also performed. The results indicated that the antibacterial activity and compressive strength of resin composites containing CIP-AgNPs were enhanced compared to the control group. They were also biocompatible when compared to resin composites containing AgNPs. In short, these results established strong ground application for CIP-AgNP-modified dental composite resins.
Analysis of Changes in the Amount of Phytosterols after the Bleaching Process of Hemp Oils
Authors: Kwaśnica Andrzej , Teleszko Mirosława , Marcinkowski Damian , Kmiecik Dominik , Grygier Anna , Golimowski Wojciech
Published: October 2022

Unrefined vegetable oils from niche oilseeds are now sought after by consumers because of their unique nutritional properties and taste qualities. The color and flavor intensity of niche oils is a big problem, and their refining is not industrially feasible due to the small production scale. The study undertaken aimed analyze the effect of changing the amount of phytosterols (PSs) after the bleaching process of hemp oils of the ‘Finola’, ‘Earlina 8FC’ and ‘Secuieni Jubileu’ varieties. Cold-pressed (C) and hot-pressed (H) crude vegetable oils were bleached with selected bleaching earth (BE) at two concentrations. The post-process BE was extracted with methanol. The amount of PSs in the crude oils and the extract after washing the BE with methanol was analyzed by GC (gas chromatography). The study shows that the bleaching process did not significantly affect the depletion of PSs in the oils. Trace amounts of PSs remain on the BE and, due to methanol extraction, can be extracted from the oil. The conclusion of the performed research is that the bleaching of hemp oil does not cause depletion of the oil, and it significantly improves organoleptic properties. The oil bleaching process results in an oil loss of less than 2% by weight of the bleached oil, while the loss depends on the type of BE used. The study shows that the loss of phytosterols after the bleaching process averages 2.69 ± 0.69%, and depends on the type of BE used and the oil extracted from different varieties of hemp seeds.