bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effects of Intravitreal Administration of Antifungal Drugs on the Structure and Mechanical Properties Peripheral Blood Erythrocyte Surface in Rabbits
Authors: Starodubtseva Maria N. , Karachrysafi Sofia , Shkliarava Nastassia M. , Chelnokova Irina A. , Kavvadas Dimitrios , Papadopoulou Kyriaki , Samara Paraskevi , Papaliagkas Vasileios , Sioga Antonia , Komnenou Anastasia , Karampatakis Vasileios , Papamitsou Theodora
Published: September 2022

Background: Fungal infections can pose great threat to sight. Immediate treatment is usually required; antifungal agents are widely accepted and are effective in most cases. The present experimental study aims to investigate the probable effects of intravitreal injection of antifungal agents on the structure and mechanical properties of the surface of peripheral blood erythrocytes. Methods: Nine albino New Zealand white rabbits, aged five months old, were chosen for the experiment. Solutions of micafungin, voriconazole, or balanced salt solution (BSS) were injected into the midvitreous. Animals were divided into two experimental groups and one control group. Blood sampling from an intravenous (IV) line was performed after 10 days from the last IV injection. An atomic force microscope (AFM) was used to study the structural and mechanical properties of cell surfaces. Results: The analysis results showed that the parameters of the cytoskeleton’s spatial organization changed insignificantly with the antifungal drug treatment. Conclusions: Our findings suggest that locally administered antifungal drugs can cause significant changes to the structure and frictional properties of the erythrocyte surface. These effects occur in the long-term period after administration of the drugs and represent a potential possibility for violation of blood supply to tissues, and the further development of negative side effects.
Atomic Simulation of the Binding of JAK1 and JAK2 with the Selective Inhibitor Ruxolitinib
Authors: Kondratyev Maxim , Rudnev Vladimir R. , Nikolsky Kirill S. , Stepanov Alexander A. , Petrovsky Denis V. , Kulikova Liudmila I. , Kopylov Arthur T. , Malsagova Kristina A. , Kaysheva Anna L.
Published: September 2022

Rheumatoid arthritis belongs to the group of chronic systemic autoimmune diseases characterized by the development of destructive synovitis and extra-articular manifestations. Cytokines regulate a wide range of inflammatory processes involved in the pathogenesis of rheumatoid arthritis and contribute to the induction of autoimmunity and chronic inflammation. Janus-associated kinase (JAK) and signal transducer and activator of transcription (STAT) proteins mediate cell signaling from cytokine receptors, and are involved in the pathogenesis of autoimmune and inflammatory diseases. Targeted small-molecule drugs that inhibit the functional activity of JAK proteins are used in clinical practice for the treatment of rheumatoid arthritis. In our study, we modeled the interactions of the small-molecule drug ruxolitinib with JAK1 and JAK2 isoforms and determined the binding selectivity using molecular docking. Molecular modeling data show that ruxolitinib selectively binds the JAK1 and JAK2 isoforms with a binding affinity of −8.3 and −8.0 kcal/mol, respectively. The stabilization of ligands in the cavity of kinases occurs primarily through hydrophobic interactions. The amino acid residues of the protein globules of kinases that are responsible for the correct positioning of the drug ruxolitinib and its retention have been determined.
Senescence-Independent Anti-Inflammatory Activity of the Senolytic Drugs Dasatinib, Navitoclax, and Venetoclax in Zebrafish Models of Chronic Inflammation
Authors: Hernández-Silva David , Cantón-Sandoval Joaquín , Martínez-Navarro Francisco Juan , Pérez-Sánchez Horacio , de Oliveira Sofia , Mulero Victoriano , Alcaraz-Pérez Francisca , Cayuela María Luisa
Published: September 2022

Telomere shortening is the main molecular mechanism of aging, but not the only one. The adaptive immune system also ages, and older organisms tend to develop a chronic pro-inflammatory status with low-grade inflammation characterized by chronic activation of the innate immune system, called inflammaging. One of the main stimuli that fuels inflammaging is a high nutrient intake, triggering a metabolic inflammation process called metainflammation. In this study, we report the anti-inflammatory activity of several senolytic drugs in the context of chronic inflammation, by using two different zebrafish models: (i) a chronic skin inflammation model with a hypomorphic mutation in spint1a, the gene encoding the serine protease inhibitor, kunitz-type, 1a (also known as hai1a) and (ii) a non-alcoholic fatty liver disease/non-alcoholic steatohepatitis (NAFLD/NASH) model with inflammation induced by a high-fat diet. Our results show that, although these models do not manifest premature aging, the senolytic drugs dasatinib, navitoclax, and venetoclax have an anti-inflammatory effect that results in the amelioration of chronic inflammation.
Potent Chlorambucil-Platinum(IV) Prodrugs
Authors: Aputen Angelico D. , Elias Maria George , Gilbert Jayne , Sakoff Jennette A. , Gordon Christopher P. , Scott Kieran F. , Aldrich-Wright Janice R.
Published: September 2022

The DNA-alkylating derivative chlorambucil was coordinated in the axial position to atypical cytotoxic, heterocyclic, and non-DNA coordinating platinum(IV) complexes of type, [PtIV(HL)(AL)(OH)2](NO3)2 (where HL is 1,10-phenanthroline, 5-methyl-1,10-phenanthroline or 5,6-dimethyl-1,10-phenanthroline, AL is 1S,2S-diaminocyclohexane). The resultant platinum(IV)-chlorambucil prodrugs, PCLB, 5CLB, and 56CLB, were characterized using high-performance liquid chromatography, nuclear magnetic resonance, ultraviolet-visible, circular dichroism spectroscopy, and electrospray ionization mass spectrometry. The prodrugs displayed remarkable antitumor potential across multiple human cancer cell lines compared to chlorambucil, cisplatin, oxaliplatin, and carboplatin, as well as their platinum(II) precursors, PHENSS, 5MESS, and 56MESS. Notably, 56CLB was exceptionally potent in HT29 colon, Du145 prostate, MCF10A breast, MIA pancreas, H460 lung, A2780, and ADDP ovarian cell lines, with GI50 values ranging between 2.7 and 21 nM. Moreover, significant production of reactive oxygen species was detected in HT29 cells after treatment with PCLB, 5CLB, and 56CLB up to 72 h compared to chlorambucil and the platinum(II) and (IV) precursors.
Dietary Modulation of the Human Gut Microbiota and Metabolome with Flaxseed Preparations
Authors: Kleigrewe Karin , Haack Martina , Baudin Martine , Ménabréaz Thomas , Crovadore Julien , Masri Mahmoud , Beyrer Michael , Andlauer Wilfried , Lefort François , Dawid Corinna , Brück Thomas B. , Brück Wolfram M.
Published: September 2022

Flaxseeds are typically consumed either as whole flaxseed, ground flaxseed, flaxseed oil, partially defatted flaxseed meal, or as a milk alternative. They are considered a rich source of vitamins, minerals, proteins and peptides, lipids, carbohydrates, lignans, and dietary fiber, which have shown hypolipidemic, antiatherogenic, anticholesterolemic, and anti-inflammatory property activity. Here, an in vitro batch culture model was used to investigate the influence of whole milled flaxseed and partially defatted milled flaxseed press cake on the gut microbiota and the liberation of flaxseed bioactives. Microbial communities were profiled using 16S rRNA gene-based high-throughput sequencing with targeted mass spectrometry measuring lignan, cyclolinopeptide, and bile acid content and HPLC for short-chain fatty acid profiles. Flaxseed supplementation decreased gut microbiota richness with Firmicutes, Proteobacteria, and Bacteroidetes becoming the predominant phyla. Secoisolariciresinol, enterodiol, and enterolactone were rapidly produced with acetic acid, butyric acid, and propionic acid being the predominant acids after 24 h of fermentation. The flaxseed press cake and whole flaxseed were equivalent in microbiota changes and functionality. However, press cake may be superior as a functional additive in a variety of foods in terms of consumer acceptance as it would be more resistant to oxidative changes.
Cloning and Characterization of Fructose-1,6-Bisphosphate Aldolase from Euphausia superba
Authors: Xia Jikun , Xin Wanmeng , Wang Fang , Xie Wancui , Liu Yi , Xu Jiakun
Published: September 2022

Fructose-1,6-bisphosphate aldolase (EC 4.1.2.13) is a highly conserved enzyme that is involved in glycolysis and gluconeogenesis. In this study, we cloned the fructose-1,6-bisphosphate aldolase gene from Euphausia superba (EsFBA). The full-length cDNA sequence of EsFBA is 1098 bp long and encodes a 365-amino-acid protein. The fructose-1,6-bisphosphate aldolase gene was expressed in Escherichia coli (E. coli). A highly purified protein was obtained using HisTrap HP affinity chromatography and size-exclusion chromatography. The predicted three-dimensional structure of EsFBA showed a 65.66% homology with human aldolase, whereas it had the highest homology (84.38%) with the FBA of Penaeus vannamei. Recombinant EsFBA had the highest activity at 45 °C and pH 7.0 in phosphate buffer. By examining the activity of metal ions and EDTA, we found that the effect of metal ions and EDTA on EsFBA’s enzyme activity was not significant, while the presence of borohydride severely reduced the enzymatic activity; thus, EsFBA was confirmed to be a class I aldolase. Furthermore, targeted mutations at positions 34, 147, 188, and 230 confirmed that they are key amino acid residues for EsFBA.
Integrating Genetic and Chromosome Maps of Allium cepa: From Markers Visualization to Genome Assembly Verification
Authors: Ermolaev Aleksey , Kudryavtseva Natalia , Pivovarov Anton , Kirov Ilya , Karlov Gennady , Khrustaleva Ludmila
Published: September 2022

The ability to directly look into genome sequences has opened great opportunities in plant breeding. Yet, the assembly of full-length chromosomes remains one of the most difficult problems in modern genomics. Genetic maps are commonly used in de novo genome assembly and are constructed on the basis of a statistical analysis of the number of recombinations. This may affect the accuracy of the ordering and orientation of scaffolds within the chromosome, especially in the region of recombination suppression. Moreover, it is impossible to assign contigs lacking DNA markers. Here, we report the use of Tyr-FISH to determine the position of the short DNA sequence of markers and non-mapped unique copy sequence on the physical chromosomes of a large-genome onion (Allium cepa L.). In order to minimize potential background masking of the target signal, we improved our earlier developed pipeline for probe design. A total of 23 markers were located on physical chromosomes 2 and 6. The order of markers was corrected by the integration of genetic, pseudochromosome maps and cytogenetic maps. Additionally, the position of the mlh1 gene, which was not on the genetic map, was defined on physical chromosome 2. Tyr-FISH mapping showed that the order of 23.1% (chromosome 2) and 27.3% (chromosome 6) of the tested genes differed between physical chromosomes and pseudochromosomes. The results can be used for the improvement of pseudochromosome 2 and 6 assembly. The present study aims to demonstrate the value of the in situ visualization of DNA sequences in chromosome-scaffold genome assembly.
New Insights on Heme Uptake in Leishmania spp.
Authors: Cabello-Donayre María , Orrego Lina M. , Herráez Elisa , García-Hernández Raquel , Pérez-Victoria José M.
Published: September 2022

The protozoan parasite Leishmania, responsible for leishmaniasis, is one of the few aerobic organisms that cannot synthesize the essential molecule heme. Therefore, it has developed specialized pathways to scavenge it from its host. In recent years, some proteins involved in the import of heme, such as LHR1 and LFLVCRB, have been identified, but relevant aspects regarding the process remain unknown. Here, we characterized the kinetics of the uptake of the heme analogue Zn(II) Mesoporphyrin IX (ZnMP) in Leishmania major promastigotes as a model of a parasite causing cutaneous leishmaniasis with special focus on the force that drives the process. We found that ZnMP uptake is an active, inducible, and pH-dependent process that does not require a plasma membrane proton gradient but requires the presence of the monovalent cations Na+ and/or K+. In addition, we demonstrated that this parasite can efflux this porphyrin against a concentration gradient. We also found that ZnMP uptake differs among different dermotropic or viscerotropic Leishmania species and does not correlate with LHR1 or LFLVCRB expression levels. Finally, we showed that these transporters have only partially overlapping functions. Altogether, these findings contribute to a deeper understanding of an important process in the biology of this parasite.
Current Uses of Mushrooms in Cancer Treatment and Their Anticancer Mechanisms
Authors: Park Hye-Jin
Published: September 2022

Cancer is the leading cause of mortality worldwide. Various chemotherapeutic drugs have been extensively used for cancer treatment. However, current anticancer drugs cause severe side effects and induce resistance. Therefore, the development of novel and effective anticancer agents with minimal or no side effects is important. Notably, natural compounds have been highlighted as anticancer drugs. Among them, many researchers have focused on mushrooms that have biological activities, including antitumor activity. The aim of this review is to discuss the anticancer potential of different mushrooms and the underlying molecular mechanisms. We provide information regarding the current clinical status and possible modes of molecular actions of various mushrooms and mushroom-derived compounds. This review will help researchers and clinicians in designing evidence-based preclinical and clinical studies to test the anticancer potential of mushrooms and their active compounds in different types of cancers.
Scattered Tubular Cells Markers in Macula Densa of Normal Human Adult Kidney
Authors: Tossetta Giovanni , Fantone Sonia , Lorenzi Teresa , Galosi Andrea Benedetto , Sagrati Andrea , Fabri Mara , Marzioni Daniela , Morroni Manrico
Published: September 2022

Background: The scattered tubular cells (STCs) are a population of resident progenitor tubular cells with expansion, self-renewal and epithelial differentiation abilities. Although these cells are localized within the proximal (PTs) and distal (DTs) tubules in a normal adult kidney, their presence has never been demonstrated in human macula densa (MD). The purpose of the present study is to describe the presence of STCs in MD using specific markers such as prominin-1 (CD133), cytokeratin 7 (KRT7) and vimentin (VIM). Methods: We analyzed two sets of three consecutive serial sections for each sample. The first sections of each set were immunostained for nNOS to identify MD, the second sections were immune-stained for CD133 (specific STCs marker) while the third sections were analyzed for KRT7 (another STCs specific marker) and VIM (that stains the basal pole of the STCs) in the first and second sets, respectively, in order to study the co-expression of KRT7 and VIM with the CD133 marker. Results: CD133 was localized in some MD cells and in the adjacent DT cells. Moreover, CD133 was detected in the parietal epithelial cells of Bowman’s capsule and in some proximal tubules (PT). KRT7-positive cells were identified in MD and adjacent DT cells, while KRT7 positivity was mostly confined in both DT and collecting ducts (CD) in the other areas of the renal parenchyma. CD133 and KRT7 were co-expressed in some MD and adjacent DT cells. Some of the latter cells were positive both for CD133 and VIM. CD133 was always localized in the apical part of the cells, whereas the VIM expression was evident only in the cellular basal pole. Although some cells of MD expressed VIM or CD133, none of them co-expressed VIM and CD133. Conclusions: The presence of STCs was demonstrated in human adult MD, suggesting that this structure has expansion, self-renewal and epithelial differentiation abilities, similar to all other parts of renal tubules.
Development of Soft Rice Lines by Regulating Amylose Content via Editing the 5′UTR of the Wx Gene
Authors: Yang Jinlian , Guo Xinying , Wang Xuan , Fang Yaoyu , Liu Fang , Qin Baoxiang , Li Rongbai
Published: September 2022

The type of soft rice with low amylose content (AC) is more and more favored by consumers for its better eating and cooking quality, as people’s quality of life continuously improves in China. The Wx gene regulates the AC of rice grains, thus affecting the degree of softness of the rice. Mei Meng B (MMB), Tian Kang B (TKB), and DR462 are three indica rice maintained lines with good morphological characters, but also with undesirably high AC. Therefore, CRISPR/Cas9 technology was used to edit the Wx gene of these lines to create a batch of soft rice breeding materials. New gene-edited lines MMB-10-2, TKB-21-12, and DR462-9-9, derived from the above parental lines, respectively, were selected in the T2 generations, with an AC of 17.2%, 16.8%, and 17.8%, and gel consistency (GC) of 78.6 mm, 77.4 mm, and 79.6 mm, respectively. The rapid viscosity analysis (RVA) spectrum showed that the three edited lines had a better eating quality as compared to the corresponding wild type, and showing new characteristics, different from the high-quality soft rice popular in the market. There was no significant difference in the main agronomic traits in the three edited lines compared to the corresponding wild types. Moreover, the chalkiness of DR462-9-9 was reduced, resulting in an improved appearance of its polished rice. The present study created soft rice germplasms for breeding improved quality hybrid rice, without changing the excellent traits of their corresponding wild type varieties.
Methods of the Large-Scale Production of Extracellular Vesicles
Authors: Syromiatnikova Valeriia , Prokopeva Angelina , Gomzikova Marina
Published: September 2022

To date, extracellular vesicles (EVs) have been extensively investigated as potential substitutes for cell therapy. Research has suggested their ability to overcome serious risks associated with the application of these cells. Although, the translation of EVs into clinical practice is hampered by the lack of a cheap reasonable way to obtain a clinically relevant number of EVs, an available method for the large-scale production of EVs ensures vesicles’ integrity, preserves their biological activity, and ensures they are well reproducible, providing homogeneity of the product from batch to batch. In this review, advances in the development of methods to increase EVs production are discussed. The existing approaches can be divided into the following: (1) those based on increasing the production of natural EVs by creating and using high capacity “cell factories”, (2) those based on the induction of EVs secretion under various cell stressors, and (3) those based on cell fragmentation with the creation of biomimetic vesicles. The aim of this review is to stimulate the introduction of EVs into clinical practice and to draw attention to the development of new methods of EVs production on a large scale.
The Role of microRNAs in the Gonocyte Theory as Target of Malignancy: Looking for Potential Diagnostic Biomarkers
Authors: García-Andrade Fabiola , Vigueras-Villaseñor Rosa María , Chávez-Saldaña Margarita Dolores , Rojas-Castañeda Julio César , Bahena-Ocampo Iván Uriel , Aréchaga-Ocampo Elena , Díaz-Chávez José , Landero-Huerta Daniel Adrian
Published: September 2022

Some pediatric patients with cryptorchidism preserve cells with gonocyte characteristics beyond their differentiation period, which could support the theory of the gonocyte as a target for malignancy in the development of testicular neoplasia. One of the key molecules in gonocyte malignancy is represented by microRNAs (miRNAs). The goal of this review is to give an overview of miRNAs, a class of small non-coding RNAs that participate in the regulation of gene expression. We also aim to review the crucial role of several miRNAs that have been further described in the regulation of gonocyte differentiation to spermatogonia, which, when transformed, could give rise to germ cell neoplasia in situ, a precursor lesion to testicular germ cell tumors. Finally, the potential use of miRNAs as diagnostic and prognostic biomarkers in testicular neoplasia is addressed, due to their specificity and sensitivity compared to conventional markers, as well as their applications in therapeutics.
Ustilaginoidea virens Nuclear Effector SCRE4 Suppresses Rice Immunity via Inhibiting Expression of a Positive Immune Regulator OsARF17
Authors: Qiu Shanshan , Fang Anfei , Zheng Xinhang , Wang Shanzhi , Wang Jiyang , Fan Jing , Sun Zongtao , Gao Han , Yang Jiyun , Zeng Qingtao , Cui Fuhao , Wang Wen-Ming , Chen Jianping , Sun Wenxian
Published: September 2022

Rice false smut caused by the biotrophic fungal pathogen Ustilaginoidea virens has become one of the most important diseases in rice. The large effector repertory in U. virens plays a crucial role in virulence. However, current knowledge of molecular mechanisms how U. virens effectors target rice immune signaling to promote infection is very limited. In this study, we identified and characterized an essential virulence effector, SCRE4 (Secreted Cysteine-Rich Effector 4), in U. virens. SCRE4 was confirmed as a secreted nuclear effector through yeast secretion, translocation assays and protein subcellular localization, as well as up-regulation during infection. The SCRE4 gene deletion attenuated the virulence of U. virens to rice. Consistently, ectopic expression of SCRE4 in rice inhibited chitin-triggered immunity and enhanced susceptibility to false smut, substantiating that SCRE4 is an essential virulence factor. Furthermore, SCRE4 transcriptionally suppressed the expression of OsARF17, an auxin response factor in rice, which positively regulates rice immune responses and resistance against U. virens. Additionally, the immunosuppressive capacity of SCRE4 depended on its nuclear localization. Therefore, we uncovered a virulence strategy in U. virens that transcriptionally suppresses the expression of the immune positive modulator OsARF17 through nucleus-localized effector SCRE4 to facilitate infection.
Identification of Candidate Chemosensory Gene Families by Head Transcriptomes Analysis in the Mexican Fruit Fly, Anastrepha ludens Loew (Diptera: Tephritidae)
Authors: Segura-León Obdulia L. , Torres-Huerta Brenda , Estrada-Pérez Alan Rubén , Cibrián-Tovar Juan , Hernandez-Hernandez Fidel de la Cruz , Cruz-Jaramillo José Luis , Meza-Hernández José Salvador , Sánchez-Galicia Fabian
Published: September 2022

Insect chemosensory systems, such as smell and taste, are mediated by chemosensory receptor and non-receptor protein families. In the last decade, many studies have focused on discovering these families in Tephritidae species of agricultural importance. However, to date, there is no information on the Mexican fruit fly Anastrepha ludens Loew, a priority pest of quarantine importance in Mexico and other countries. This work represents the first effort to identify, classify and characterize the six chemosensory gene families by analyzing two head transcriptomes of sexually immature and mature adults of A. ludens from laboratory-reared and wild populations, respectively. We identified 120 chemosensory genes encoding 31 Odorant-Binding Proteins (OBPs), 5 Chemosensory Proteins (CSPs), 2 Sensory Neuron Membrane Proteins (SNMPs), 42 Odorant Receptors (ORs), 17 Ionotropic Receptors (IRs), and 23 Gustatory Receptors (GRs). The 120 described chemosensory proteins of the Mexican fruit fly significantly contribute to the genetic databases of insects, particularly dipterans. Except for some OBPs, this work reports for the first time the repertoire of olfactory proteins for one species of the genus Anastrepha, which provides a further basis for studying the olfactory system in the family Tephritidae, one of the most important for its economic and social impact worldwide.
Assembly of the Multi-Subunit Cytochrome bc1 Complex in the Yeast Saccharomyces cerevisiae
Authors: Zara Vincenzo , De Blasi Gabriella , Ferramosca Alessandra
Published: September 2022

The cytochrome bc1 complex is an essential component of the mitochondrial respiratory chain of the yeast Saccharomyces cerevisiae. It is composed of ten protein subunits, three of them playing an important role in electron transfer and proton pumping across the inner mitochondrial membrane. Cytochrome b, the central component of this respiratory complex, is encoded by the mitochondrial genome, whereas all the other subunits are of nuclear origin. The assembly of all these subunits into the mature and functional cytochrome bc1 complex is therefore a complicated process which requires the participation of several chaperone proteins. It has been found that the assembly process of the mitochondrial bc1 complex proceeds through the formation of distinct sub-complexes in an ordered sequence. Most of these sub-complexes have been thoroughly characterized, and their molecular compositions have also been defined. This study critically analyses the results obtained so far and highlights new possible areas of investigation.
Action of the Photochrome Glyght on GABAergic Synaptic Transmission in Mouse Brain Slices
Authors: Petukhova Elena , Ponomareva Daria , Rustler Karin , Koenig Burkhard , Bregestovski Piotr
Published: September 2022

Glyght is a new photochromic compound described as an effective modulator of glycine receptors at heterologous expression, in brain slices and in zebrafish larvae. Glyght also caused weak inhibition of GABAA-mediated currents in a cell line expressing α1/β2/γ2 GABAA receptors. However, the effects of Glyght on GABAergic transmission in the brain have not been analysed, which does not allow a sufficiently comprehensive assessment of the effects of the compound on the nervous system. Therefore, in this study using whole-cell patch-clamp recording, we analysed the Glyght (100 µM) action on evoked GABAergic inhibitory postsynaptic currents (eIPSCs) in mice hippocampal slices. Two populations of cells were found: the first responded by reducing the GABAergic eIPSCs’ amplitude, whereas the second showed no sensitivity to the compound. Glyght did not affect the ionic currents’ amplitude induced by GABA application, suggesting the absence of action on postsynaptic GABA receptors. Additionally, Glyght had no impact on the paired-pulse modulation of GABAergic eIPSCs, indicating that Glyght does not modulate the neurotransmitter release mechanisms. In the presence of strychnine, an antagonist of glycine receptors, the Glyght effect on GABAergic synaptic transmission was absent. Our results suggest that Glyght can modulate GABAergic synaptic transmission via action on extrasynaptic glycine receptors.
Targeting PSEN1 by lnc-CYP3A43-2/miR-29b-2-5p to Reduce β Amyloid Plaque Formation and Improve Cognition Function
Authors: Wuli Wei , Lin Shinn-Zong , Chen Shee-Ping , Tannous Bakhos A. , Huang Wen-Sheng , Woon Peng Yeong , Wu Yang-Chang , Yang Hsueh-Hui , Chen Yi-Cheng , Fleming Renata Lopes , Rogers Jack T. , Cahill Catherine M. , Ho Tsung-Jung , Chiou Tzyy-Wen , Harn Horng-Jyh
Published: September 2022

Presenilin-1 (PSEN1) is a crucial subunit within the γ-secretase complex and regulates β-amyloid (Aβ) production. Accumulated evidence indicates that n-butylidenephthalide (BP) acts effectively to reduce Aβ levels in neuronal cells that are derived from trisomy 21 (Ts21) induced pluripotent stem cells (iPSCs). However, the mechanism underlying this effect remains unclear. This article aims to investigate the possible mechanisms through which BP ameliorates the development of Alzheimer’s disease (AD) and verify the effectiveness of BP through animal experiments. Results from RNA microarray analysis showed that BP treatment in Ts21 iPSC-derived neuronal cells reduced long noncoding RNA (lncRNA) CYP3A43-2 levels and increased microRNA (miR)-29b-2-5p levels. Bioinformatics tool prediction analysis, biotin-labeled miR-29b-2-5p pull-down assay, and dual-luciferase reporter assay confirmed a direct negative regulatory effect for miRNA29b-2-5p on lnc-RNA-CYP3A43-2 and PSEN1. Moreover, BP administration improved short-term memory and significantly reduced Aβ accumulation in the hippocampus and cortex of 3xTg-AD mice but failed in miR-29b-2-5p mutant mice generated by CRISP/Cas9 technology. In addition, analysis of brain samples from patients with AD showed a decrease in microRNA-29b-2-5p expression in the frontal cortex region. Our results provide evidence that the LncCYP3A43-2/miR29-2-5p/PSEN1 network might be involved in the molecular mechanisms underlying BP-induced Aβ reduction.
SPIN90 Deficiency Ameliorates Amyloid β Accumulation by Regulating APP Trafficking in AD Model Mice
Authors: Oh Youngsoo , Lee Wongyoung , Kim So Hee , Lee Sooji , Kim Byeong C. , Lee Kun Ho , Kim Sung Hyun , Song Woo Keun
Published: September 2022

Alzheimer’s disease (AD), a common form of dementia, is caused in part by the aggregation and accumulation in the brain of amyloid β (Aβ), a product of the proteolytic cleavage of amyloid precursor protein (APP) in endosomes. Trafficking of APP, such as surface-intracellular recycling, is an early critical step required for Aβ generation. Less is known, however, about the molecular mechanism regulating APP trafficking. This study investigated the mechanism by which SPIN90, along with Rab11, modulates APP trafficking, Aβ motility and accumulation, and synaptic functionality. Brain Aβ deposition was lower in the progeny of 5xFAD-SPIN90KO mice than in 5xFAD-SPIN90WT mice. Analysis of APP distribution and trafficking showed that the surface fraction of APP was locally distinct in axons and dendrites, with these distributions differing significantly in 5xFAD-SPIN90WT and 5xFAD-SPIN90KO mice, and that neural activity-driven APP trafficking to the surface and intracellular recycling were more actively mobilized in 5xFAD-SPIN90KO neurons. In addition, SPIN90 was found to be cotrafficked with APP via axons, with ablation of SPIN90 reducing the intracellular accumulation of APP in axons. Finally, synaptic transmission was restored over time in 5xFAD-SPIN90KO but not in 5xFAD-SPIN90WT neurons, suggesting SPIN90 is implicated in Aβ production through the regulation of APP trafficking.
Affinities to Oxaliplatin: Vitamins from B Group vs. Nucleobases
Authors: Szefler Beata , Czeleń Przemysław , Wojtkowiak Kamil , Jezierska Aneta
Published: September 2022

Oxaliplatin, similar to Cisplatin, exhibits anticancer activity by interacting with DNA and inducing programmed cell death. It is biotransformed through a number of spontaneous and non-enzymatic processes. In this way, several transient reactive species are formed, including dichloro-, monochloro-, and diaqua-DACH platin, which can complex with DNA and other macromolecules. The molecular level suggests that such interactions can also take place with vitamins containing aromatic rings with lone pair orbitals. Theoretical and experimental studies were performed to investigate interactions of vitamins from the B group with Oxaliplatin, and the results were compared with values characterizing native purines. Quantum-chemical simulations were carried out at the B3LYP/6-31G(d,p) level, with the LANL2DZ basis set representing atomic orbitals of platinum atom, and at the MN15/def2-TZVP levels of theory with the use of Polarizable Continuum Model (IEF-PCM formulation) and water as a solvent. Additionally, time-dependent density functional theory (TD-DFT) was employed to study molecular properties in the electronic excited state. Interactions of vitamins and Oxaliplatin were investigated using UV-Vis spectroscopy. Values of the free energy (ΔGr) indicate spontaneous reactions with monoaqua [PtH2OClDACH]+ and diaqua [Pt(H2O)2DACH]2+ derivatives of Oxaliplatin. However, diaqua derivatives were found to be preferable. The free energy (ΔGr) values obtained for vitamins from the B group indicate lower affinity of Oxaliplatin compared with values characterizing complexes formed by guanine, adenine, and cytosine. The exception is the monoaqua form of vitamin B1 (thiamine) at the MN15/def2-TZVP levels of calculations. An application of atoms in molecules (AIM) theory revealed non-covalent interactions present in the complexes studied. The comparison of computed and experimental spectroscopic properties showed a good agreement.
Genome-Wide Identification of WRKY Gene Family in Pitaya Reveals the Involvement of HmoWRKY42 in Betalain Biosynthesis
Authors: Chen Canbin , Xie Fangfang , Shah Kamran , Hua Qingzhu , Chen Jiayi , Zhang Zhike , Zhao Jietang , Hu Guibing , Qin Yonghua
Published: September 2022

The WRKY gene family is a plant-specific transcription factor (TF) that regulates many physiological processes and (a) biotic stress responses. Despite this, little is known about the molecular properties and roles of WRKY TFs in pitaya betalain biosynthesis. Here we report the identification of 70 WRKY in Hylocereus undatus, their gene structure, locations on each chromosome, systematic phylogenetic analysis, conserved motif analysis, and synteny of HuWRKY genes. HmoWRKY42 is a Group IIb WRKY protein and contains a coiled-coil motif, a WRKY domain and a C2H2 zinc-finger motif (CX5CX23HXH). Results from yeast one-hybrid and transient dual-luciferase assays showed that HmoWRKY42 was a transcriptional repressor and could repress HmocDOPA5GT1 expression by binding to its promoter. Yeast two-hybrid assays showed that HmoWRKY42 could interact with itself to form homodimers. Knocking out the coiled-coil motif of HmoWRKY42 prevented its self-interaction and prevented it from binding to the HmocDOPA5GT1 promoter. Knocking out the WRKY domain and C2H2 zinc-finger motif sequence of HmoWRKY42 also prevented it from binding to the HmocDOPA5GT1 promoter. The coiled-coil motif, the WRKY domain and the C2H2 zinc finger motif are key motifs for the binding of HmoWRKY42 to the HmocDOPA5GT1 promoter. HmoWRKY42 is localized in the nucleus and possesses trans-activation ability responsible for pitaya betalain biosynthesis by repressing the transcription of HmocDOPA5GT1. As far as we know, no reports are available on the role of HmoWRKY42 in pitaya betalain biosynthesis. The results provide an important foundation for future analyses of the regulation and functions of the HuWRKY gene family.
GoTO: A Process-Navigation Tool for Telehealth and -Care Solutions, Designed to Ensure an Efficient Trajectory from Goal Setting to Outcome Evaluation
Authors: Kayser Lars , Furstrand Dorthe , Nyman Rasmussen Emil , Monberg Ann-Catrine , Karnoe Astrid
Published: September 2022

Objectives: The digital transformation of the healthcare sector involves the procurement and implementation of new health technologies, which will likely be a challenge to healthcare providers who are not part of large organizations. In response to the needs of small and middle-sized health and care organizations, we have developed a process navigator to guide providers of healthcare through the processes of innovation, the procurement of mature products, and their implementation in telehealth and telecare projects. Methods: A narrative overview identified health-technology-assessment-inspired models. Conversations with national and international colleagues identified project and implementation models. The origin of the included models was identified, and relevant articles were referred to to describe the essential principles, including the nature of stakeholder involvement and the evaluation processes when appropriate. Based on the inputs, we proposed the process navigator GoTO. Results: Six health-technology-assessment-inspired models, six project models, one implementation model, and one innovation model were identified and informed the creation of the GoTO process navigator. The navigator consists of four parts: inception (eight steps); materialization (three tracks, depending on the maturity of the planned solution); implementation (five steps); and the final assessment and evaluation. Conclusion: The GoTO process navigator is an intuitive guide for innovation, procurement, and implementation in telehealth and -care. The GoTo navigator can assist providers of digital health and care services throughout the process from the initial identification of goals to the final evaluation of outcomes.
Annual Fluctuations in Winter Colony Losses of Apis mellifera L. Are Predicted by Honey Flow Dynamics of the Preceding Year
Authors: Johannesen Jes , Wöhl Saskia , Berg Stefen , Otten Christoph
Published: September 2022

Winter loss rates of honey bee colonies may fluctuate highly between years in temperate climates. The present study combined survey data of autumn and winter loss rates in Germany (2012–2021) with estimates of honey flow—assessed with automated hive scales as the start of honey flow in spring and its magnitude in summer—with the aim of understanding annual fluctuations in loss rates. Autumn colony loss rates were positively and significantly correlated with winter loss rates, whereas winter loss rates were inversely related to loss rates in autumn of the following year. An early start of net honey flow in spring predicted high loss rates in both autumn and winter, whereas high cumulative honey flow led to lower loss rates. The start of net honey flow was related to temperature sums in March. Combined, the results implied that the winter loss rate in one year was influenced by the loss rate of the preceding winter and shaped by honey flow dynamics during the following year. Hence, the rate of colony loss in winter can be viewed as a cumulative death process affected by the preceding one and a half years.
Evaluation of Left Ventricular Function Using Four-Dimensional Flow Cardiovascular Magnetic Resonance: A Systematic Review
Authors: Qin Jiaxing Jason , Indja Ben , Gholipour Alireza , Gök Mustafa , Grieve Stuart M.
Published: September 2022

There is increasing recognition of the value of four-dimensional flow cardiovascular magnetic resonance (4D-flow MRI) as a potential means to detect and measure abnormal flow behaviour that occurs during early left ventricular (LV) dysfunction. We performed a systematic review of current literature on the role of 4D-flow MRI-derived flow parameters in quantification of LV function with a focus on potential clinical applicability. A comprehensive literature search was performed in March 2022 on available databases. A total of 1186 articles were identified, and 30 articles were included in the final analysis. All the included studies were ranked as “highly clinically applicable”. There was considerable variability in the reporting of methodologies and analyses. All the studies were small-scale feasibility or pilot studies investigating a diverse range of flow parameters. The most common primary topics of investigation were energy-related flow parameters, flow components and vortex analysis which demonstrated potentials for quantifying early diastolic dysfunction, whilst other parameters including haemodynamic forces, residence time distribution and turbulent kinetic energy remain in need of further evaluation. Systematic quantitative comparison of study findings was not possible due to this heterogeneity, therefore limiting the collective power of the studies in evaluating clinical applicability of the flow parameters. To achieve broader clinical application of 4D-flow MRI, larger scale investigations are required, together with standardisation of methodologies and analytical approach.
Traumatic Brain Injury Induces a Differential Immune Response in Polytrauma Patients; Prospective Analysis of CD69 Expression on T Cells and Platelet Expansion
Authors: Ditsch Alexander , Hunold Lea , Hefele Friederike , Greve Frederik , Mair Olivia , Biberthaler Peter , Heimann Laura , Hanschen Marc
Published: September 2022

Background: Accidents and injuries are the leading causes of mortality in young people. CD4+ regulatory T cells (CD4+ Tregs), Th17 cells and platelets could be identified as key players in post-traumatic immunological dysfunction, which is a common cause of late mortality in trauma patients. The mechanisms of activation of these cell types and their interaction remain mostly unclear. Since CD69 is not only a leukocyte marker but has also immunoregulatory functions, we postulate a role for CD69 after trauma. The present study investigates the expression of CD69 on CD4+ Tregs and Th17 cells, as well as the posttraumatic expansion of platelets and hemostatic function. Subgroup analysis was performed to assess the differences between polytrauma patients with and without severe traumatic brain injury (TBI). Methods: In this non-interventional prospective clinical trial, we analyzed sequential blood samples over a period of 10 days from 30 patients after multiple traumas with an ISS ≥ 16. Platelet function was assessed by rotational thromboelastometry (ROTEM analysis). CD4+ Tregs and Th17 cells were stained with surface markers and analyzed by flow cytometry. Results: We were able to demonstrate a significantly increased expression of CD69 on CD4+ Tregs after trauma. Subgroup analysis revealed that the absence of severe TBI is associated with a significantly higher expression of CD69 on CD4+ Tregs and on Th17 cells. Platelets expanded and showed signs of dysfunction, while an overall tendency of posttraumatic hypercoagulation was detected. Conclusions: Our results support the concept of injury-specific immune responses and add to a further understanding of the complex pathophysiology of post-traumatic immune dysfunction.
The First Italian Telemedicine Program for Non-Critical COVID-19 Patients: Experience from Lodi (Italy)
Authors: Forlani Sara , Mastrosimone Erminia , Paglia Stefano , Protti Simonetta , Ferraris Maria Paola , Casale Maria Chiara , Di Capua Mirko , Grossi Maria Giuseppina , Esposti Marco , Randazzo Davide , Giustivi Davide
Published: September 2022

Italy was the first Western country to face a massive SARS-CoV-2 outbreak. The limited information initially available on the natural course of the disease required caution in the discharge of patients accessing health facilities. This resulted in overcrowded health facilities and emergency services. In this context, improvements of other forms of hospital care assistance were needed. This study shows the results of the first Italian remote monitoring program for COVID-19 patients. The program was implemented by the Azienda Socio Sanitaria (ASST) Lodi (Italy) by using the innovative Zcare software®. Data generated during patient recruitment, monitoring, and discharge were extracted from the Zcare software and statistically analysed. Data refer to a sample of 1196 patients enrolled in the remote monitoring program in 2020. Patients reported symptoms mainly during the first week. The most frequently reported symptoms were general fatigue, cough, and loss of taste and smell (dysosmia). More than 80% of patients reported a saturation level below 96% at least once, and more than 70% had a temperature above 37 °C. Active monitoring of reported symptoms provided valuable insights into the disease’s natural history during its less severe acute phase. Only 109 individuals visited the emergency department at least once in the first 100 days of monitoring. Of these, 101 had COVID-19 infection, 69 of whom were hospitalized following a first clinical assessment at the emergency department. The ASST Lodi’s telemedicine strategy for COVID patients appears to be a viable alternative to hospitalization. This strategy enables the provision of proper care while making resources available for more critically ill patients, and enhances the availability of resources available for more critically ill patients.
Comprehensive Approaches to Aspiration Pneumonia and Dysphagia in the Elderly on the Disease Time-Axis
Authors: Ebihara Takae
Published: September 2022

Pneumonia in the elderly has been increasing on an annual basis. To a greater or lesser extent, aspiration is a major contributor to the development of pneumonia in the elderly. Antimicrobials alone are not sufficient for the treatment of pneumonia, and the condition may become intractable or even recur repeatedly. In addition, some patients with pneumonia may have no problems with eating, while others are unable to receive the necessary nutrition due to severe dysphagia. It has recently been found that pneumonia decreases both the muscle mass and strength of the swallowing and respiratory muscles, a condition named pneumonia-associated sarcopenia. This contributes to a pathophysiological time-axis of aspiration pneumonia and dysphagia in the elderly, in which silent aspiration leads to the development of pneumonia, and further to dysphagia, malnutrition, and low immunity. Therefore, it is recommended that the treatment and prevention of developing pneumonia should also differ according to an individual’s placement in the disease time-axis. In particular, approaches for preventing aspiration based on scientific findings are able to be implemented at home.
Mobile-Bearing Total Ankle Replacement In Vivo Kinematic Assessment: A Prospective Study Protocol
Authors: Caravelli Silvio , Bragonzoni Laura , Vocale Emanuele , Zinno Raffaele , Di Paolo Stefano , Barone Giuseppe , De Blasiis Paolo , Benedetti Maria , Zaffagnini Stefano , Mosca Massimiliano
Published: September 2022

Total ankle prosthesis as a surgical solution in the case of end-stage osteoarthritis has seen a considerable increase in the last two decades. This study protocol arises from the need to understand the in vivo kinematics of mobile-bearing, flat tibial component total ankle replacement, evaluating the real range of motion and the reciprocal relationships between the components during normal motor tasks through the use of model-based radio-stereometric analysis (MB-RSA). In addition, pre- and post-operative evaluation of walking kinematics with inertial motion sensors, proprioception through a dedicated workstation, and clinical outcomes are discussed. We expect that based on our study protocol researchers will be able to improve future prosthetic designs and validate the setup of MB-RSA, as well as to understand “how an ankle prosthesis moves” once implanted in the patient.
Impact of Sodium Zirconium Cyclosilicate Therapy Cessation in Patients with Systolic Heart Failure
Authors: Imamura Teruhiko , Narang Nikhil , Kinugawa Koichiro
Published: September 2022

Background: Sodium zirconium cyclosilicate (SZC), a newly introduced potassium binder, is indicated for treating hyperkalemia. SZC-incorporated up-titration of renin-angiotensin system inhibitors and mineralocorticoid receptor antagonists has been recommended for those with systolic heart failure, whereas SZC is often terminated following the improvement of hyperkalemia in real-world practice. We aimed to investigate the impact of SZC cessation on the recurrence of hyperkalemia. Methods: Patients with systolic heart failure, in whom SZC was discontinued following improvement in hyperkalemia, were studied and compared to those who had continued SZC. All patients were followed for one year or until August 2022. The recurrent rates of hyperkalemia were compared between the two groups. Results: A total of 30 patients (median age 83 years, 53% men, median left ventricular ejection fraction 42%) were included. The one-year cumulative incidence of recurrent hyperkalemia was 93% in the group who discontinued SZC versus 22% in those who continued SZC (p = 0.032). In the group where SZC was withdrawn, doses of renin-angiotensin system inhibitors and mineralocorticoid receptor antagonists were less up-titrated, echocardiographic evidence of reverse remodeling occurred less, and readmission due to worsening heart failure tended to be higher compared to those who remained on SZC therapy. Conclusions: SZC cessation was associated with recurrent hyperkalemia and suboptimal medical therapy optimization compared to continuation of SZC therapy.