International Journal of Molecular Sciences

12 pages, 1270 KiB

Open AccessArticle

Understanding the Behaviour of Human Cell Types under Simulated Microgravity Conditions: The Case of Erythrocytes

by Cristina Manis, Alessia Manca, Antonio Murgia, Giuseppe Uras, Pierluigi Caboni, Terenzio Congiu, Gavino Faa, Antonella Pantaleo and Giacomo Cao

Int. J. Mol. Sci. 2022, 23(12), 6876; https://doi.org/10.3390/ijms23126876 - 20 Jun 2022

Cited by 5 | Viewed by 3096

Abstract

Erythrocytes are highly specialized cells in human body, and their main function is to ensure the gas exchanges, O₂ and CO₂, within the body. The exposure to microgravity environment leads to several health risks such as those affecting red blood [...] Read more.

Erythrocytes are highly specialized cells in human body, and their main function is to ensure the gas exchanges, O₂ and CO₂, within the body. The exposure to microgravity environment leads to several health risks such as those affecting red blood cells. In this work, we investigated the changes that occur in the structure and function of red blood cells under simulated microgravity, compared to terrestrial conditions, at different time points using biochemical and biophysical techniques. Erythrocytes exposed to simulated microgravity showed morphological changes, a constant increase in reactive oxygen species (ROS), a significant reduction in total antioxidant capacity (TAC), a remarkable and constant decrease in total glutathione (GSH) concentration, and an augmentation in malondialdehyde (MDA) at increasing times. Moreover, experiments were performed to evaluate the lipid profile of erythrocyte membranes which showed an upregulation in the following membrane phosphocholines (PC): PC16:0_16:0, PC 33:5, PC18:2_18:2, PC 15:1_20:4 and SM d42:1. Thus, remarkable changes in erythrocyte cytoskeletal architecture and membrane stiffness due to oxidative damage have been found under microgravity conditions, in addition to factors that contribute to the plasticity of the red blood cells (RBCs) including shape, size, cell viscosity and membrane rigidity. This study represents our first investigation into the effects of microgravity on erythrocytes and will be followed by other experiments towards understanding the behaviour of different human cell types in microgravity. Full article

(This article belongs to the Special Issue Advances in Membrane Protein Research)

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21 pages, 3987 KiB

Open AccessArticle

Generation and Characterization of a Tumor Stromal Microenvironment and Analysis of Its Interplay with Breast Cancer Cells: An In Vitro Model to Study Breast Cancer-Associated Fibroblast Inactivation

by Veronica Romano, Maria Rosaria Ruocco, Pietro Carotenuto, Anna Barbato, Alessandro Venuta, Vittoria Acampora, Sabrina De Lella, Elena Vigliar, Antonino Iaccarino, Giancarlo Troncone, Gaetano Calì, Luigi Insabato, Daniela Russo, Brunella Franco, Stefania Masone, Nunzio Velotti, Antonello Accurso, Tommaso Pellegrino, Giuseppe Fiume, Immacolata Belviso, Stefania Montagnani, Angelica Avagliano and Alessandro Arcucci Show full author list Hide full author list

Int. J. Mol. Sci. 2022, 23(12), 6875; https://doi.org/10.3390/ijms23126875 - 20 Jun 2022

Cited by 5 | Viewed by 3260

Abstract

Breast cancer-associated fibroblasts (BCAFs), the most abundant non-cancer stromal cells of the breast tumor microenvironment (TME), dramatically sustain breast cancer (BC) progression by interacting with BC cells. BCAFs, as well as myofibroblasts, display an up regulation of activation and inflammation markers represented by [...] Read more.

Breast cancer-associated fibroblasts (BCAFs), the most abundant non-cancer stromal cells of the breast tumor microenvironment (TME), dramatically sustain breast cancer (BC) progression by interacting with BC cells. BCAFs, as well as myofibroblasts, display an up regulation of activation and inflammation markers represented by α-smooth muscle actin (α-SMA) and cyclooxygenase 2 (COX-2). BCAF aggregates have been identified in the peripheral blood of metastatic BC patients. We generated an in vitro stromal model consisting of human primary BCAFs grown as monolayers or 3D cell aggregates, namely spheroids and reverted BCAFs, obtained from BCAF spheroids reverted to 2D cell adhesion growth after 216 h of 3D culture. We firstly evaluated the state of activation and inflammation and the mesenchymal status of the BCAF monolayers, BCAF spheroids and reverted BCAFs. Then, we analyzed the MCF-7 cell viability and migration following treatment with conditioned media from the different BCAF cultures. After 216 h of 3D culture, the BCAFs acquired an inactivated phenotype, associated with a significant reduction in α-SMA and COX-2 protein expression. The deactivation of the BCAF spheroids at 216 h was further confirmed by the cytostatic effect exerted by their conditioned medium on MCF-7 cells. Interestingly, the reverted BCAFs also retained a less activated phenotype as indicated by α-SMA protein expression reduction. Furthermore, the reverted BCAFs exhibited a reduced pro-tumor phenotype as indicated by the anti-migratory effect exerted by their conditioned medium on MCF-7 cells. The deactivation of BCAFs without drug treatment is possible and leads to a reduced capability of BCAFs to sustain BC progression in vitro. Consequently, this study could be a starting point to develop new therapeutic strategies targeting BCAFs and their interactions with cancer cells. Full article

(This article belongs to the Special Issue Novel Therapeutic Targets for Tumor Microenvironment in Cancer)

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20 pages, 4824 KiB

Open AccessArticle

Trehalose Treatment in Zebrafish Model of Lafora Disease

by Stefania Della Vecchia, Asahi Ogi, Rosario Licitra, Francesca Abramo, Gabriele Nardi, Serena Mero, Silvia Landi, Roberta Battini, Federico Sicca, Gian Michele Ratto, Filippo Maria Santorelli and Maria Marchese

Int. J. Mol. Sci. 2022, 23(12), 6874; https://doi.org/10.3390/ijms23126874 - 20 Jun 2022

Cited by 12 | Viewed by 3120

Abstract

Mutations in the EPM2A gene encoding laforin cause Lafora disease (LD), a progressive myoclonic epilepsy characterized by drug-resistant seizures and progressive neurological impairment. To date, rodents are the only available models for studying LD; however, their use for drug screening is limited by [...] Read more.

Mutations in the EPM2A gene encoding laforin cause Lafora disease (LD), a progressive myoclonic epilepsy characterized by drug-resistant seizures and progressive neurological impairment. To date, rodents are the only available models for studying LD; however, their use for drug screening is limited by regulatory restrictions and high breeding costs. To investigate the role of laforin loss of function in early neurodevelopment, and to screen for possible new compounds for treating the disorder, we developed a zebrafish model of LD. Our results showed the epm2a^−/− zebrafish to be a faithful model of LD, exhibiting the main disease features, namely motor impairment and neuronal hyperexcitability with spontaneous seizures. The model also showed increased inflammatory response and apoptotic death, as well as an altered autophagy pathway that occurs early in development and likely contributes to the disease progression. Early administration of trehalose was found to be effective for rescuing motor impairment and neuronal hyperexcitability associated with seizures. Our study adds a new tool for investigating LD and might help to identify new treatment opportunities. Full article

(This article belongs to the Special Issue Zebrafish as a Model in Human Disease)

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24 pages, 2753 KiB

Open AccessArticle

Adaptation of Oxidative Phosphorylation Machinery Compensates for Hepatic Lipotoxicity in Early Stages of MAFLD

by Pia Fahlbusch, Aleksandra Nikolic, Sonja Hartwig, Sylvia Jacob, Ulrike Kettel, Cornelia Köllmer, Hadi Al-Hasani, Stefan Lehr, Dirk Müller-Wieland, Birgit Knebel and Jörg Kotzka

Int. J. Mol. Sci. 2022, 23(12), 6873; https://doi.org/10.3390/ijms23126873 - 20 Jun 2022

Cited by 6 | Viewed by 2725

Abstract

Alterations in mitochondrial function are an important control variable in the progression of metabolic dysfunction-associated fatty liver disease (MAFLD), while also noted by increased de novo lipogenesis (DNL) and hepatic insulin resistance. We hypothesized that the organization and function of a mitochondrial electron [...] Read more.

Alterations in mitochondrial function are an important control variable in the progression of metabolic dysfunction-associated fatty liver disease (MAFLD), while also noted by increased de novo lipogenesis (DNL) and hepatic insulin resistance. We hypothesized that the organization and function of a mitochondrial electron transport chain (ETC) in this pathologic condition is a consequence of shifted substrate availability. We addressed this question using a transgenic mouse model with increased hepatic insulin resistance and DNL due to constitutively active human SREBP-1c. The abundance of ETC complex subunits and components of key metabolic pathways are regulated in the liver of these animals. Further omics approaches combined with functional assays in isolated liver mitochondria and primary hepatocytes revealed that the SREBP-1c-forced fatty liver induced a substrate limitation for oxidative phosphorylation, inducing enhanced complex II activity. The observed increased expression of mitochondrial genes may have indicated a counteraction. In conclusion, a shift of available substrates directed toward activated DNL results in increased electron flows, mainly through complex II, to compensate for the increased energy demand of the cell. The reorganization of key compounds in energy metabolism observed in the SREBP-1c animal model might explain the initial increase in mitochondrial function observed in the early stages of human MAFLD. Full article

(This article belongs to the Special Issue Nonalcoholic Fatty Liver Disease/Metabolic Associated Fatty Liver Disease: New Insights 2.0)

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16 pages, 1558 KiB

Open AccessArticle

IgG from Adult Atopic Dermatitis (AD) Patients Induces Nonatopic Neonatal Thymic Gamma–Delta T Cells (γδT) to Acquire IL-22/IL-17 Secretion Profile with Skin-Homing Properties and Epigenetic Implications Mediated by miRNA

by Beatriz Oliveira Fagundes, Thamires Rodrigues de Sousa, Andrezza Nascimento, Lorena Abreu Fernandes, Fábio da Ressureição Sgnotto, Raquel Leão Orfali, Valéria Aoki, Alberto José da Silva Duarte, Sabri Saeed Sanabani and Jefferson Russo Victor

Int. J. Mol. Sci. 2022, 23(12), 6872; https://doi.org/10.3390/ijms23126872 - 20 Jun 2022

Cited by 10 | Viewed by 2980

Abstract

γδT cells mature in the human thymus, and mainly produce IL-17A or IFN-γ, but can also produce IL-22 and modulate a variety of immune responses. Here, we aimed to evaluate whether IgG from AD patients (AD IgG) can functionally modulate thymic nonatopic γδT [...] Read more.

γδT cells mature in the human thymus, and mainly produce IL-17A or IFN-γ, but can also produce IL-22 and modulate a variety of immune responses. Here, we aimed to evaluate whether IgG from AD patients (AD IgG) can functionally modulate thymic nonatopic γδT cells. Thymic tissues were obtained from 12 infants who had not had an atopic history. Thymocytes were cultured in mock condition, or in the presence of either AD IgG or therapeutic intravenous IgG (IVIg). Following these treatments, intracellular cytokine production, phenotype, and microRNA expression profiles were investigated. AD IgG could downregulate α4β7, upregulate CLA, and induce the production of IFN-γ, IL-17, and IL-22 in γδT cells. Although both AD IgG and IVIg could directly interact with γδT cell membranes, AD IgG could reduce γδT cell apoptosis. AD IgG could upregulate nine miRNAs compared to IVIg, and six when compared to the mock condition. In parallel, some miRNAs were downregulated. Target gene prediction and functional analysis indicated that some target genes were enriched in the negative regulation of cellular transcription. This study shows that AD IgG influences the production of IL-17 and IL-22 by intrathymic nonatopic γδT cells, and demonstrates epigenetic implications mediated by miRNAs. Full article

(This article belongs to the Special Issue Interleukin 22)

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14 pages, 2526 KiB

Open AccessArticle

Diterpenoid Caesalmin C Delays Aβ-Induced Paralysis Symptoms via the DAF-16 Pathway in Caenorhabditis elegans

by Zong-Ping Zhang, Xue Bai, Wen-Bo Cui, Xiao-Han Chen, Xu Liu, De-Juan Zhi, Zhan-Xin Zhang, Dong-Qing Fei and Dong-Sheng Wang

Int. J. Mol. Sci. 2022, 23(12), 6871; https://doi.org/10.3390/ijms23126871 - 20 Jun 2022

Cited by 5 | Viewed by 2685

Abstract

Alzheimer’s disease (AD) is the most prevalent neurodegenerative disease in the world. However, there is no effective drug to cure it. Caesalmin C is a cassane-type diterpenoid abundant in Caesalpinia bonduc (Linn.) Roxb. In this study, we investigated the effect of caesalmin C [...] Read more.

Alzheimer’s disease (AD) is the most prevalent neurodegenerative disease in the world. However, there is no effective drug to cure it. Caesalmin C is a cassane-type diterpenoid abundant in Caesalpinia bonduc (Linn.) Roxb. In this study, we investigated the effect of caesalmin C on Aβ-induced toxicity and possible mechanisms in the transgenic Caenorhabditis elegans AD model. Our results showed that caesalmin C significantly alleviated the Aβ-induced paralysis phenotype in transgenic CL4176 strain C. elegans. Caesalmin C dramatically reduced the content of Aβ monomers, oligomers, and deposited spots in AD C. elegans. In addition, mRNA levels of sod-3, gst-4, and rpt-3 were up-regulated, and mRNA levels of ace-1 were down-regulated in nematodes treated with caesalmin C. The results of the RNAi assay showed that the inhibitory effect of caesalmin C on the nematode paralysis phenotype required the DAF-16 signaling pathway, but not SKN-1 and HSF-1. Further evidence suggested that caesalmin C may also have the effect of inhibiting acetylcholinesterase (AchE) and upregulating proteasome activity. These findings suggest that caesalmin C delays the progression of AD in C. elegans via the DAF-16 signaling pathway and that it could be developed into a promising medication to treat AD. Full article

(This article belongs to the Section Molecular Pharmacology)

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13 pages, 2830 KiB

Open AccessArticle

In Vivo Inhibition of TRPC6 by SH045 Attenuates Renal Fibrosis in a New Zealand Obese (NZO) Mouse Model of Metabolic Syndrome

by Zhihuang Zheng, Yao Xu, Ute Krügel, Michael Schaefer, Tilman Grune, Bernd Nürnberg, May-Britt Köhler, Maik Gollasch, Dmitry Tsvetkov and Lajos Markó

Int. J. Mol. Sci. 2022, 23(12), 6870; https://doi.org/10.3390/ijms23126870 - 20 Jun 2022

Cited by 9 | Viewed by 3343

Abstract

Metabolic syndrome is a significant worldwide public health challenge and is inextricably linked to adverse renal and cardiovascular outcomes. The inhibition of the transient receptor potential cation channel subfamily C member 6 (TRPC6) has been found to ameliorate renal outcomes in the unilateral [...] Read more.

Metabolic syndrome is a significant worldwide public health challenge and is inextricably linked to adverse renal and cardiovascular outcomes. The inhibition of the transient receptor potential cation channel subfamily C member 6 (TRPC6) has been found to ameliorate renal outcomes in the unilateral ureteral obstruction (UUO) of accelerated renal fibrosis. Therefore, the pharmacological inhibition of TPRC6 could be a promising therapeutic intervention in the progressive tubulo-interstitial fibrosis in hypertension and metabolic syndrome. In the present study, we hypothesized that the novel selective TRPC6 inhibitor SH045 (larixyl N-methylcarbamate) ameliorates UUO-accelerated renal fibrosis in a New Zealand obese (NZO) mouse model, which is a polygenic model of metabolic syndrome. The in vivo inhibition of TRPC6 by SH045 markedly decreased the mRNA expression of pro-fibrotic markers (Col1α1, Col3α1, Col4α1, Acta2, Ccn2, Fn1) and chemokines (Cxcl1, Ccl5, Ccr2) in UUO kidneys of NZO mice compared to kidneys of vehicle-treated animals. Renal expressions of intercellular adhesion molecule 1 (ICAM-1) and α-smooth muscle actin (α-SMA) were diminished in SH045- versus vehicle-treated UUO mice. Furthermore, renal inflammatory cell infiltration (F4/80+ and CD4+) and tubulointerstitial fibrosis (Sirius red and fibronectin staining) were ameliorated in SH045-treated NZO mice. We conclude that the pharmacological inhibition of TRPC6 might be a promising antifibrotic therapeutic method to treat progressive tubulo-interstitial fibrosis in hypertension and metabolic syndrome. Full article

(This article belongs to the Special Issue Chronic Kidney Disease: Underlying Molecular Mechanisms)

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23 pages, 10101 KiB

Open AccessArticle

Transcriptomic and Metabolomic Analysis of Seedling-Stage Soybean Responses to PEG-Simulated Drought Stress

by Xiyue Wang, Shuang Song, Xin Wang, Jun Liu and Shoukun Dong

Int. J. Mol. Sci. 2022, 23(12), 6869; https://doi.org/10.3390/ijms23126869 - 20 Jun 2022

Cited by 38 | Viewed by 4343

Abstract

Soybean is an important crop grown worldwide, and drought stress seriously affects the yield and quality of soybean. Therefore, it is necessary to elucidate the molecular mechanisms underlying soybean resistance to drought stress. In this study, RNA-seq technology and ultra-performance liquid chromatography–tandem mass [...] Read more.

Soybean is an important crop grown worldwide, and drought stress seriously affects the yield and quality of soybean. Therefore, it is necessary to elucidate the molecular mechanisms underlying soybean resistance to drought stress. In this study, RNA-seq technology and ultra-performance liquid chromatography–tandem mass spectrometry were used to analyze the transcriptome and metabolome changes in soybean leaves at the seedling stage under drought stress. The results showed that there were 4790 and 3483 DEGs (differentially expressed genes) and 156 and 124 DAMs (differentially expressed metabolites), respectively, in the HN65CK vs. HN65S0 and HN44CK vs. HN44S0 comparison groups. Comprehensive analysis of transcriptomic and metabolomic data reveals metabolic regulation of seedling soybean in response to drought stress. Some candidate genes such as LOC100802571, LOC100814585, LOC100777350 and LOC100787920, LOC100800547, and LOC100785313 showed different expression trends between the two cultivars, which may cause differences in drought resistance. Secondly, a large number of flavonoids were identified, and the expression of Monohydroxy-trimethoxyflavone-O-(6″-malonyl)glucoside was upregulated between the two varieties. Finally, several key candidate genes and metabolites involved in isoflavone biosynthesis and the TCA cycle were identified, suggesting that these metabolic pathways play important roles in soybean response to drought. Our study deepens the understanding of soybean drought resistance mechanisms and provides references for soybean drought resistance breeding. Full article

(This article belongs to the Special Issue Genetic Control of Plant Drought Tolerance)

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11 pages, 1744 KiB

Open AccessArticle

Novel OPN1LW/OPN1MW Exon 3 Haplotype-Associated Splicing Defect in Patients with X-Linked Cone Dysfunction

by Katarina Stingl, Britta Baumann, Pietro De Angeli, Ajoy Vincent, Elise Héon, Monique Cordonnier, Elfriede De Baere, Salmo Raskin, Mario Teruo Sato, Naoye Shiokawa, Susanne Kohl and Bernd Wissinger

Int. J. Mol. Sci. 2022, 23(12), 6868; https://doi.org/10.3390/ijms23126868 - 20 Jun 2022

Cited by 4 | Viewed by 2654

Abstract

Certain combinations of common variants in exon 3 of OPN1LW and OPN1MW, the genes encoding the apo-protein of the long- and middle-wavelength sensitive cone photoreceptor visual pigments in humans, induce splicing defects and have been associated with dyschromatopsia and cone dysfunction syndromes. [...] Read more.

Certain combinations of common variants in exon 3 of OPN1LW and OPN1MW, the genes encoding the apo-protein of the long- and middle-wavelength sensitive cone photoreceptor visual pigments in humans, induce splicing defects and have been associated with dyschromatopsia and cone dysfunction syndromes. Here we report the identification of a novel exon 3 haplotype, G-C-G-A-T-T-G-G (referring to nucleotide variants at cDNA positions c.453, c.457, c.465, c.511, c.513, c.521, c.532, and c.538) deduced to encode a pigment with the amino acid residues L-I-V-V-A at positions p.153, p.171, p.174, p.178, and p.180, in OPN1LW or OPN1MW or both in a series of seven patients from four families with cone dysfunction. Applying minigene assays for all observed exon 3 haplotypes in the patients, we demonstrated that the novel exon 3 haplotype L-I-V-V-A induces a strong but incomplete splicing defect with 3–5% of residual correctly spliced transcripts. Minigene splicing outcomes were similar in HEK293 cells and the human retinoblastoma cell line WERI-Rb1, the latter retaining a cone photoreceptor expression profile including endogenous OPN1LW and OPN1MW gene expression. Patients carrying the novel L-I-V-V-A haplotype presented with a mild form of Blue Cone Monochromacy or Bornholm Eye Disease-like phenotype with reduced visual acuity, reduced cone electroretinography responses, red-green color vision defects, and frequently with severe myopia. Full article

(This article belongs to the Special Issue Ocular Genetics)

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18 pages, 1677 KiB

Open AccessArticle

IgG from Adult Atopic Dermatitis (AD) Patients Induces Thymic IL-22 Production and CLA Expression on CD4+ T Cells: Possible Epigenetic Implications Mediated by miRNA

by Thamires Rodrigues de Sousa, Beatriz Oliveira Fagundes, Andrezza Nascimento, Lorena Abreu Fernandes, Fábio da Ressureição Sgnotto, Raquel Leão Orfali, Valéria Aoki, Alberto José da Silva Duarte, Sabri Saeed Sanabani and Jefferson Russo Victor

Int. J. Mol. Sci. 2022, 23(12), 6867; https://doi.org/10.3390/ijms23126867 - 20 Jun 2022

Cited by 9 | Viewed by 2988

Abstract

Atopic dermatitis (AD) is a common relapsing inflammatory skin disorder characterized by immune-mediated inflammation and epidermal barrier dysfunction. The pathogenesis of AD is multifactorial and has not been fully elucidated to date. This study aimed to evaluate whether serum IgG from adult AD [...] Read more.

Atopic dermatitis (AD) is a common relapsing inflammatory skin disorder characterized by immune-mediated inflammation and epidermal barrier dysfunction. The pathogenesis of AD is multifactorial and has not been fully elucidated to date. This study aimed to evaluate whether serum IgG from adult AD patients could modulate the thymic maturation of IL-22-producing T cells and CLA+ T cells of non-atopic infants. Given that miRNAs regulate immune response genes, we evaluated whether miRNA expression is also altered in cultured thymocytes. Thymocytes were cultured with purified IgG from AD patients or control conditions (mock, Intravenous-IgG (IVIg), non-atopic IgG, or atopic non-AD IgG). Using flow cytometry analysis, we assessed the expression of CLA and intracellular levels of IL-4, IFN-γ, and IL-22 on double-positive T cells (DP T), CD4 T cells, or CD8 T cells. We also investigated the frequency of IgG isotypes and their direct interaction with the thymic T cells membrane. The miRNA profiles were evaluated by the Illumina small RNA-seq approach. MiRNA target gene prediction and enrichment analyses were performed using bioinformatics. Increased frequencies of IL-22 and CLA+ producing CD4+ T cells cultured with IgG of AD patients was seen in non-atopic infant thymocytes compared to all control conditions. No alterations were observed in the frequency of IgG isotypes among evaluated IgG pools. Evidence for a direct interaction between IgG and thymic DP T, CD4 T, and CD8 T cells is presented. The small RNA-seq analysis identified ten mature miRNAs that were modulated by AD IgG compared to mock condition (miR-181b-5p, hsa-miR-130b-3p, hsa-miR-26a-5p, hsa-miR-4497, has-miR-146a, hsa-let-7i-5p, hsa-miR-342-3p, has-miR-148a-3p, has-miR-92a and has-miR-4492). The prediction of the targetome of the seven dysregulated miRNAs between AD and mock control revealed 122 putative targets, and functional and pathway enrichment analyses were performed. Our results enhance our understanding of the mechanism by which IgG can collaborate in thymic T cells in the setting of infant AD. Full article

(This article belongs to the Special Issue Interleukin 22)

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15 pages, 3437 KiB

Open AccessArticle

Short-Term, Voluntary Exercise Affects Morpho-Functional Maturation of Adult-Generated Neurons in Rat Hippocampus

by Davide Lattanzi, David Savelli, Marica Pagliarini, Riccardo Cuppini and Patrizia Ambrogini

Int. J. Mol. Sci. 2022, 23(12), 6866; https://doi.org/10.3390/ijms23126866 - 20 Jun 2022

Cited by 10 | Viewed by 2282

Abstract

Physical exercise is a well-proven neurogenic stimulus, promoting neuronal progenitor proliferation and affecting newborn cell survival. Besides, it has beneficial effects on brain health and cognition. Previously, we found that three days of physical activity in a very precocious period of adult-generated granule [...] Read more.

Physical exercise is a well-proven neurogenic stimulus, promoting neuronal progenitor proliferation and affecting newborn cell survival. Besides, it has beneficial effects on brain health and cognition. Previously, we found that three days of physical activity in a very precocious period of adult-generated granule cell life is able to antedate the appearance of the first GABAergic synaptic contacts and increase T-type Ca²⁺ channel expression. Considering the role of GABA and Ca²⁺ in fostering neuronal maturation, in this study, we used short-term, voluntary exercise on a running wheel to investigate if it is able to induce long-term morphological and synaptic changes in newborn neurons. Using adult male rats, we found that: (i) three days of voluntary physical exercise can definitively influence the morpho-functional maturation process of newborn granule neurons when applied very early during their development; (ii) a significant percentage of new neurons show more mature morphological characteristics far from the end of exercise protocol; (iii) the long-term morphological effects result in enhanced synaptic plasticity. Present findings demonstrate that the morpho-functional changes induced by exercise on very immature adult-generated neurons are permanent, affecting the neuron maturation and integration in hippocampal circuitry. Our data contribute to underpinning the beneficial potential of physical activity on brain health, also performed for short times. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Neural Circuit Development and Regeneration 2.0)

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15 pages, 3291 KiB

Open AccessArticle

RNA Polymerase II Transcription in Pneumocystis: TFIIB from Pneumocystis carinii Can Replace the Transcriptional Functions of Fission Yeast Schizosaccharomyces pombe TFIIB In Vivo and In Vitro

by Diego A. Rojas, Fabiola Urbina, Aldo Solari and Edio Maldonado

Int. J. Mol. Sci. 2022, 23(12), 6865; https://doi.org/10.3390/ijms23126865 - 20 Jun 2022

Viewed by 1947

Abstract

The Pneumocystis genus is an opportunistic fungal pathogen that infects patients with AIDS and immunocompromised individuals. The study of this fungus has been hampered due to the inability to grow it in a (defined media/pure) culture. However, the use of modern molecular techniques [...] Read more.

The Pneumocystis genus is an opportunistic fungal pathogen that infects patients with AIDS and immunocompromised individuals. The study of this fungus has been hampered due to the inability to grow it in a (defined media/pure) culture. However, the use of modern molecular techniques and genomic analysis has helped researchers to understand its complex cell biology. The transcriptional process in the Pneumocystis genus has not been studied yet, although it is assumed that it has conventional transcriptional machinery. In this work, we have characterized the function of the RNA polymerase II (RNAPII) general transcription factor TFIIB from Pneumocystis carinii using the phylogenetically related biological model Schizosaccharomyces pombe. The results of this work show that Pneumocystis carinii TFIIB is able to replace the essential function of S. pombe TFIIB both in in vivo and in vitro assays. The S. pombe strain harboring the P carinii TFIIB grew slower than the parental wild-type S. pombe strain in complete media and in minimal media. The S. pombe cells carrying out the P. carinii TFIIB are larger than the wild-type cells, indicating that the TFIIB gene replacement confers a phenotype, most likely due to defects in transcription. P. carinii TFIIB forms very weak complexes with S. pombe TATA-binding protein on a TATA box promoter but it is able to form stable complexes in vitro when S. pombe TFIIF/RNAPII are added. P. carinii TFIIB can also replace the transcriptional function of S. pombe TFIIB in an in vitro assay. The transcription start sites (TSS) of the endogenous adh gene do not change when P. carinii TFIIB replaces S. pombe TFIIB, and neither does the TSS of the nmt1 gene, although this last gene is poorly transcribed in vivo in the presence of P. carinii TFIIB. Since transcription by RNA polymerase II in Pneumocystis is poorly understood, the results described in this study are promising and indicate that TFIIB from P. carinii can replace the transcriptional functions of S. pombe TFIIB, although the cells expressing the P. carini TFIIB show an altered phenotype. However, performing studies using a heterologous approach, like this one, could be relevant to understanding the basic molecular processes of Pneumocystis such as transcription and replication. Full article

(This article belongs to the Special Issue Molecular Mechanisms on Transcription Regulation by RNA Polymerase II (Pol II))

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18 pages, 4836 KiB

Open AccessArticle

Integrated Methylome and Transcriptome Analysis Provides Insights into the DNA Methylation Underlying the Mechanism of Cytoplasmic Male Sterility in Kenaf (Hibiscus cannabinus L.)

by Zengqiang Li, Dengjie Luo, Meiqiong Tang, Shan Cao, Jiao Pan, Wenxian Zhang, Yali Hu, Jiao Yue, Zhen Huang, Ru Li and Peng Chen

Int. J. Mol. Sci. 2022, 23(12), 6864; https://doi.org/10.3390/ijms23126864 - 20 Jun 2022

Cited by 6 | Viewed by 2206

Abstract

Cytoplasmic male sterility (CMS) is widely exploited in hybrid seed production. Kenaf is an important fiber crop with high heterosis. The molecular mechanism of kenaf CMS remains unclear, particularly in terms of DNA methylation. Here, using the anthers of a kenaf CMS line [...] Read more.

Cytoplasmic male sterility (CMS) is widely exploited in hybrid seed production. Kenaf is an important fiber crop with high heterosis. The molecular mechanism of kenaf CMS remains unclear, particularly in terms of DNA methylation. Here, using the anthers of a kenaf CMS line (P3A) and its maintainer line (P3B), comparative physiological, DNA methylation, and transcriptome analyses were performed. The results showed that P3A had considerably lower levels of IAA, ABA, photosynthetic products and ATP contents than P3B. DNA methylome analysis revealed 650 differentially methylated genes (DMGs) with 313 up- and 337 down methylated, and transcriptome analysis revealed 1788 differentially expressed genes (DEGs) with 558 up- and 1230 downregulated genes in P3A compared with P3B. Moreover, 45 genes were characterized as both DEGs and DMGs, including AUX,CYP, BGL3B, SUS6, AGL30 and MYB21. Many DEGs may be regulated by related DMGs based on methylome and transcriptome studies. These DEGs were involved in carbon metabolism, plant hormone signal transduction, the TCA cycle and the MAPK signaling pathway and were shown to be important for CMS in kenaf. These results provide new insights into the epigenetic mechanism of CMS in kenaf and other crops. Full article

(This article belongs to the Special Issue Pollen-Pistil Interaction)

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15 pages, 3198 KiB

Open AccessArticle

Roles of AGD2a in Plant Development and Microbial Interactions of Lotus japonicus

by Mingchao Huang, Mengru Yuan, Chunyu Sun, Meiru Li, Pingzhi Wu, Huawu Jiang, Guojiang Wu and Yaping Chen

Int. J. Mol. Sci. 2022, 23(12), 6863; https://doi.org/10.3390/ijms23126863 - 20 Jun 2022

Viewed by 2120

Abstract

Arabidopsis AGD2 (Aberrant Growth and Death2) and its close homolog ALD1 (AGD2-like defense response protein 1) have divergent roles in plant defense. We previously reported that modulation of salicylic acid (SA) contents by ALD1 affects numbers of nodules produced by Lotus japonicus, [...] Read more.

Arabidopsis AGD2 (Aberrant Growth and Death2) and its close homolog ALD1 (AGD2-like defense response protein 1) have divergent roles in plant defense. We previously reported that modulation of salicylic acid (SA) contents by ALD1 affects numbers of nodules produced by Lotus japonicus, but AGD2′s role in leguminous plants remains unclear. A combination of enzymatic analysis and biological characterization of genetic materials was used to study the function of AGD2 (LjAGD2a and LjAGD2b) in L. japonicus. Both LjAGD2a and LjAGD2b could complement dapD and dapE mutants of Escherichia coli and had aminotransferase activity in vitro. ljagd2 plants, with insertional mutations of LjAGD2, had delayed flowering times and reduced seed weights. In contrast, overexpression of LjAGD2a in L. japonicus induced early flowering, with increases in seed and flower sizes, but reductions in pollen fertility and seed setting rates. Additionally, ljagd2a mutation resulted in increased expression of nodulin genes and corresponding increases in infection threads and nodule numbers following inoculation with Rhizobium. Changes in expression of LjAGD2a in L. japonicus also affected endogenous SA contents and hence resistance to pathogens. Our results indicate that LjAGD2a functions as an LL-DAP aminotransferase and plays important roles in plant development. Moreover, LjAGD2a activates defense signaling via the Lys synthesis pathway, thereby participating in legume–microbe interaction. Full article

(This article belongs to the Collection Advances in Plant Cell and Organism Development)

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18 pages, 4229 KiB

Open AccessArticle

Dysregulation of the Enteric Nervous System in the Mid Colon of Complement Component 3 Knockout Mice with Constipation Phenotypes

by Yun Ju Choi, Hee Jin Song, Ji Eun Kim, Su Jin Lee, You Jeong Jin, Yu Jeong Roh, Ayun Seol, Hye Sung Kim and Dae Youn Hwang

Int. J. Mol. Sci. 2022, 23(12), 6862; https://doi.org/10.3390/ijms23126862 - 20 Jun 2022

Cited by 3 | Viewed by 2895

Abstract

Complement component 3 (C3) contributes to neurogenesis, neural migration, and synaptic elimination under normal and disease conditions of the brain, even though it has not been studied in the enteric nervous system (ENS). To determine the role of C3 in the regulatory mechanism [...] Read more.

Complement component 3 (C3) contributes to neurogenesis, neural migration, and synaptic elimination under normal and disease conditions of the brain, even though it has not been studied in the enteric nervous system (ENS). To determine the role of C3 in the regulatory mechanism of ENS during C3 deficiency-induced constipation, the changes in the markers of neuronal and interstitial cells of Cajal (ICCs), the markers for excitatory and inhibitory transmission of ENS, and expression of C3 receptors were analyzed in the mid colon of C3 knockout (KO) mice at 16 weeks of age. Prominent constipation phenotypes, including the decrease in stool parameters, changes in the histological structure, and suppression of mucin secretion, were detected in C3 KO mice compared to wildtype (WT) mice. The expression levels of the neuron specific enolase (NSE), protein gene product 9.5 (PGP9.5), and C-kit markers for myenteric neurons and ICCs were lower in the mid colon of C3 KO mice than WT mice. Excitatory transmission analysis revealed similar suppression of the 5-hydroxytryptamine (5-HT) concentration, expression of 5-HT receptors, acetylcholine (ACh) concentration, ACh esterase (AChE) activity, and expression of muscarinic ACh receptors (mAChRs), despite the mAChRs downstream signaling pathway being activated in the mid colon of C3 KO mice. In inhibitory transmission analysis, C3 KO mice showed an increase in the nitric oxide (NO) concentration and inducible nitric oxide synthase (iNOS) expression, while neuronal NOS (nNOS) expression, cholecystokinin (CCK), and gastrin concentration were decreased in the same mice. Furthermore, the levels of C3a receptor (C3aR) and C3bR expression were enhanced in the mid colon of C3 KO mice compared to the WT mice during C3 deficiency-induced constipation. Overall, these results indicate that a dysregulation of the ENS may play an important role in C3 deficiency-induced constipation in the mid colon of C3 KO mice. Full article

(This article belongs to the Section Molecular Neurobiology)

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13 pages, 4334 KiB

Open AccessArticle

Genome-Wide Identification of the Salvia miltiorrhiza SmCIPK Gene Family and Revealing the Salt Resistance Characteristic of SmCIPK13

by Shuang Wang and Qi Li

Int. J. Mol. Sci. 2022, 23(12), 6861; https://doi.org/10.3390/ijms23126861 - 20 Jun 2022

Cited by 8 | Viewed by 2455

Abstract

Members of the CIPK (CBL-interacting protein kinases) gene family play important roles in calcium (Ca²⁺) signaling pathway-regulated plant resistance to abiotic stresses. Salvia miltiorrhiza, which is widely planted and grown in complex and diverse environments, is mainly focused on the [...] Read more.

Members of the CIPK (CBL-interacting protein kinases) gene family play important roles in calcium (Ca²⁺) signaling pathway-regulated plant resistance to abiotic stresses. Salvia miltiorrhiza, which is widely planted and grown in complex and diverse environments, is mainly focused on the transcriptional regulation of enzyme genes related to the biosynthesis of its bioactive components. However, the excavation of the genes related to the resistance of S.miltiorrhiza and the involved signaling pathways have not been deeply studied. In this study, 20 SmCIPK genes were identified and classified into two families and five subfamilies by biochemical means. Sequence characteristics and conserved motif analysis revealed the conservation and difference of SmCIPK protein in plants. Expression pattern analysis showed that SmCIPKs were mainly expressed in flowers and roots, and more than 90% of gene expression was induced by SA (salicylic acid), and MeJA (methyl jasmonate). Furthermore, the expression level of SmCIPK13 could be significantly increased after stress treatment with NaCl. SmCIPK13 expression in yeast reduces sensitivity to salt, while overexpression of it in Arabidopsis has the same effect and was localized in the cytoplasm, cell membrane and nucleus. In conclusion, the identification of the SmCIPK gene family and the functional characterization of the SmCIPK13 gene provides the basis for clarification of key genes in the Ca²⁺ signaling pathway and abiotic stress in S.miltiorrhiza. Full article

(This article belongs to the Topic Plant Responses and Tolerance to Salinity Stress)

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15 pages, 1950 KiB

Open AccessArticle

A Dual-Acting Nitric Oxide Donor and Phosphodiesterase 5 Inhibitor Activates Autophagy in Primary Skin Fibroblasts

by Esther Martínez-Martínez, Paola Atzei, Christine Vionnet, Carole Roubaty, Stephanie Kaeser-Pebernard, Reto Naef and Jörn Dengjel

Int. J. Mol. Sci. 2022, 23(12), 6860; https://doi.org/10.3390/ijms23126860 - 20 Jun 2022

Cited by 3 | Viewed by 2740

Abstract

Wound healing pathologies are an increasing problem in ageing societies. Chronic, non-healing wounds, which cause high morbidity and severely reduce the quality of life of affected individuals, are frequently observed in aged individuals and people suffering from diseases affected by the Western lifestyle, [...] Read more.

Wound healing pathologies are an increasing problem in ageing societies. Chronic, non-healing wounds, which cause high morbidity and severely reduce the quality of life of affected individuals, are frequently observed in aged individuals and people suffering from diseases affected by the Western lifestyle, such as diabetes. Causal treatments that support proper wound healing are still scarce. Here, we performed expression proteomics to study the effects of the small molecule TOP-N53 on primary human skin fibroblasts and keratinocytes. TOP-N53 is a dual-acting nitric oxide donor and phosphodiesterase-5 inhibitor increasing cGMP levels to support proper wound healing. In contrast to keratinocytes, which did not exhibit global proteome alterations, TOP-N53 had profound effects on the proteome of skin fibroblasts. In fibroblasts, TOP-N53 activated the cytoprotective, lysosomal degradation pathway autophagy and induced the expression of the selective autophagy receptor p62/SQSTM1. Thus, activation of autophagy might in part be responsible for beneficial effects of TOP-N53. Full article

(This article belongs to the Special Issue New Insights on Mass Spectometry Applied to Bioscience)

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14 pages, 4463 KiB

Open AccessArticle

AB186 Inhibits Migration of Triple-Negative Breast Cancer Cells and Interacts with α-Tubulin

by Marine Geoffroy, Marine Lemesle, Alexandra Kleinclauss, Sabine Mazerbourg, Levy Batista, Muriel Barberi-Heyob, Thierry Bastogne, Wilfrid Boireau, Alain Rouleau, Dorian Dupommier, Michel Boisbrun, Corinne Comoy, Stéphane Flament, Isabelle Grillier-Vuissoz and Sandra Kuntz

Int. J. Mol. Sci. 2022, 23(12), 6859; https://doi.org/10.3390/ijms23126859 - 20 Jun 2022

Cited by 4 | Viewed by 2621

Abstract

Breast cancer is one of the leading causes of cancer-related death among females worldwide. A major challenge is to develop innovative therapy in order to treat breast cancer subtypes resistant to current treatment. In the present study, we examined the effects of two [...] Read more.

Breast cancer is one of the leading causes of cancer-related death among females worldwide. A major challenge is to develop innovative therapy in order to treat breast cancer subtypes resistant to current treatment. In the present study, we examined the effects of two Troglitazone derivatives Δ2-TGZ and AB186. Previous studies showed that both compounds induce apoptosis, nevertheless AB186 was a more potent agent. The kinetic of cellular events was investigated by real-time cell analysis system (RTCA) in MCF-7 (hormone dependent) and MDA-MB-231 (triple negative) breast cancer (TNBC) cells, followed by cell morphology analysis by immuno-localization. Both compounds induced a rapid modification of both impedance-based signals and cellular morphology. This process was associated with an inhibition of cell migration measured by wound healing and transwell assays in TNBC MDA-MB-231 and Hs578T cells. In order to identify cytoplasmic targets of AB186, we performed surface plasmon resonance (SPR) and pull-down analyses. Subsequently, 6 cytoskeleton components were identified as potential targets. We further validated α-tubulin as one of the direct targets of AB186. In conclusion, our results suggested that AB186 could be promising to develop novel therapeutic strategies to treat aggressive forms of breast cancer such as TNBC. Full article

(This article belongs to the Special Issue New Drugs for Breast Cancer Treatment)

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13 pages, 3800 KiB

Open AccessArticle

Nano–Liposomes Double Loaded with Curcumin and Tetrandrine: Preparation, Characterization, Hepatotoxicity and Anti–Tumor Effects

by Jia-Wen Song, Yu-Shi Liu, Yu-Rou Guo, Wen-Xiao Zhong, Yi-Ping Guo and Li Guo

Int. J. Mol. Sci. 2022, 23(12), 6858; https://doi.org/10.3390/ijms23126858 - 20 Jun 2022

Cited by 46 | Viewed by 4308

Abstract

(1) Background: Curcumin (CUR) and tetrandrine (TET) are natural compounds with various bioactivities, but have problems with low solubility, stability, and absorption rate, resulting in low bioavailability, and limited applications in food, medicine, and other fields. It is very important to improve the [...] Read more.

(1) Background: Curcumin (CUR) and tetrandrine (TET) are natural compounds with various bioactivities, but have problems with low solubility, stability, and absorption rate, resulting in low bioavailability, and limited applications in food, medicine, and other fields. It is very important to improve the solubility while maintaining the high activity of drugs. Liposomes are micro–vesicles synthesized from cholesterol and lecithin. With high biocompatibility and biodegradability, liposomes can significantly improve drug solubility, efficacy, and bioavailability. (2) Methods: In this work, CUR and TET were encapsulated with nano–liposomes and g DSPE–MPEG 2000 (DP)was added as a stabilizer to achieve better physicochemical properties, biosafety, and anti–tumor effects. (3) Results: The nano–liposome (CT–DP–Lip) showed stable particle size (under 100 nm) under different conditions, high solubility, drug encapsulation efficiency (EE), loading capacity (LC), release rate in vitro, and stability. In addition, in vivo studies demonstrated CT–DP–Lip had no significant toxicity on zebrafish. Tumor cytotoxicity test showed that CT–DP–Lip had a strong inhibitory effect on a variety of cancer cells. (4) Conclusions: This work showed that nano–liposomes can significantly improve the physical and chemical properties of CUR and TET and make them safer and more efficient. Full article

(This article belongs to the Special Issue Nanoparticles for Tumor Imaging and Therapy)

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19 pages, 3912 KiB

Open AccessReview

Orchestrated Action of AMPK Activation and Combined VEGF/PD-1 Blockade with Lipid Metabolic Tunning as Multi-Target Therapeutics against Ovarian Cancers

by Mingo M. H. Yung, Michelle K. Y. Siu, Hextan Y. S. Ngan, David W. Chan and Karen K. L. Chan

Int. J. Mol. Sci. 2022, 23(12), 6857; https://doi.org/10.3390/ijms23126857 - 20 Jun 2022

Cited by 12 | Viewed by 5321

Abstract

Ovarian cancer is one of the most lethal gynecological malignancies worldwide, and chemoresistance is a critical obstacle in the clinical management of the disease. Recent studies have suggested that exploiting cancer cell metabolism by applying AMP-activated protein kinase (AMPK)-activating agents and distinctive adjuvant [...] Read more.

Ovarian cancer is one of the most lethal gynecological malignancies worldwide, and chemoresistance is a critical obstacle in the clinical management of the disease. Recent studies have suggested that exploiting cancer cell metabolism by applying AMP-activated protein kinase (AMPK)-activating agents and distinctive adjuvant targeted therapies can be a plausible alternative approach in cancer treatment. Therefore, the perspectives about the combination of AMPK activators together with VEGF/PD-1 blockade as a dual-targeted therapy against ovarian cancer were discussed herein. Additionally, ferroptosis, a non-apoptotic regulated cell death triggered by the availability of redox-active iron, have been proposed to be governed by multiple layers of metabolic signalings and can be synergized with immunotherapies. To this end, ferroptosis initiating therapies (FITs) and metabolic rewiring and immunotherapeutic approaches may have substantial clinical potential in combating ovarian cancer development and progression. It is hoped that the viewpoints deliberated in this review would accelerate the translation of remedial concepts into clinical trials and improve the effectiveness of ovarian cancer treatment. Full article

(This article belongs to the Special Issue Epilepsy and Metabolism)

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13 pages, 5283 KiB

Open AccessArticle

Solid-State Construction of CuO_x/Cu_1.5Mn_1.5O₄ Nanocomposite with Abundant Surface CuO_x Species and Oxygen Vacancies to Promote CO Oxidation Activity

by Baolin Liu, Hao Wu, Shihao Li, Mengjiao Xu, Yali Cao and Yizhao Li

Int. J. Mol. Sci. 2022, 23(12), 6856; https://doi.org/10.3390/ijms23126856 - 20 Jun 2022

Cited by 7 | Viewed by 2045

Abstract

Carbon monoxide (CO) oxidation performance heavily depends on the surface-active species and the oxygen vacancies of nanocomposites. Herein, the CuO_x/Cu_1.5Mn_1.5O₄ were fabricated via solid-state strategy. It is manifested that the construction of CuO_x/Cu_1.5 [...] Read more.

Carbon monoxide (CO) oxidation performance heavily depends on the surface-active species and the oxygen vacancies of nanocomposites. Herein, the CuO_x/Cu_1.5Mn_1.5O₄ were fabricated via solid-state strategy. It is manifested that the construction of CuO_x/Cu_1.5Mn_1.5O₄ nanocomposite can produce abundant surface CuO_x species and a number of oxygen vacancies, resulting in substantially enhanced CO oxidation activity. The CO is completely converted to carbon dioxide (CO₂) at 75 °C when CuO_x/Cu_1.5Mn_1.5O₄ nanocomposites were involved, which is higher than individual CuO_x, MnO_x, and Cu_1.5Mn_1.5O₄. Density function theory (DFT) calculations suggest that CO and O₂ are adsorbed on CuO_x/Cu_1.5Mn_1.5O₄ surface with relatively optimal adsorption energy, which is more beneficial for CO oxidation activity. This work presents an effective way to prepare heterogeneous metal oxides with promising application in catalysis. Full article

(This article belongs to the Special Issue Nanoparticle for Catalysis)

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20 pages, 4930 KiB

Open AccessArticle

Microtube Array Membrane Encapsulated Cell Therapy: A Novel Platform Technology Solution for Treatment of Alzheimer’s Disease

by Shu-Mei Chen, Tsung-Chin Hsu, Chee-Ho Chew, Wan-Ting Huang, Amanda Lin Chen, Yung-Feng Lin, Sabiha Eddarkaoui, Luc Buee and Chien-Chung Chen

Int. J. Mol. Sci. 2022, 23(12), 6855; https://doi.org/10.3390/ijms23126855 - 20 Jun 2022

Cited by 4 | Viewed by 3063

Abstract

Alzheimer’s disease is the most frequent form of dementia in aging population and is presently the world’s sixth largest cause of mortality. With the advancement of therapies, several solutions have been developed such as passive immunotherapy against these misfolded proteins, thereby resulting in [...] Read more.

Alzheimer’s disease is the most frequent form of dementia in aging population and is presently the world’s sixth largest cause of mortality. With the advancement of therapies, several solutions have been developed such as passive immunotherapy against these misfolded proteins, thereby resulting in the clearance. Within this segment, encapsulated cell therapy (ECT) solutions that utilize antibody releasing cells have been proposed with a multitude of techniques under development. Hence, in this study, we utilized our novel and patented Microtube Array Membranes (MTAMs) as an encapsulating platform system with anti-pTau antibody-secreting hybridoma cells to study the impact of it on Alzheimer’s disease. In vivo results revealed that in the water maze, the mice implanted with hybridoma cell MTAMs intracranially (IN) and subcutaneously (SC) showed improvement in the time spent the goal quadrant and escape latency. In passive avoidance, hybridoma cell loaded MTAMs (IN and SC) performed significantly well in step-through latency. At the end of treatment, animals with hybridoma cell loaded MTAMs had lower phosphorylated tau (pTau) expression than empty MTAMs had. Combining both experimental results unveiled that the clearance of phosphorylated tau might rescue the cognitive impairment associated with AD. Full article

(This article belongs to the Special Issue Advances in Molecular Activity of Potential Drugs 2.0)

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5 pages, 213 KiB

Open AccessEditorial

Bacterial Regulatory Proteins

by Jan Kormanec

Int. J. Mol. Sci. 2022, 23(12), 6854; https://doi.org/10.3390/ijms23126854 - 20 Jun 2022

Cited by 1 | Viewed by 1816

Abstract

The regulation of gene expression in bacteria occurs predominantly at the level of transcription, which is controlled by RNA polymerase [...] Full article

(This article belongs to the Special Issue Bacterial Regulatory Proteins)

11 pages, 1000 KiB

Open AccessArticle

Nurr1 Is Not an Essential Regulator of BDNF in Mouse Cortical Neurons

by Mona Abdollahi and Margaret Fahnestock

Int. J. Mol. Sci. 2022, 23(12), 6853; https://doi.org/10.3390/ijms23126853 - 20 Jun 2022

Cited by 5 | Viewed by 2072

Abstract

Nurr1 and brain-derived neurotrophic factor (BDNF) play major roles in cognition. Nurr1 regulates BDNF in midbrain dopaminergic neurons and cerebellar granule cells. Nurr1 and BDNF are also highly expressed in the cerebral cortex, a brain area important in cognition. Due to Nurr1 and [...] Read more.

Nurr1 and brain-derived neurotrophic factor (BDNF) play major roles in cognition. Nurr1 regulates BDNF in midbrain dopaminergic neurons and cerebellar granule cells. Nurr1 and BDNF are also highly expressed in the cerebral cortex, a brain area important in cognition. Due to Nurr1 and BDNF tissue specificity, the regulatory effect of Nurr1 on BDNF in different brain areas cannot be generalized. The relationship between Nurr1 and BDNF in the cortex has not been investigated previously. Therefore, we examined Nurr1-mediated BDNF regulation in cortical neurons in activity-dependent and activity-independent states. Mouse primary cortical neurons were treated with the Nurr1 agonist, amodiaquine (AQ). Membrane depolarization was induced by KCl or veratridine and reversed by nimodipine. AQ and membrane depolarization significantly increased Nurr1 (p < 0.001) and BDNF (p_AQ < 0.001, p_KCl < 0.01) as assessed by real-time qRT-PCR. However, Nurr1 knockdown did not affect BDNF gene expression in resting or depolarized neurons. Accordingly, the positive correlation between Nurr1 and BDNF expression in AQ and membrane depolarization experiments does not imply co-regulation because Nurr1 knockdown did not affect BDNF gene expression in resting or depolarized cortical neurons. Therefore, in contrast to midbrain dopaminergic neurons and cerebellar granule cells, Nurr1 does not regulate BDNF in cortical neurons. Full article

(This article belongs to the Special Issue Molecules Affecting Brain Development and Nervous System)

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17 pages, 4111 KiB

Open AccessArticle

A Small Molecule That Promotes Cellular Senescence Prevents Fibrogenesis and Tumorigenesis

by Moon Kee Meang, Saesbyeol Kim, Ik-Hwan Kim, Han-Soo Kim and Byung-Soo Youn

Int. J. Mol. Sci. 2022, 23(12), 6852; https://doi.org/10.3390/ijms23126852 - 20 Jun 2022

Cited by 1 | Viewed by 3264

Abstract

Uncontrolled proliferative diseases, such as fibrosis or cancer, can be fatal. We previously found that a compound containing the chromone scaffold (CS), ONG41008, had potent antifibrogenic effects associated with EMT or cell-cycle control resembling tumorigenesis. We investigated the effects of ONG41008 on tumor [...] Read more.

Uncontrolled proliferative diseases, such as fibrosis or cancer, can be fatal. We previously found that a compound containing the chromone scaffold (CS), ONG41008, had potent antifibrogenic effects associated with EMT or cell-cycle control resembling tumorigenesis. We investigated the effects of ONG41008 on tumor cells and compared these effects with those in pathogenic myofibroblasts. Stimulation of A549 (lung carcinoma epithelial cells) or PANC1 (pancreatic ductal carcinoma cells) with ONG41008 resulted in robust cellular senescence, indicating that dysregulated cell proliferation is common to fibrotic cells and tumor cells. The senescence was followed by multinucleation, a manifestation of mitotic slippage. There was significant upregulation of expression and rapid nuclear translocation of p-TP53 and p16 in the treated cancer cells, which thereafter died after 72 h confirmed by 6 day live imaging. ONG41008 exhibited a comparable senogenic potential to that of dasatinib. Interestingly, ONG41008 was only able to activate caspase-3, 7 in comparison with quercetin and fisetin, also containing CS in PANC1. ONG41008 did not seem to be essentially toxic to normal human lung fibroblasts or primary prostate epithelial cells, suggesting ONG41008 can distinguish the intracellular microenvironment between normal cells and aged or diseased cells. This effect might occur as a result of the increased NAD/NADH ratio, because ONG41008 restored this important metabolic ratio in cancer cells. Taken together, this is the first study to demonstrate that a small molecule can arrest uncontrolled proliferation during fibrogenesis or tumorigenesis via both senogenic and senolytic potential. ONG41008 could be a potential drug for a broad range of fibrotic or tumorigenic diseases. Full article

(This article belongs to the Special Issue Inflammation and Cancer 2021)

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17 pages, 1667 KiB

Open AccessReview

Serine Hydrolases in Lipid Homeostasis of the Placenta-Targets for Placental Function?

by Natascha Berger, Hanna Allerkamp and Christian Wadsack

Int. J. Mol. Sci. 2022, 23(12), 6851; https://doi.org/10.3390/ijms23126851 - 20 Jun 2022

Cited by 4 | Viewed by 3310

Abstract

The metabolic state of pregnant women and their unborn children changes throughout pregnancy and adapts to the specific needs of each gestational week. These adaptions are accomplished by the actions of enzymes, which regulate the occurrence of their endogenous substrates and products in [...] Read more.

The metabolic state of pregnant women and their unborn children changes throughout pregnancy and adapts to the specific needs of each gestational week. These adaptions are accomplished by the actions of enzymes, which regulate the occurrence of their endogenous substrates and products in all three compartments: mother, placenta and the unborn. These enzymes determine bioactive lipid signaling, supply, and storage through the generation or degradation of lipids and fatty acids, respectively. This review focuses on the role of lipid-metabolizing serine hydrolases during normal pregnancy and in pregnancy-associated pathologies, such as preeclampsia, gestational diabetes mellitus, or preterm birth. The biochemical properties of each class of lipid hydrolases are presented, with special emphasis on their role in placental function or dysfunction. While, during a normal pregnancy, an appropriate tonus of bioactive lipids prevails, dysregulation and aberrant signaling occur in diseased states. A better understanding of the dynamics of serine hydrolases across gestation and their involvement in placental lipid homeostasis under physiological and pathophysiological conditions will help to identify new targets for placental function in the future. Full article

(This article belongs to the Special Issue Emerging Role of Lipids in Metabolism and Disease – 3rd Edition)

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16 pages, 2727 KiB

Open AccessArticle

Safety of Special Waveform of Transcranial Electrical Stimulation (TES): In Vivo Assessment

by Muhammad Adeel, Chun-Ching Chen, Bor-Shing Lin, Hung-Chou Chen, Jian-Chiun Liou, Yu-Ting Li and Chih-Wei Peng

Int. J. Mol. Sci. 2022, 23(12), 6850; https://doi.org/10.3390/ijms23126850 - 20 Jun 2022

Cited by 8 | Viewed by 3155

Abstract

Intermittent theta burst (iTBS) powered by direct current stimulation (DCS) can safely be applied transcranially to induce neuroplasticity in the human and animal brain cortex. tDCS-iTBS is a special waveform that is used by very few studies, and its safety needs to be [...] Read more.

Intermittent theta burst (iTBS) powered by direct current stimulation (DCS) can safely be applied transcranially to induce neuroplasticity in the human and animal brain cortex. tDCS-iTBS is a special waveform that is used by very few studies, and its safety needs to be confirmed. Therefore, we aimed to evaluate the safety of tDCS-iTBS in an animal model after brain stimulations for 1 h and 4 weeks. Thirty-one Sprague Dawley rats were divided into two groups: (1) short-term stimulation for 1 h/session (sham, low, and high) and (2) long-term for 30 min, 3 sessions/week for 4 weeks (sham and high). The anodal stimulation applied over the primary motor cortex ranged from 2.5 to 4.5 mA/cm². The brain biomarkers and scalp tissues were assessed using ELISA and histological analysis (H&E staining) after stimulations. The caspase-3 activity, cortical myelin basic protein (MBP) expression, and cortical interleukin (IL-6) levels increased slightly in both groups compared to sham. The serum MBP, cortical neuron-specific enolase (NSE), and serum IL-6 slightly changed from sham after stimulations. There was no obvious edema or cell necrosis seen in cortical histology after the intervention. The short- and long-term stimulations did not induce significant adverse effects on brain and scalp tissues upon assessing biomarkers and conducting histological analysis. Full article

(This article belongs to the Topic Emerging Translational Research in Neurological and Psychiatric Diseases: From In Vitro to In Vivo Models, from Animals to Humans, from Qualitative to Quantitative Methods 2.0)

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16 pages, 2271 KiB

Open AccessArticle

Generation and Identification of the Number of Copies of Exogenous Genes and the T-DNA Insertion Site in SCN-Resistance Transformation Event ZHs1-2

by Guixiang Tang, Xuanbo Zhong, Wei Hong, Jianfei Li, Yue Shu and Lulu Liu

Int. J. Mol. Sci. 2022, 23(12), 6849; https://doi.org/10.3390/ijms23126849 - 20 Jun 2022

Cited by 4 | Viewed by 2062

Abstract

Soybean cyst nematode (SCN, Heterodera glycines Ichinohe) causes an estimated economic loss of about USD 3 billion each year in soybean (Glycine max L.) production worldwide. Overexpression of resistance genes against SCN provides a powerful approach to develop SCN resistance cultivars in [...] Read more.

Soybean cyst nematode (SCN, Heterodera glycines Ichinohe) causes an estimated economic loss of about USD 3 billion each year in soybean (Glycine max L.) production worldwide. Overexpression of resistance genes against SCN provides a powerful approach to develop SCN resistance cultivars in soybean. The clarification of molecular characterization in transformation events is a prerequisite for ecological risk assessment, food safety, and commercial release of genetically modified crops. Here, we generated transgenic events harboring the BCN (beet cyst nematode) resistance Hs1^pro−1 gene using the Agrobacterium-mediated method in soybean, evaluated their resistance to SCN infection, and clarified the molecular characterization of one of the transformation events. Five independent and stable inheritable transformation events were generated by an Agrobacterium-mediated transformation method. SCN resistance tests showed the average number of developed females per plant and female index (FI) in T4 ZHs1-1, ZHs1-2, ZHs1-3, ZHs1-4, and ZHs1-5 transformation events were significantly lower than that in the nontransgenic control. Among these, the ZHs1-2 transformation event had the lowest number of developed females per plant and FI. Southern hybridization showed the exogenous target Hs1^pro−1 gene was inserted in one copy and the Bar gene was inserted two copies in the ZHs1-2 transformation event. The exogenous T-DNA fragment was integrated in the reverse position of Chr02: 5351566–5231578 (mainly the Bar gene expression cassette) and in the forward position of Chr03: 17083358–17083400 (intact T-DNA, including Hs1^pro−1 and Bar gene expression cassette) using a whole genome sequencing method (WGS). The results of WGS method and Southern hybridization were consistent. All the functional elements of exogenous T-DNA fragments were verified by PCR using specific primer pairs in the T5 and T6 ZHs1-2 transformation events. These results demonstrated that the overexpression of Hs1^pro−1 gene enhanced SCN resistance, and provide an important reference for the biosafety assessment and the labeling detection in transformation event ZHs1-2. Full article

(This article belongs to the Collection Genetics and Molecular Breeding in Plants)

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22 pages, 8228 KiB

Open AccessArticle

Genomic Analysis of Waterpipe Smoke-Induced Lung Tumor Autophagy and Plasticity

by Rania Faouzi Zaarour, Mohak Sharda, Bilal Azakir, Goutham Hassan Venkatesh, Raefa Abou Khouzam, Ayesha Rifath, Zohra Nausheen Nizami, Fatima Abdullah, Fatin Mohammad, Hajar Karaali, Husam Nawafleh, Yehya Elsayed and Salem Chouaib

Int. J. Mol. Sci. 2022, 23(12), 6848; https://doi.org/10.3390/ijms23126848 - 20 Jun 2022

Cited by 3 | Viewed by 4188

Abstract

The role of autophagy in lung cancer cells exposed to waterpipe smoke (WPS) is not known. Because of the important role of autophagy in tumor resistance and progression, we investigated its relationship with WP smoking. We first showed that WPS activated autophagy, as [...] Read more.

The role of autophagy in lung cancer cells exposed to waterpipe smoke (WPS) is not known. Because of the important role of autophagy in tumor resistance and progression, we investigated its relationship with WP smoking. We first showed that WPS activated autophagy, as reflected by LC3 processing, in lung cancer cell lines. The autophagy response in smokers with lung adenocarcinoma, as compared to non-smokers with lung adenocarcinoma, was investigated further using the TCGA lung adenocarcinoma bulk RNA-seq dataset with the available patient metadata on smoking status. The results, based on a machine learning classification model using Random Forest, indicate that smokers have an increase in autophagy-activating genes. Comparative analysis of lung adenocarcinoma molecular signatures in affected patients with a long-term active exposure to smoke compared to non-smoker patients indicates a higher tumor mutational burden, a higher CD8+ T-cell level and a lower dysfunction level in smokers. While the expression of the checkpoint genes tested—PD-1, PD-L1, PD-L2 and CTLA-4—remains unchanged between smokers and non-smokers, B7-1, B7-2, IDO1 and CD200R1 were found to be higher in non-smokers than smokers. Because multiple factors in the tumor microenvironment dictate the success of immunotherapy, in addition to the expression of immune checkpoint genes, our analysis explains why patients who are smokers with lung adenocarcinoma respond better to immunotherapy, even though there are no relative differences in immune checkpoint genes in the two groups. Therefore, targeting autophagy in lung adenocarcinoma patients, in combination with checkpoint inhibitor-targeted therapies or chemotherapy, should be considered in smoker patients with lung adenocarcinoma. Full article

(This article belongs to the Special Issue Genetic and Epigenetic Regulations of Tumor Progression and Metastasis)

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Open AccessArticle

Control of STING Agonistic/Antagonistic Activity Using Amine-Skeleton-Based c-di-GMP Analogues

by Yuta Yanase, Genichiro Tsuji, Miki Nakamura, Norihito Shibata and Yosuke Demizu

Int. J. Mol. Sci. 2022, 23(12), 6847; https://doi.org/10.3390/ijms23126847 - 20 Jun 2022

Cited by 3 | Viewed by 2506

Abstract

Stimulator of Interferon Genes (STING) is a type of endoplasmic reticulum (ER)-membrane receptor. STING is activated by a ligand binding, which leads to an enhancement of the immune-system response. Therefore, a STING ligand can be used to regulate the immune system in therapeutic [...] Read more.

Stimulator of Interferon Genes (STING) is a type of endoplasmic reticulum (ER)-membrane receptor. STING is activated by a ligand binding, which leads to an enhancement of the immune-system response. Therefore, a STING ligand can be used to regulate the immune system in therapeutic strategies. However, the natural (or native) STING ligand, cyclic-di-nucleotide (CDN), is unsuitable for pharmaceutical use because of its susceptibility to degradation by enzymes and its low cell-membrane permeability. In this study, we designed and synthesized CDN derivatives by replacing the sugar-phosphodiester moiety, which is responsible for various problems of natural CDNs, with an amine skeleton. As a result, we identified novel STING ligands that activate or inhibit STING. The cyclic ligand 7, with a cyclic amine structure containing two guanines, was found to have agonistic activity, whereas the linear ligand 12 showed antagonistic activity. In addition, these synthetic ligands were more chemically stable than the natural ligands. Full article

(This article belongs to the Special Issue Small Molecules, Influence of Molecular Pathways 2.0)

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Int. J. Mol. Sci., Volume 23, Issue 12 (June-2 2022) – 520 articles

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